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Widowati W, Faried A, Gunanegara RF, Rahardja F, Zahiroh FH, Sutendi AF, Nindya FS, Azis R, Ekajaya RK. The Potential of Human Wharton's Jelly Mesenchymal Stem Cells Secretome Based Topical Gel for Therapeutic Application. Avicenna J Med Biotechnol 2024; 16:233-243. [PMID: 39606678 PMCID: PMC11589430 DOI: 10.18502/ajmb.v16i4.16739] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/29/2024] [Accepted: 07/01/2024] [Indexed: 11/29/2024] Open
Abstract
Background Diabetic Foot Ulcer (DFU) might be worsened by neuropathy and vascular issues. This condition can cause 14.3% fatality, stressing the need for effective wound healing therapy. Wound healing is a complex biological process, and human Wharton's Jelly Mesenchymal Stem Cells (hWJMSCs) may help manage DFU treatment issues. This research focuses on utilizing a gel carrier to deliver bioactive substances from Wharton's Jelly Mesenchymal Stem Cells secretome (hWJ-MSCs-Sec) as a possible treatment for DFU. Methods To maintain quality, hWJMSCs-Sec is thoroughly mixed with carbomer gel and freeze-dried. ELISA test is performed to determine the characterization of the gel of hWJMSCs-Sec such as Keratinocyte Growth Factor (KGF), Platelet-Derived Growth Factor (PDGF), Hepatocyte Growth Factor (HGF), Epidermal Growth Factor (EGF), and Heparin-Binding EGF-Like Growth Factor (HB-EGF). The antioxidant activity was also measured with Hydrogen peroxide (H2O2), Nitric oxide (NO), and Ferric Reducing Antioxidant Power (FRAP) assay. Proliferation assay was utilized using WST-8 and the wound healing potential was assessed via the migration cell ability of scratched-human skin fibroblast (BJ cells). Results The freeze-dried hWJ-MSCs-Sec showed higher levels of KGF, HGF, PDGF, EGF, HB-EGF, and the antioxidant activities compared to fresh hWJ-MSCs-Sec. Additionally, the gel of freeze-dried hWJ-MSCs-Sec exhibited higher levels compared to the gel of fresh hWJMSCs-Sec. This was evidenced by faster closure of scratched wounds on BJ cells treated with hWJMSCs-Sec and freeze-dried hWJ-MSCs-Sec gel. Conclusion The freeze-dried hWJ-MSCs-Sec gel exhibits superior quality compared to the non-freeze-dried hWJ-MSCs-Sec gel. This demonstrates that the freeze-drying procedure can maintain the bioactive chemicals found in hWJMSCs-Sec, potentially enhancing the efficacy of this gel in promoting cell regeneration for wound healing.
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Affiliation(s)
- Wahyu Widowati
- Faculty of Medicine, Maranatha Christian University, Bandung, 40164, Indonesia
| | - Ahmad Faried
- Department of Neurosurgery, Oncology & Stem Cell Working Group, Faculty of Medicine, Universitas Padjadjaran, Bandung 40161, Indonesia
- Dr. Hasan Sadikin Hospital, Bandung 40161, Indonesia
| | | | - Fanny Rahardja
- Faculty of Medicine, Maranatha Christian University, Bandung, 40164, Indonesia
| | - Fadhilah Haifa Zahiroh
- Biomolecular and Biomedical Research Center Bandung, Aretha Medika Utama, Bandung 40164, Indonesia
| | - Annisa Firdaus Sutendi
- Biomolecular and Biomedical Research Center Bandung, Aretha Medika Utama, Bandung 40164, Indonesia
| | - Faradhina Salfa Nindya
- Biomolecular and Biomedical Research Center Bandung, Aretha Medika Utama, Bandung 40164, Indonesia
| | - Rizal Azis
- Biomedical Engineering Department of Electrical Engineering, Faculty of Engineering, University of Indonesia, Depok, Indonesia
- Department of Translational Medical Science, Division of Cancer and Stem Cell, Biodiscovery Institute 3, The University of Nottingham, University Park, United Kingdom NG72RD
| | - Renandy Kristianlie Ekajaya
- Biology Study Program, Faculty of Mathematics and Science Education, Universitas Pendidikan Indonesia, Bandung, 40154, Indonesia
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Liu D, Xu Q, Meng X, Liu X, Liu J. Status of research on the development and regeneration of hair follicles. Int J Med Sci 2024; 21:80-94. [PMID: 38164355 PMCID: PMC10750333 DOI: 10.7150/ijms.88508] [Citation(s) in RCA: 8] [Impact Index Per Article: 8.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/26/2023] [Accepted: 10/17/2023] [Indexed: 01/03/2024] Open
Abstract
Hair loss, or alopecia, is a prevalent condition in modern society that imposes substantial mental and psychological burden on individuals. The types of hair loss, include androgenetic alopecia, alopecia areata, and telogen effluvium; of them, androgenetic alopecia is the most common condition. Traditional treatment modalities mainly involve medical options, such as minoxidil, finasteride and surgical interventions, such as hair transplantation. However, these treatments still have many limitations. Therefore, exploring the pathogenesis of hair loss, specifically focusing on the development and regeneration of hair follicles (HFs), and developing new strategies for promoting hair regrowth are essential. Some emerging therapies for hair loss have gained prominence; these therapies include low-level laser therapy, micro needling, fractional radio frequency, platelet-rich plasma, and stem cell therapy. The aforementioned therapeutic strategies appear promising for hair loss management. In this review, we investigated the mechanisms underlying HF development and regeneration. For this, we studied the structure, development, cycle, and cellular function of HFs. In addition, we analyzed the symptoms, types, and causes of hair loss as well as its current conventional treatments. Our study provides an overview of the most effective regenerative medicine-based therapies for hair loss.
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Affiliation(s)
| | | | | | - Xiaomei Liu
- Department of Toxicology, School of Public Health, Jilin University, Changchun 130021, China
| | - Jinyu Liu
- Department of Toxicology, School of Public Health, Jilin University, Changchun 130021, China
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Ye P, Gu R, Zhu H, Chen J, Han F, Nie X. SOX family transcription factors as therapeutic targets in wound healing: A comprehensive review. Int J Biol Macromol 2023; 253:127243. [PMID: 37806414 DOI: 10.1016/j.ijbiomac.2023.127243] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/07/2023] [Revised: 10/02/2023] [Accepted: 10/02/2023] [Indexed: 10/10/2023]
Abstract
The SOX family plays a vital role in determining the fate of cells and has garnered attention in the fields of cancer research and regenerative medicine. It also shows promise in the study of wound healing, as it actively participates in the healing processes of various tissues such as skin, fractures, tendons, and the cornea. However, our understanding of the mechanisms behind the SOX family's involvement in wound healing is limited compared to its role in cancer. Gaining insight into its role, distribution, interaction with other factors, and modifications in traumatized tissues could provide valuable new knowledge about wound healing. Based on current research, SOX2, SOX7, and SOX9 are the most promising members of the SOX family for future interventions in wound healing. SOX2 and SOX9 promote the renewal of cells, while SOX7 enhances the microvascular environment. The SOX family holds significant potential for advancing wound healing research. This article provides a comprehensive review of the latest research advancements and therapeutic tools related to the SOX family in wound healing, as well as the potential benefits and challenges of targeting the SOX family for wound treatment.
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Affiliation(s)
- Penghui Ye
- Key Lab of the Basic Pharmacology of the Ministry of Education & Joint International Research Laboratory of Ethnomedicine of Ministry of Education, Zunyi Medical University, Zunyi 563006, China; College of Pharmacy, Zunyi Medical University, Zunyi 563006, China
| | - Rifang Gu
- Key Lab of the Basic Pharmacology of the Ministry of Education & Joint International Research Laboratory of Ethnomedicine of Ministry of Education, Zunyi Medical University, Zunyi 563006, China; School Medical Office, Zunyi Medical University, Zunyi 563006, China
| | - Huan Zhu
- Key Lab of the Basic Pharmacology of the Ministry of Education & Joint International Research Laboratory of Ethnomedicine of Ministry of Education, Zunyi Medical University, Zunyi 563006, China; College of Pharmacy, Zunyi Medical University, Zunyi 563006, China
| | - Jitao Chen
- Key Lab of the Basic Pharmacology of the Ministry of Education & Joint International Research Laboratory of Ethnomedicine of Ministry of Education, Zunyi Medical University, Zunyi 563006, China; College of Pharmacy, Zunyi Medical University, Zunyi 563006, China
| | - Felicity Han
- Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD 4072, Australia
| | - Xuqiang Nie
- Key Lab of the Basic Pharmacology of the Ministry of Education & Joint International Research Laboratory of Ethnomedicine of Ministry of Education, Zunyi Medical University, Zunyi 563006, China; College of Pharmacy, Zunyi Medical University, Zunyi 563006, China; Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD 4072, Australia.
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Zhao J, Yang T, Zhou L, Liu J, Mao L, Jia R, Zhao F. Porous gelatin microspheres implanted with adipose mesenchymal stromal cells promote angiogenesis via protein kinase B/endothelial nitric oxide synthase signaling pathway in bladder reconstruction. Cytotherapy 2023; 25:1317-1330. [PMID: 37804283 DOI: 10.1016/j.jcyt.2023.08.005] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/24/2022] [Revised: 08/21/2023] [Accepted: 08/21/2023] [Indexed: 10/09/2023]
Abstract
BACKGROUND AIMS Cell failure and angiogenesis are the key to bladder wall regeneration. Three-dimensional (3D) culture using porous gelatin microspheres (GMs) as a vehicle promotes stem cell proliferation and improves the paracrine capacity of cells. This study aimed to evaluate the therapeutic potential of GMs constructed from adipose-derived mesenchymal stromal cells (ADSCs) (ADSC-GMs) combined with bladder acellular matrix (BAM) in tissue-engineered bladders. METHODS Isolation of ADSCs, flow cytometry, scanning electron microscopy and cell counting kit-8, β-galactosidase and enzyme-linked immunosorbent assays were performed in vitro to compare two-dimensional (2D) and 3D cultures. In the in vivo study, male Sprague-Dawley rats were randomly divided into three groups: the BAM replacement alone (BAM) group, ADSCs grown on BAM in replacement (ADSC) group and ADSC-GMs combined with BAM followed by replacement (ADSC-GM) group. Bladder function assessed by urodynamics after 12 weeks of bladder replacement, and the rats were sacrificed at 4 and 12 weeks for further experiments. RESULTS The in vitro results showed that GM culture promoted ADSC proliferation, inhibited apoptosis and delayed senescence compared with those in the 2D culture. In addition, ADSC-GMs increased the secretion of the angiogenic factors vascular endothelial growth factor, platelet-derived growth factor-BB, and basal fibroblast growth factor. In vivo experiments revealed that ADSC-GMs adhered to the BAM for longer than ADSCs. Moreover, ADSC-GMs significantly promoted the regeneration of bladder vessels and smooth muscle, thereby facilitating the recovery of bladder function. The expression of phosphorylated protein kinase B (AKT) and phosphorylated endothelial nitric oxide synthase (eNOS) was significantly greater in the ADSC-GMs group compared with the BAM and ADSCs groups. CONCLUSIONS ADSC-GMs increased retention of ADSCs on the BAM, thereby promoting the regeneration and functional recovery of the bladder tissue. ADSC-GMs promoted angiogenesis by activating the AKT/eNOS pathway.
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Affiliation(s)
- Jun Zhao
- Department of Urology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China
| | - Tianli Yang
- Department of Urology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China
| | - Liuhua Zhou
- Department of Urology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China
| | - Jingyu Liu
- Department of Urology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China
| | - Liang Mao
- Department of Urology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China
| | - Ruipeng Jia
- Department of Urology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China.
| | - Feng Zhao
- Department of Urology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China.
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Wang Y, Wu J, Chen J, Lu C, Liang J, Shan Y, Liu J, Li Q, Miao L, He M, Wang X, Zhang J, Wu Z. Mesenchymal stem cells paracrine proteins from three-dimensional dynamic culture system promoted wound healing in third-degree burn models. Bioeng Transl Med 2023; 8:e10569. [PMID: 38023693 PMCID: PMC10658564 DOI: 10.1002/btm2.10569] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/27/2023] [Revised: 05/30/2023] [Accepted: 06/10/2023] [Indexed: 12/01/2023] Open
Abstract
Recovery of skin function remains a significant clinical challenge for deep burns owing to the severe scar formation and poor appendage regeneration, and stem cell therapy has shown great potential for injured tissue regeneration. Here, a cell-free therapy system for deep burn skin was explored using mesenchymal stem cell paracrine proteins (MSC-PP) and polyethylene glycol (PEG) temperature-sensitive hydrogels. A three-dimensional (3D) dynamic culture system for MSCs' large-scale expansion was established using a porous gelatin microcarrier crosslinked with hyaluronic acid (PGM-HA), and the purified MSC-PP from culture supernatant was characterized by mass spectrometric analysis. The results showed the 3D dynamic culture system regulated MSCs cell cycle, reduced apoptosis, and decreased lactic acid content, and the MSC-PP produced in 3D group can promote cell proliferation, migration, and adhesion. The MSC-PP + PEG system maintained stable release in 28 days of observation in vitro. The in vivo therapeutic efficacy was investigated in the rabbit's third-degree burn model, and saline, PEG, MSC-PP, and MSC-PP + PEG treatments groups were set. The in vivo results showed that the MSC-PP + PEG group significantly improved wound healing, inhibited scar formation, and facilitated skin appendage regeneration. In conclusion, the MSC-PP + PEG sustained-release system provides a potentially effective treatment for deep burn skin healing.
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Affiliation(s)
- Yingwei Wang
- Department of OphthalmologyThe First Affiliated Hospital of Jinan UniversityGuangzhouChina
- Key Laboratory for Regenerative Medicine, Ministry of Education, Department of Developmental and Regenerative BiologyJinan UniversityGuangzhouChina
| | - Jiaxin Wu
- Key Laboratory for Regenerative Medicine, Ministry of Education, Department of Developmental and Regenerative BiologyJinan UniversityGuangzhouChina
| | - Jiamin Chen
- Key Laboratory for Regenerative Medicine, Ministry of Education, Department of Developmental and Regenerative BiologyJinan UniversityGuangzhouChina
| | - Cheng Lu
- Key Laboratory for Regenerative Medicine, Ministry of Education, Department of Developmental and Regenerative BiologyJinan UniversityGuangzhouChina
| | - Jinchao Liang
- Key Laboratory for Regenerative Medicine, Ministry of Education, Department of Developmental and Regenerative BiologyJinan UniversityGuangzhouChina
| | - Yingyi Shan
- Key Laboratory for Regenerative Medicine, Ministry of Education, Department of Developmental and Regenerative BiologyJinan UniversityGuangzhouChina
| | - Jie Liu
- Key Laboratory for Regenerative Medicine, Ministry of Education, Department of Developmental and Regenerative BiologyJinan UniversityGuangzhouChina
| | - Qi Li
- Key Laboratory for Regenerative Medicine, Ministry of Education, Department of Developmental and Regenerative BiologyJinan UniversityGuangzhouChina
| | - Liang Miao
- Burn plastic surgeryLonggang Central HospitalShenzhenChina
| | - Mu He
- Burn plastic surgeryLonggang Central HospitalShenzhenChina
| | - Xiaoying Wang
- Department of Biomedical EngineeringJinan UniversityGuangzhouChina
| | - Jianhua Zhang
- Special WardsThe First Affiliated Hospital of Jinan UniversityGuangzhouChina
| | - Zheng Wu
- Key Laboratory for Regenerative Medicine, Ministry of Education, Department of Developmental and Regenerative BiologyJinan UniversityGuangzhouChina
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Wang S, Jiang W, Lv S, Sun Z, Si L, Hu J, Yang Y, Qiu D, Liu X, Zhu S, Yang L, Qi L, Chi G, Wang G, Li P, Liao B. Human umbilical cord mesenchymal stem cells-derived exosomes exert anti-inflammatory effects on osteoarthritis chondrocytes. Aging (Albany NY) 2023; 15:9544-9560. [PMID: 37724890 PMCID: PMC10564422 DOI: 10.18632/aging.205034] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/26/2023] [Accepted: 08/25/2023] [Indexed: 09/21/2023]
Abstract
Inflammation of chondrocytes plays a critical role in the occurrence and development of osteoarthritis (OA). Recent evidence indicated exosomes derived from mesenchymal stem cells (MSCs-Exos) exhibit excellent anti-inflammatory ability in many troublesome inflammatory diseases including OA. In the present study, we aimed to explore the role of human umbilical cord-derived MSCs-Exos (hUC-MSCs-Exos) in treating the inflammation of chondrocytes and its related mechanisms. Ultracentrifugation was applied to isolate hUC-MSCs-Exos from the culture supernatant of hUC-MSCs. Two OA-like in vitro inflammation models of human articular chondrocytes induced with interleukin 1β (IL-1β) and co-incubation with macrophage utilizing transwell cell culture inserts were both used to evaluate the anti-inflammatory effects of hUC-MSCs-Exos. The mRNA sequencing of chondrocytes after treatment and microRNA (miRNA) sequencing of hUC-MSCs-Exos were detected and analyzed for possible mechanism analysis. The results of the study confirmed that hUC-MSCs-Exos had a reversed effect of IL-1β on chondrocytes in the expression of collagen type II alpha 1 (COL2A1) and matrix metalloproteinase 13 (MMP13). The addition of hUC-MSCs-Exos to M1 macrophages in the upper chamber showed down-regulation of IL-1β and tumor necrosis factor α (TNF-α), up-regulation of IL-10 and arginase1 (ARG1), and reversed the gene and protein expression of COL2A1 and MMP13 of the chondrocytes seeded in the lower chamber. The results of this study confirmed the anti-inflammatory effects of hUC-MSCs-Exos in the human articular chondrocytes inflammation model. hUC-MSCs-Exos may be used as a potential cell-free treatment strategy for chondrocyte inflammation in OA.
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Affiliation(s)
- Shichao Wang
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan 511518, Guangdong Province, People's Republic of China
- Obstetrics and Gynecology of Sino-Japanese Friendship Hospital of Jilin University, Changchun 130033, Jilin Province, People's Republic of China
| | - Wenyue Jiang
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan 511518, Guangdong Province, People's Republic of China
| | - Shuang Lv
- The Key Laboratory of Pathobiology, Ministry of Education, Department of Pathology, College of Basic Medical Sciences, Jilin University, Changchun 130021, Jilin Province, People's Republic of China
| | - Zhicheng Sun
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan 511518, Guangdong Province, People's Republic of China
| | - Lihui Si
- The Department of Obstetrics and Gynecology, Second Hospital of Jilin University, Changchun 130041, Jilin, People’s Republic of China
| | - Jinxin Hu
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan 511518, Guangdong Province, People's Republic of China
| | - Yang Yang
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan 511518, Guangdong Province, People's Republic of China
| | - Dingbang Qiu
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan 511518, Guangdong Province, People's Republic of China
| | - Xiaobin Liu
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan 511518, Guangdong Province, People's Republic of China
| | - Siying Zhu
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan 511518, Guangdong Province, People's Republic of China
| | - Lunhao Yang
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan 511518, Guangdong Province, People's Republic of China
| | - Ling Qi
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan 511518, Guangdong Province, People's Republic of China
| | - Guangfan Chi
- The Key Laboratory of Pathobiology, Ministry of Education, Department of Pathology, College of Basic Medical Sciences, Jilin University, Changchun 130021, Jilin Province, People's Republic of China
| | - Guiqing Wang
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan 511518, Guangdong Province, People's Republic of China
| | - Pengdong Li
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan 511518, Guangdong Province, People's Republic of China
- The Key Laboratory of Pathobiology, Ministry of Education, Department of Pathology, College of Basic Medical Sciences, Jilin University, Changchun 130021, Jilin Province, People's Republic of China
| | - Baojian Liao
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan 511518, Guangdong Province, People's Republic of China
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Couto PS, Stibbs DJ, Rotondi MC, Takeuchi Y, Rafiq QA. Scalable manufacturing of gene-modified human mesenchymal stromal cells with microcarriers in spinner flasks. Appl Microbiol Biotechnol 2023; 107:5669-5685. [PMID: 37470820 PMCID: PMC10439856 DOI: 10.1007/s00253-023-12634-w] [Citation(s) in RCA: 5] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/06/2023] [Revised: 06/06/2023] [Accepted: 06/11/2023] [Indexed: 07/21/2023]
Abstract
Due to their immunomodulatory properties and in vitro differentiation ability, human mesenchymal stromal cells (hMSCs) have been investigated in more than 1000 clinical trials over the last decade. Multiple studies that have explored the development of gene-modified hMSC-based products are now reaching early stages of clinical trial programmes. From an engineering perspective, the challenge lies in developing manufacturing methods capable of producing sufficient doses of ex vivo gene-modified hMSCs for clinical applications. This work demonstrates, for the first time, a scalable manufacturing process using a microcarrier-bioreactor system for the expansion of gene-modified hMSCs. Upon isolation, umbilical cord tissue mesenchymal stromal cells (UCT-hMSCs) were transduced using a lentiviral vector (LV) with green fluorescent protein (GFP) or vascular endothelial growth factor (VEGF) transgenes. The cells were then seeded in 100 mL spinner flasks using Spherecol microcarriers and expanded for seven days. After six days in culture, both non-transduced and transduced cell populations attained comparable maximum cell concentrations (≈1.8 × 105 cell/mL). Analysis of the culture supernatant identified that glucose was fully depleted after day five across the cell populations. Lactate concentrations observed throughout the culture reached a maximum of 7.5 mM on day seven. Immunophenotype analysis revealed that the transduction followed by an expansion step was not responsible for the downregulation of the cell surface receptors used to identify hMSCs. The levels of CD73, CD90, and CD105 expressing cells were above 90% for the non-transduced and transduced cells. In addition, the expression of negative markers (CD11b, CD19, CD34, CD45, and HLA-DR) was also shown to be below 5%, which is aligned with the criteria established for hMSCs by the International Society for Cell and Gene Therapy (ISCT). This work provides a foundation for the scalable manufacturing of gene-modified hMSCs which will overcome a significant translational and commercial bottleneck. KEY POINTS: • hMSCs were successfully transduced by lentiviral vectors carrying two different transgenes: GFP and VEGF • Transduced hMSCs were successfully expanded on microcarriers using spinner flasks during a period of 7 days • The genetic modification step did not cause any detrimental impact on the hMSC immunophenotype characteristics.
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Affiliation(s)
- Pedro Silva Couto
- Department of Biochemical Engineering, Advanced Centre for Biochemical Engineering, University College London, Gower Street, London, WC1E 6BT UK
| | - Dale J. Stibbs
- Department of Biochemical Engineering, Advanced Centre for Biochemical Engineering, University College London, Gower Street, London, WC1E 6BT UK
| | - Marco C. Rotondi
- Department of Biochemical Engineering, Advanced Centre for Biochemical Engineering, University College London, Gower Street, London, WC1E 6BT UK
| | - Yasuhiro Takeuchi
- Division of Infection and Immunity, University College London, Gower Street, London, WC1E 6BT UK
- Biotherapeutics and Advanced Therapies, Scientific Research and Innovation, Medicines, and Healthcare Products Regulatory Agency, South Mimms, EN6 3QG UK
| | - Qasim A. Rafiq
- Department of Biochemical Engineering, Advanced Centre for Biochemical Engineering, University College London, Gower Street, London, WC1E 6BT UK
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Ma L, He X, Wu Q. The Molecular Regulatory Mechanism in Multipotency and Differentiation of Wharton's Jelly Stem Cells. Int J Mol Sci 2023; 24:12909. [PMID: 37629090 PMCID: PMC10454700 DOI: 10.3390/ijms241612909] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/23/2023] [Revised: 08/06/2023] [Accepted: 08/10/2023] [Indexed: 08/27/2023] Open
Abstract
Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) are isolated from Wharton's jelly tissue of umbilical cords. They possess the ability to differentiate into lineage cells of three germ layers. WJ-MSCs have robust proliferative ability and strong immune modulation capacity. They can be easily collected and there are no ethical problems associated with their use. Therefore, WJ-MSCs have great tissue engineering value and clinical application prospects. The identity and functions of WJ-MSCs are regulated by multiple interrelated regulatory mechanisms, including transcriptional regulation and epigenetic modifications. In this article, we summarize the latest research progress on the genetic/epigenetic regulation mechanisms and essential signaling pathways that play crucial roles in pluripotency and differentiation of WJ-MSCs.
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Affiliation(s)
| | | | - Qiang Wu
- The State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau 999078, China
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Yang H, Zhang Y, Du Z, Wu T, Yang C. Hair follicle mesenchymal stem cell exosomal lncRNA H19 inhibited NLRP3 pyroptosis to promote diabetic mouse skin wound healing. Aging (Albany NY) 2023; 15:791-809. [PMID: 36787444 PMCID: PMC9970314 DOI: 10.18632/aging.204513] [Citation(s) in RCA: 20] [Impact Index Per Article: 10.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/30/2022] [Accepted: 02/02/2023] [Indexed: 02/16/2023]
Abstract
Skin wounds caused by diabetes are a major medical problem. Mesenchymal stem cell-derived exosomes hold promise to quicken wound healing due to their ability to transfer certain molecules to target cells, including mRNAs, microRNAs, lncRNAs, and proteins. Nonetheless, the specific mechanisms underlying this impact are not elucidated. Therefore, this research aimed to investigate the effect of MSC-derived exosomes comprising long non-coding RNA (lncRNA) H19 on diabetic skin wound healing. Hair follicle mesenchymal stem cells (HF-MSCs) were effectively isolated and detected, and exosomes (Exo) were also isolated smoothly. Pretreatment with 30 mM glucose for 24 h (HG) could efficiently induce pyroptosis in HaCaT cells. Exosomal H19 enhanced HaCaT proliferation and migration and inhibited pyroptosis by reversing the stimulation of the NLRP3 inflammasome. Injection of exosomes overexpressing lncRNA H19 to diabetic skin wound promoted sustained skin wound healing, whereas sh-H19 exosomes did not have this effect. In conclusion, Exosomes overexpressing H19 promoted HaCaT proliferation, migration and suppressed pyroptosis both in vitro and in vivo. Therefore, HFMSC-derived exosomes that overexpress H19 may be included in strategies for healing diabetic skin wounds.
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Affiliation(s)
- Hongliang Yang
- Department of Cardiology, China-Japan Union Hospital of Jilin University, Changchun 130031, China
| | - Yan Zhang
- School of Public Health, Beihua University, Jilin 132033, China
| | - Zhenwu Du
- Department of Cardiology, China-Japan Union Hospital of Jilin University, Changchun 130031, China
| | - Tengfei Wu
- Department of Laboratory Animal Science, China Medical University, Shenyang 110122, China
| | - Chun Yang
- College of Basic Medicine, Beihua University, Jilin 132033, China
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Liu Y, Amissah OB, Huangfang X, Wang L, Dieu Habimana JD, Lv L, Ding X, Li J, Chen M, Zhu J, Mukama O, Sun Y, Li Z, Huang R. Large-scale expansion of human umbilical cord-derived mesenchymal stem cells using PLGA@PLL scaffold. BIORESOUR BIOPROCESS 2023; 10:18. [PMID: 36915643 PMCID: PMC9994782 DOI: 10.1186/s40643-023-00635-6] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/10/2022] [Accepted: 01/31/2023] [Indexed: 03/16/2023] Open
Abstract
Mesenchymal stem cells (MSCs) are highly important in biomedicine and hold great potential in clinical treatment for various diseases. In recent years, the capabilities of MSCs have been under extensive investigation for practical application. Regarding therapy, the efficacy usually depends on the amount of MSCs. Nevertheless, the yield of MSCs is still limited due to the traditional cultural methods. Herein, we proposed a three-dimensional (3D) scaffold prepared using poly lactic-co-glycolic acid (PLGA) nanofiber with polylysine (PLL) grafting, to promote the growth and proliferation of MSCs derived from the human umbilical cord (hUC-MSCs). We found that the inoculated hUC-MSCs adhered efficiently to the PLGA scaffold with good affinity, fast growth rate, and good multipotency. The harvested cells were ideally distributed on the scaffold and we were able to gain a larger yield than the traditional culturing methods under the same condition. Thus, our cell seeding with a 3D scaffold could serve as a promising strategy for cell proliferation in the large-scale production of MSCs. Moreover, the simplicity and low preparation cost allow this 3D scaffold to extend its potential application beyond cell culture. Graphical Abstract Supplementary Information The online version contains supplementary material available at 10.1186/s40643-023-00635-6.
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Affiliation(s)
- Yujie Liu
- CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530 China.,Guangzhou Junyuankang Biotechnology Co., Ltd., Guangzhou, 510530 China
| | - Obed Boadi Amissah
- CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530 China.,University of Chinese Academy of Sciences, Beijing, 100049 China
| | | | - Ling Wang
- School of Life Sciences, University of Science and Technology of China, Hefei, 230027 China
| | - Jean de Dieu Habimana
- CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530 China.,University of Chinese Academy of Sciences, Beijing, 100049 China
| | - Linshuang Lv
- CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530 China.,University of Chinese Academy of Sciences, Beijing, 100049 China
| | - Xuanyan Ding
- CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530 China.,University of Chinese Academy of Sciences, Beijing, 100049 China
| | - Junyi Li
- CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530 China.,University of Chinese Academy of Sciences, Beijing, 100049 China
| | - Ming Chen
- School of Life Sciences, University of Science and Technology of China, Hefei, 230027 China
| | - Jinmin Zhu
- GZMU-GIBH Joint School of Life Sciences, Guangzhou Medical University, Guangzhou, 511436 China
| | - Omar Mukama
- CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530 China
| | - Yirong Sun
- CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530 China
| | - Zhiyuan Li
- CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530 China.,University of Chinese Academy of Sciences, Beijing, 100049 China.,School of Life Sciences, University of Science and Technology of China, Hefei, 230027 China.,GZMU-GIBH Joint School of Life Sciences, Guangzhou Medical University, Guangzhou, 511436 China.,GIBH-HKU Guangdong-Hong Kong Stem Cell and Regenerative Medicine Research Centre, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530 China.,Department of Anatomy and Neurobiology, Xiangya School of Medicine, Central South University, Changsha, 410013 China
| | - Rongqi Huang
- CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530 China.,GIBH-HKU Guangdong-Hong Kong Stem Cell and Regenerative Medicine Research Centre, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530 China
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11
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Jiao Y, Niu Y, Chen X, Luo M, Huang S, Cao T, Shi G, Wei A, Huang J. Gelatin Microspheres Loaded with Wharton's Jelly Mesenchymal Stem Cells Promote Acute Full-Thickness Skin Wound Healing and Regeneration in Mice. Adv Wound Care (New Rochelle) 2022; 12:371-386. [PMID: 36245193 DOI: 10.1089/wound.2022.0034] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022] Open
Abstract
Objective: At present, there is an urgent need to develop a novel and practical therapeutic approach to accelerate the healing of acute wounds. Mesenchymal stem cell (MSC)-based therapy is emerging as a promising therapeutic approach for acute skin wounds. However, there are still challenges in clinical application of this strategy, such as low survivability, low retention time, and less engraftment in skin wounds. Approach: Wharton's jelly mesenchymal stem cells (WJMSCs) were seeded into three-dimensional (3D) gelatin microspheres (GMs) to identify the biocompatibility of GMs. WJMSCs were embedded in GMs and then encapsulated with Pluronic F-127 (PF-127) and sodium ascorbyl phosphate (SAP) combination to transplant onto acute full-thickness skin wound in mice. Histology, immunohistochemistry, and immunofluorescence assay were used to investigate the skin wound healing, dermis regeneration, collagen deposition, cell proliferation, and neovascularization. Results: Three-dimensional GM had strong biocompatibility, compared with two-dimensional adherent culturing, GM loading increased the cell viability and proliferation ability of WJMSCs. WJMSCs+GM+PF-127+SAP transplantation increased skin wound healing rate, dermis regeneration, and type III collagen deposition through improving macrophage polarization, cell proliferation, neovascularization, cell retention, and engraftment at skin wound site. Innovation: The effective 3D encapsulation technology for WJMSCs solved the main problems of cell activity and residence time during MSC transplantation. WJMSCs+GM+PF-127+SAP transplantation will be a new and effective MSC biomaterials-based therapeutic strategy for acute skin traumatic wounds. Conclusion: WJMSCs+GM+PF-127+SAP transplantation facilitated acute full-thickness skin wound healing and regeneration and might be a new and effective therapy for acute skin traumatic wounds.
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Affiliation(s)
- Yiren Jiao
- MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China
| | - Yongxia Niu
- MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China
| | - Xiaolin Chen
- MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China
| | - Mingxun Luo
- MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China
| | - Sunxing Huang
- Key Laboratory of Reproductive Medicine of Guangdong Province, The First Affiliated Hospital and School of Life Sciences, Sun Yat-sen University, Guangzhou, China
| | - Tianqi Cao
- MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China
| | - Guang Shi
- MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China
| | - Aisheng Wei
- Foshan Hospital of Traditional Chinese Medicine, Guangzhou University of Chinese Medicine, Foshan, China
| | - Junjiu Huang
- MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China.,Key Laboratory of Reproductive Medicine of Guangdong Province, The First Affiliated Hospital and School of Life Sciences, Sun Yat-sen University, Guangzhou, China
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12
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Sharma P, Kumar A, Dey AD. Cellular Therapeutics for Chronic Wound Healing: Future for Regenerative Medicine. Curr Drug Targets 2022; 23:1489-1504. [PMID: 35748548 DOI: 10.2174/138945012309220623144620] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/20/2021] [Revised: 12/10/2021] [Accepted: 02/01/2022] [Indexed: 01/25/2023]
Abstract
Chronic wounds are associated with significant morbidity and mortality, which demand long-term effective treatment and represent a tremendous financial strain on the global healthcare systems. Regenerative medicines using stem cells have recently become apparent as a promising approach and are an active zone of investigation. They hold the potential to differentiate into specific types of cells and thus possess self-renewable, regenerative, and immune-modulatory effects. Furthermore, with the rise of technology, various cell therapies and cell types such as Bone Marrow and Adipose-derived Mesenchymal Cell (ADMSC), Endothelial Progenitor Cells (EPCs), Embryonic Stem Cells (ESCs), Mesenchymal Stem Cell (MSCs), and Pluripotent Stem Cells (PSCs) are studied for their therapeutic impact on reparative processes and tissue regeneration. Cell therapy has proven to have substantial control over enhancing the quality and rate of skin regeneration and wound restoration. The literature review brings to light the mechanics of wound healing, abnormalities resulting in chronic wounds, and the obstacles wound care researchers face, thus exploring the multitude of opportunities for potential improvement. Also, the review is focused on providing particulars on the possible cell-derived therapeutic choices and their associated challenges in healing, in the context of clinical trials, as solutions to these challenges will provide fresh and better future opportunities for improved study design and therefore yield a substantial amount of data for the development of more specialized treatments.
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Affiliation(s)
- Preety Sharma
- Chitkara College of Pharmacy, Chitkara University, Punjab, India.,Government Pharmacy College Kangra, Nagrota Bhagwan, Himachal Pradesh, India
| | - Arun Kumar
- Chitkara College of Pharmacy, Chitkara University, Punjab, India
| | - Asmita Deka Dey
- Chitkara College of Pharmacy, Chitkara University, Punjab, India
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13
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Zhang J, Li P, Zhao G, He S, Xu D, Jiang W, Peng Q, Li Z, Xie Z, Zhang H, Xu Y, Qi L. Mesenchymal stem cell-derived extracellular vesicles protect retina in a mouse model of retinitis pigmentosa by anti-inflammation through miR-146a-Nr4a3 axis. Stem Cell Res Ther 2022; 13:394. [PMID: 35922863 PMCID: PMC9351183 DOI: 10.1186/s13287-022-03100-x] [Citation(s) in RCA: 27] [Impact Index Per Article: 9.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/29/2022] [Accepted: 07/27/2022] [Indexed: 11/10/2022] Open
Abstract
Background Retinitis pigmentosa is a rod-cone degenerative disease that induces irreversible vision loss. This study probed the protective capacity of mesenchymal stem cell-derived small EVs (MSC-EVs) on the retinas of rd10 mice and the underlying mechanism.
Methods MSC-EVs were injected into the vitreous of rd10 mice at postnatal day 14 and P21; morphology and function were examined at P28. The mechanism of action was explored by using co-culture of photoreceptor cell line 661 W and microglia cell line BV2.
Results Treatment with MSC-EVs increased the survival of photoreceptors and preserved their structure. Visual function, as reflected by optomotor and electroretinogram responses, was significantly enhanced in MSC-EVs-treated rd10 mice. Mechanistically, staining for Iba1, GFAP, F4/80, CD68 and CD206 showed that MSC-EVs suppressed the activation of microglial, Müller glial and macrophages. Furthermore, western blotting showed that the treatment inhibited the NF-κB pathway. RNA-seq and qPCR showed that MSC-EVs upregulated anti-inflammatory cytokines while downregulating pro-inflammatory cytokines. MSC-EVs application in vitro decreased the number of TUNEL-positive 661 W cells co-cultured with LPS-stimulated BV2, with similar impact on the cytokine expression as in vivo study. Genetic screening predicted miR-146a to be the downstream target of MSC-EVs, which was detected in MSC-EVs and upregulated in co-cultured 661 W cells and BV2 cells after MSC-EVs treatment. Upregulation of miR-146a by using its mimic decreased the expression of the transcription factor Nr4a3, and its downregulation inhibition promoted Nr4a3 expression in both 661 W and BV2 cells. Nr4a3 was further identified as the target gene of miR-146a by dual-luciferase assay. Furthermore, overexpressing miR-146a significantly decreased the expression of LPS-induced pro-inflammatory cytokines in BV2 cells. Conclusions MSC-EVs delays retinal degeneration in rd10 mice mainly by its anti-inflammatory effect via the miR-146a-Nr4a3axis. Hence, MSC-EVs may be used in the treatment of neurodegenerative diseases. Supplementary Information The online version contains supplementary material available at 10.1186/s13287-022-03100-x.
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Affiliation(s)
- Jia Zhang
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, B24 Yinquan South Road, Qingyuan, 511518, Guangdong, China
| | - Pengdong Li
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, B24 Yinquan South Road, Qingyuan, 511518, Guangdong, China.,The Key Laboratory of Pathobiology, Department of Pathology, College of Basic Medical Sciences, Jilin University, Ministry of Education, Changchun, 130021, China
| | - Guifang Zhao
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, B24 Yinquan South Road, Qingyuan, 511518, Guangdong, China
| | - Siqi He
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, B24 Yinquan South Road, Qingyuan, 511518, Guangdong, China.,College of Basic Medicine, Beihua University, Jilin City, 132013, Jilin, China
| | - Di Xu
- Guangdong-Hong Kong-Macau Institute of Central Nervous System Regeneration, Key Laboratory of Central Nervous System Regeneration, Ministry of Education, Jinan University, Guangzhou, 510632, China
| | - Weijie Jiang
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, B24 Yinquan South Road, Qingyuan, 511518, Guangdong, China.,School of Basic Medicine, Gannan Medical University, Ganzhou, 341000, China
| | - Qian Peng
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, B24 Yinquan South Road, Qingyuan, 511518, Guangdong, China
| | - Zhaohui Li
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, B24 Yinquan South Road, Qingyuan, 511518, Guangdong, China
| | - Zhongjian Xie
- Institute of Pediatrics, Shenzhen Children's Hospital, Shenzhen, 518038, China
| | - Han Zhang
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, B24 Yinquan South Road, Qingyuan, 511518, Guangdong, China.,Shenzhen Engineering Laboratory of Phosphorene and Optoelectronics, International Collaborative Laboratory of 2D Materials for Optoelectronics Science and Technology of Ministry of Education, Shenzhen University, Shenzhen, 518060, China
| | - Ying Xu
- Guangdong-Hong Kong-Macau Institute of Central Nervous System Regeneration, Key Laboratory of Central Nervous System Regeneration, Ministry of Education, Jinan University, Guangzhou, 510632, China.
| | - Ling Qi
- The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, B24 Yinquan South Road, Qingyuan, 511518, Guangdong, China.
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14
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Wu Y, Zheng Y, Jin Z, Li S, Wu W, An C, Guo J, Zhu Z, Zhou T, Zhou Y, Cen L. Controllable manipulation of alginate-gelatin core-shell microcarriers for HUMSCs expansion. Int J Biol Macromol 2022; 216:1-13. [PMID: 35777503 DOI: 10.1016/j.ijbiomac.2022.06.173] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/25/2022] [Revised: 06/20/2022] [Accepted: 06/26/2022] [Indexed: 11/29/2022]
Abstract
Human umbilical cord mesenchymal stem cells (HUMSCs) are one of the most attractive sources of stem cells, and it is meaningful to design and develop a type of microcarriers with suitable mechanical strength for HUMSCs proliferation in order to acquire enough cells for cell-based therapy. Alginate-gelatin core-shell (AG) soft microcarriers were thus fabricated via a microfluidic device with droplet shearing/gelation facilities and surface coating for in vitro expansion of HUMSCs. The attachment and proliferation of HUMSCs on AG microcarriers with different mechanical strengths modulated by gelatin coating was studied, and the harvested cells were characterized to verity their differentiation potential. The obtained core-shell microcarriers were all uniform in size with a high mono-dispersity (CV < 5 %). An increase in the gelatin surface coating concentration from 0.5 % to 1.5 % would lead to the reduction in both the particle size of the microcarriers and swelling ratio upon the contact of culture medium, but increased elastic modulus. Microcarriers of 245.12 μm with a gelatin coating elastic modulus of 27.5 kPa (AG10) were found to be the optimal substrate for HUMSCs with an initial attachment efficiency of 44.41 % and a 5-day expansion efficiency of 647 %. The cells harvested from AG10 still reserved their outstanding pluripotency. Fresh AG10 could smoothly transfer cells from a running microcarrier-cell system of confluence to serve as a convenient way of scaling-up the existing culture. The current study thus developed suitable microcarriers, AG10, for in vitro HUMSCs expansion with well reserve of cell multipotency, and also provided a manufacturing and surface manipulating strategy of precise production and fine regulation of microcarrier properties.
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Affiliation(s)
- Yanfei Wu
- Shanghai Key Laboratory of Multiphase Materials Chemical Engineering, Department of Product Engineering, School of Chemical Engineering, East China University of Science and Technology, No.130 Mei Long Road, Shanghai 200237, China
| | - Yiling Zheng
- Shanghai Key Laboratory of Multiphase Materials Chemical Engineering, Department of Product Engineering, School of Chemical Engineering, East China University of Science and Technology, No.130 Mei Long Road, Shanghai 200237, China
| | - Ziyang Jin
- State Key Laboratory of Bioreactor Engineering, School of Bioengineering, East China University of Science and Technology, No.130 Mei Long Road, Shanghai 200237, China
| | - Shihao Li
- State Key Laboratory of Bioreactor Engineering, School of Bioengineering, East China University of Science and Technology, No.130 Mei Long Road, Shanghai 200237, China
| | - Weiqian Wu
- Shanghai Key Laboratory of Multiphase Materials Chemical Engineering, Department of Product Engineering, School of Chemical Engineering, East China University of Science and Technology, No.130 Mei Long Road, Shanghai 200237, China
| | - Chenjing An
- Shanghai Key Laboratory of Multiphase Materials Chemical Engineering, Department of Product Engineering, School of Chemical Engineering, East China University of Science and Technology, No.130 Mei Long Road, Shanghai 200237, China
| | - Jiahao Guo
- Shanghai Key Laboratory of Multiphase Materials Chemical Engineering, Department of Product Engineering, School of Chemical Engineering, East China University of Science and Technology, No.130 Mei Long Road, Shanghai 200237, China
| | - Zhihua Zhu
- Shanghai Key Laboratory of Multiphase Materials Chemical Engineering, Department of Product Engineering, School of Chemical Engineering, East China University of Science and Technology, No.130 Mei Long Road, Shanghai 200237, China
| | - Tian Zhou
- Department of Oral Maxillofacial-Head and Neck Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, College of Stomatology, Shanghai Jiao Tong University, National Center for Stomatology, National Clinical Research Center for Oral Diseases, Shanghai Key Laboratory of Stomatology, Shanghai 200011, China..
| | - Yan Zhou
- State Key Laboratory of Bioreactor Engineering, School of Bioengineering, East China University of Science and Technology, No.130 Mei Long Road, Shanghai 200237, China..
| | - Lian Cen
- Shanghai Key Laboratory of Multiphase Materials Chemical Engineering, Department of Product Engineering, School of Chemical Engineering, East China University of Science and Technology, No.130 Mei Long Road, Shanghai 200237, China.
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15
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Padhiar C, Aruni AW, Abhaya M, Muthuchamy M, Dhanraj AK, Ganesan V, Bovas FB, Rajakani SN. GMP compliant clinical grade and xenofree manufacturing of human Wharton’s jelly derived mesenchymal stem cell from pooled donors. Biochem Eng J 2022. [DOI: 10.1016/j.bej.2022.108470] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/02/2022]
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16
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17
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Egger D, Lavrentieva A, Kugelmeier P, Kasper C. Physiologic isolation and expansion of human mesenchymal stem/stromal cells for manufacturing of cell-based therapy products. Eng Life Sci 2022; 22:361-372. [PMID: 35382547 PMCID: PMC8961040 DOI: 10.1002/elsc.202100097] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/30/2021] [Revised: 10/13/2021] [Accepted: 10/15/2021] [Indexed: 01/04/2023] Open
Abstract
The utilization of mesenchymal stem/stromal cells raises new hopes in treatment of diseases and pathological conditions, while at the same time bringing immense challenges for researchers, manufacturers and physicians. It is essential to consider all steps along the in vitro fabrication of cell-based products in order to reach efficient and reproducible treatment outcomes. Here, the optimal protocols for isolation, cultivation and differentiation of mesenchymal stem cells are required. In this review we discuss these aspects and their influence on the final cell-based product quality. We demonstrate that physiological in vitro cell cultivation conditions play a crucial role in therapeutic functionalities of cultivated cells. We show that three-dimensional cell culture, dynamic culture conditions and physiologically relevant in vitro oxygen concentrations during isolation and expansion make a decisive contribution towards the improvement of cell-based products in regenerative medicine.
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Affiliation(s)
- Dominik Egger
- Department of BiotechnologyUniversity of Natural Resources and Life ScienceViennaAustria
| | | | | | - Cornelia Kasper
- Department of BiotechnologyUniversity of Natural Resources and Life ScienceViennaAustria
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18
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Panero AJ, Hirahara AM, Podesta L, Jamali AA, Andersen W, Smith AA. Allograft Tissues. ATLAS OF INTERVENTIONAL ORTHOPEDICS PROCEDURES 2022:89-101. [DOI: 10.1016/b978-0-323-75514-6.00008-x] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/03/2025]
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19
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Gorodetsky R, Aicher WK. Allogenic Use of Human Placenta-Derived Stromal Cells as a Highly Active Subtype of Mesenchymal Stromal Cells for Cell-Based Therapies. Int J Mol Sci 2021; 22:5302. [PMID: 34069909 PMCID: PMC8157571 DOI: 10.3390/ijms22105302] [Citation(s) in RCA: 24] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/27/2021] [Revised: 05/14/2021] [Accepted: 05/14/2021] [Indexed: 12/13/2022] Open
Abstract
The application of mesenchymal stromal cells (MSCs) from different sources, including bone marrow (BM, bmMSCs), adipose tissue (atMSCs), and human term placenta (hPSCs) has been proposed for various clinical purposes. Accumulated evidence suggests that the activity of the different MSCs is indirect and associated with paracrine release of pro-regenerative and anti-inflammatory factors. A major limitation of bmMSCs-based treatment for autologous application is the limited yield of cells harvested from BM and the invasiveness of the procedure. Similar effects of autologous and allogeneic MSCs isolated from various other tissues were reported. The easily available fresh human placenta seems to represent a preferred source for harvesting abundant numbers of human hPSCs for allogenic use. Cells derived from the neonate tissues of the placenta (f-hPSC) can undergo extended expansion with a low risk of senescence. The low expression of HLA class I and II on f-hPSCs reduces the risk of rejection in allogeneic or xenogeneic applications in normal immunocompetent hosts. The main advantage of hPSCs-based therapies seems to lie in the secretion of a wide range of pro-regenerative and anti-inflammatory factors. This renders hPSCs as a very competent cell for therapy in humans or animal models. This review summarizes the therapeutic potential of allogeneic applications of f-hPSCs, with reference to their indirect pro-regenerative and anti-inflammatory effects and discusses clinical feasibility studies.
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Affiliation(s)
- Raphael Gorodetsky
- Biotechnology and Radiobiology Laboratory, Sharett Institute of Oncology, Hadassah-Hebrew University Medical Center, Jerusalem 91120, Israel
| | - Wilhelm K. Aicher
- Center of Medical Research, Department of Urology at UKT, Eberhard-Karls-University, 72076 Tuebingen, Germany
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20
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Bicer M, Cottrell GS, Widera D. Impact of 3D cell culture on bone regeneration potential of mesenchymal stromal cells. Stem Cell Res Ther 2021; 12:31. [PMID: 33413646 PMCID: PMC7791873 DOI: 10.1186/s13287-020-02094-8] [Citation(s) in RCA: 25] [Impact Index Per Article: 6.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/22/2020] [Accepted: 12/10/2020] [Indexed: 12/17/2022] Open
Abstract
As populations age across the world, osteoporosis and osteoporosis-related fractures are becoming the most prevalent degenerative bone diseases. More than 75 million patients suffer from osteoporosis in the USA, the EU and Japan. Furthermore, it is anticipated that the number of patients affected by osteoporosis will increase by a third by 2050. Although conventional therapies including bisphosphonates, calcitonin and oestrogen-like drugs can be used to treat degenerative diseases of the bone, they are often associated with serious side effects including the development of oesophageal cancer, ocular inflammation, severe musculoskeletal pain and osteonecrosis of the jaw.The use of autologous mesenchymal stromal cells/mesenchymal stem cells (MSCs) is a possible alternative therapeutic approach to tackle osteoporosis while overcoming the limitations of traditional treatment options. However, osteoporosis can cause a decrease in the numbers of MSCs, induce their senescence and lower their osteogenic differentiation potential.Three-dimensional (3D) cell culture is an emerging technology that allows a more physiological expansion and differentiation of stem cells compared to cultivation on conventional flat systems.This review will discuss current understanding of the effects of different 3D cell culture systems on proliferation, viability and osteogenic differentiation, as well as on the immunomodulatory and anti-inflammatory potential of MSCs.
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Affiliation(s)
- Mesude Bicer
- Stem Cell Biology and Regenerative Medicine Group, Reading School of Pharmacy, University of Reading, PO Box 226, Whiteknights, Reading, RG6 6AP, UK
| | - Graeme S Cottrell
- Cellular and Molecular Neuroscience, School of Pharmacy, University of Reading, Reading, UK
| | - Darius Widera
- Stem Cell Biology and Regenerative Medicine Group, Reading School of Pharmacy, University of Reading, PO Box 226, Whiteknights, Reading, RG6 6AP, UK.
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21
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Silva Couto P, Rotondi M, Bersenev A, Hewitt C, Nienow A, Verter F, Rafiq Q. Expansion of human mesenchymal stem/stromal cells (hMSCs) in bioreactors using microcarriers: lessons learnt and what the future holds. Biotechnol Adv 2020; 45:107636. [DOI: 10.1016/j.biotechadv.2020.107636] [Citation(s) in RCA: 15] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/08/2020] [Revised: 08/01/2020] [Accepted: 09/22/2020] [Indexed: 02/06/2023]
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22
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Large-Scale Expansion of Human Mesenchymal Stem Cells. Stem Cells Int 2020; 2020:9529465. [PMID: 32733574 PMCID: PMC7378617 DOI: 10.1155/2020/9529465] [Citation(s) in RCA: 53] [Impact Index Per Article: 10.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/15/2020] [Revised: 06/07/2020] [Accepted: 07/01/2020] [Indexed: 12/15/2022] Open
Abstract
Mesenchymal stem cells (MSCs) are multipotent stem cells with strong immunosuppressive property that renders them an attractive source of cells for cell therapy. MSCs have been studied in multiple clinical trials to treat liver diseases, peripheral nerve damage, graft-versus-host disease, autoimmune diseases, diabetes mellitus, and cardiovascular damage. Millions to hundred millions of MSCs are required per patient depending on the disease, route of administration, frequency of administration, and patient body weight. Multiple large-scale cell expansion strategies have been described in the literature to fetch the cell quantity required for the therapy. In this review, bioprocessing strategies for large-scale expansion of MSCs were systematically reviewed and discussed. The literature search in Medline and Scopus databases identified 26 articles that met the inclusion criteria and were included in this review. These articles described the large-scale expansion of 7 different sources of MSCs using 4 different bioprocessing strategies, i.e., bioreactor, spinner flask, roller bottle, and multilayered flask. The bioreactor, spinner flask, and multilayered flask were more commonly used to upscale the MSCs compared to the roller bottle. Generally, a higher expansion ratio was achieved with the bioreactor and multilayered flask. Importantly, regardless of the bioprocessing strategies, the expanded MSCs were able to maintain its phenotype and potency. In summary, the bioreactor, spinner flask, roller bottle, and multilayered flask can be used for large-scale expansion of MSCs without compromising the cell quality.
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23
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Wyrobnik TA, Ducci A, Micheletti M. Advances in human mesenchymal stromal cell-based therapies - Towards an integrated biological and engineering approach. Stem Cell Res 2020; 47:101888. [PMID: 32688331 DOI: 10.1016/j.scr.2020.101888] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/15/2019] [Revised: 06/17/2020] [Accepted: 06/19/2020] [Indexed: 12/14/2022] Open
Abstract
Recent advances of stem cell-based therapies in clinical trials have raised the need for large-scale manufacturing platforms that can supply clinically relevant doses to meet an increasing demand. Promising results have been reported using stirred-tank bioreactors, where human Mesenchymal Stromal Cells (hMSCs) were cultured in suspension on microcarriers (MCs), although the formation of microcarrier-cell-aggregates might still limit mass transfer and determine a heterogeneous distribution of hMSCs. A variety of MCs, bioreactor-impeller configurations, and agitation conditions have been established in an attempt to overcome the trade-off of ensuring good suspension while keeping the stresses to a minimum. While understanding and controlling the fluid flow environment of bioreactors has been initially under-appreciated, it has recently gained in popularity in the mission of providing ideal culture environments across different scales. This review article aims to provide a comprehensive overview of how rigorous engineering characterisation studies improved the outcome of biological process development and scale-up efforts. Reconciling these two disciplines is crucial to propose tailored bioprocessing solutions that can provide improved growth environments across a range of scales for the allogeneic cell therapies of the future.
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Affiliation(s)
- Tom A Wyrobnik
- Department of Biochemical Engineering, UCL, Gower Street, London WC1E 6BT, UK
| | - Andrea Ducci
- Department of Mechanical Engineering, UCL, Torrington Place, London WC1E 7JE, UK
| | - Martina Micheletti
- Department of Biochemical Engineering, UCL, Gower Street, London WC1E 6BT, UK.
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24
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Zhao G, Liu F, Liu Z, Zuo K, Wang B, Zhang Y, Han X, Lian A, Wang Y, Liu M, Zou F, Li P, Liu X, Jin M, Liu JY. MSC-derived exosomes attenuate cell death through suppressing AIF nucleus translocation and enhance cutaneous wound healing. Stem Cell Res Ther 2020; 11:174. [PMID: 32393338 PMCID: PMC7212595 DOI: 10.1186/s13287-020-01616-8] [Citation(s) in RCA: 70] [Impact Index Per Article: 14.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/03/2019] [Revised: 02/02/2020] [Accepted: 02/24/2020] [Indexed: 02/06/2023] Open
Abstract
Background Skin wounding is very common and may be slow to heal. Increasing evidence shows that exosomes derived from mesenchymal stem cells (MSCs) dramatically enhance skin wound healing in a paracrine manner. However, the mechanism underlying this phenomenon has not yet been elucidated. Thus, the objective of the present study was to identify the signaling pathways and paracrine factors by which MSC-derived exosomes promote de novo skin tissue regeneration in response to wound healing. Methods In vitro and in vivo skin wound healing models were created by treating immortalized human keratinocytes (HaCaT) with hydrogen peroxide (H2O2) and excising full-thickness mouse skin, respectively. Exosomes were extracted from human umbilical cord Wharton’s jelly MSCs (hucMSC-Ex) by ultracentrifugation of cell culture supernatant. Results The hucMSC-Ex treatment significantly increased HaCaT cell proliferation and migration in a time- and dose-dependent manner, suppressed HaCaT apoptosis induced with H2O2 by inhibiting nuclear translocation of apoptosis-inducing factor (AIF) and upregulating poly ADP ribose polymerase 1 (PARP-1) and poly (ADP-ribose) (PAR). The animal experiments showed that relative to hucMSCs, hucMSC-Ex attenuated full-thickness skin wounding by enhancing epidermal re-epithelialization and dermal angiogenesis. Conclusions These findings indicated that direct administration of hucMSC-Ex may effectively treat cutaneous wounding and could be of great value in clinical settings.
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Affiliation(s)
- Guifang Zhao
- Department of Toxicology, School of Public Health, Jilin University, No. 1163 Xinmin Street, Changchun, Jilin, 130021, China.,Department of Pathology, Jilin Medical University, Jilin, China
| | - Feilin Liu
- Department of Ophthalmology, the Second Hospital of Jilin University, Changchun, China
| | - Zinan Liu
- Department of Toxicology, School of Public Health, Jilin University, No. 1163 Xinmin Street, Changchun, Jilin, 130021, China
| | - Kuiyang Zuo
- Department of Toxicology, School of Public Health, Jilin University, No. 1163 Xinmin Street, Changchun, Jilin, 130021, China
| | - Bo Wang
- Department of Toxicology, School of Public Health, Jilin University, No. 1163 Xinmin Street, Changchun, Jilin, 130021, China
| | - Yuying Zhang
- Department of Toxicology, School of Public Health, Jilin University, No. 1163 Xinmin Street, Changchun, Jilin, 130021, China
| | - Xing Han
- Department of Toxicology, School of Public Health, Jilin University, No. 1163 Xinmin Street, Changchun, Jilin, 130021, China
| | - Aobo Lian
- Department of Toxicology, School of Public Health, Jilin University, No. 1163 Xinmin Street, Changchun, Jilin, 130021, China
| | - Yuan Wang
- Department of Toxicology, School of Public Health, Jilin University, No. 1163 Xinmin Street, Changchun, Jilin, 130021, China
| | - Mingsheng Liu
- Department of Toxicology, School of Public Health, Jilin University, No. 1163 Xinmin Street, Changchun, Jilin, 130021, China
| | - Fei Zou
- Department of Toxicology, School of Public Health, Jilin University, No. 1163 Xinmin Street, Changchun, Jilin, 130021, China
| | - Pengdong Li
- Department of Toxicology, School of Public Health, Jilin University, No. 1163 Xinmin Street, Changchun, Jilin, 130021, China
| | - Xiaomei Liu
- Department of Toxicology, School of Public Health, Jilin University, No. 1163 Xinmin Street, Changchun, Jilin, 130021, China
| | - Minghua Jin
- Department of Toxicology, School of Public Health, Jilin University, No. 1163 Xinmin Street, Changchun, Jilin, 130021, China
| | - Jin Yu Liu
- Department of Toxicology, School of Public Health, Jilin University, No. 1163 Xinmin Street, Changchun, Jilin, 130021, China.
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25
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Yan X, Zhang K, Yang Y, Deng D, Lyu C, Xu H, Liu W, Du Y. Dispersible and Dissolvable Porous Microcarrier Tablets Enable Efficient Large-Scale Human Mesenchymal Stem Cell Expansion. Tissue Eng Part C Methods 2020; 26:263-275. [PMID: 32268824 DOI: 10.1089/ten.tec.2020.0039] [Citation(s) in RCA: 41] [Impact Index Per Article: 8.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/14/2022] Open
Abstract
Human mesenchymal stem cells (hMSCs) have wide applications in regenerative medicine but their clinical translation is largely hindered by limited production capacity of current cell expansion regime. This study utilizes a novel dispersible and dissolvable porous microcarrier tablet, 3D TableTrix™, in stirred bioreactor to demonstrate a scalable expansion protocol for industrial manufacturing of hMSCs. The 3D TableTrix is a ready-to-use tablet that disperses into 10s of 1000s porous microcarriers upon contact with culture media, eliminating the need to prepare microcarriers before cell seeding, hence simplifying operation process. We demonstrate over 500 times expansion of adipose-derived hMSCs using serum-free culture medium in 11 days with bead-to-bead transfer for a partial scale-up from laboratory-scale spinner flasks to a 1-L bioreactor system. A final yield of 1.05 ± 0.11 × 109 hMSCs was achieved, and yield of over 3 × 109 with an overall expansion factor of 1530 could theoretically be realized with full scale-up. Cells were harvested by dissolving microcarriers with 98.6% ± 0.1% recovery rate. Cells retained their immunophenotypic characteristics, trilineage differentiation potential, and genome stability with low indications of senescence phenotype. This study illuminates the potential of industrializing clinical-grade hMSC production using 3D TableTrix microcarrier tablets and stirred tank bioreactors. Impact statement The 3D TableTrix™ is a newly available microcarrier ingeniously designed as dispersible and dissolvable porous microcarrier tablets for human mesenchymal stem cell (hMSC) expansion. This eliminates the need of tedious preparation work usually required for microcarriers and its dissolvable nature allows for high cell recovery rate of 98.6% ± 0.1%. Over 500 times expansion of adipose-derived mesenchymal stem cells in serum-free culture media using a 1-L bioreactor system demonstrates its tremendous potential for industrial production of hMSCs.
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Affiliation(s)
- Xiaojun Yan
- Department of Biomedical Engineering, School of Medicine, Tsinghua-PKU Center for Life Sciences, Tsinghua University, Beijing, China
| | - Kun Zhang
- Department of Biomedical Engineering, School of Medicine, Tsinghua-PKU Center for Life Sciences, Tsinghua University, Beijing, China
| | - Yanping Yang
- Department of Biomedical Engineering, School of Medicine, Tsinghua-PKU Center for Life Sciences, Tsinghua University, Beijing, China
| | - Dongkai Deng
- Department of Biomedical Engineering, School of Medicine, Tsinghua-PKU Center for Life Sciences, Tsinghua University, Beijing, China
| | - Cheng Lyu
- Beijing CytoNiche Biotechnology Co. Ltd., Beijing, China
| | - Huanye Xu
- Department of Biomedical Engineering, School of Medicine, Tsinghua-PKU Center for Life Sciences, Tsinghua University, Beijing, China
| | - Wei Liu
- Department of Biomedical Engineering, School of Medicine, Tsinghua-PKU Center for Life Sciences, Tsinghua University, Beijing, China.,Beijing CytoNiche Biotechnology Co. Ltd., Beijing, China
| | - Yanan Du
- Department of Biomedical Engineering, School of Medicine, Tsinghua-PKU Center for Life Sciences, Tsinghua University, Beijing, China
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26
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He Q, Zhang J, Liao Y, Alakpa EV, Bunpetch V, Zhang J, Ouyang H. Current advances in microsphere based cell culture and tissue engineering. Biotechnol Adv 2019; 39:107459. [PMID: 31682922 DOI: 10.1016/j.biotechadv.2019.107459] [Citation(s) in RCA: 37] [Impact Index Per Article: 6.2] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/21/2019] [Revised: 09/12/2019] [Accepted: 10/01/2019] [Indexed: 12/12/2022]
Affiliation(s)
- Qiulin He
- Department of Orthopaedic Surgery, Second Affiliated Hospital & Zhejiang University-University of Edinburgh Institute & School of Basic Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.; Dr. Li Dak Sum & Yip Yio Chin Center for Stem Cells and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.; Key Laboratory of Tissue Engineering and Regenerative Medicine of Zhejiang Province, Zhejiang University School of Medicine, Hangzhou 310058, China
| | - Jingwei Zhang
- Dr. Li Dak Sum & Yip Yio Chin Center for Stem Cells and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.; Key Laboratory of Tissue Engineering and Regenerative Medicine of Zhejiang Province, Zhejiang University School of Medicine, Hangzhou 310058, China
| | - Youguo Liao
- Dr. Li Dak Sum & Yip Yio Chin Center for Stem Cells and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.; Key Laboratory of Tissue Engineering and Regenerative Medicine of Zhejiang Province, Zhejiang University School of Medicine, Hangzhou 310058, China.; Guangxi Collaborative Innovation Center for Biomedicine, Guangxi Medical University, Nanning 530021, China
| | - Enateri Verissarah Alakpa
- Department of Orthopaedic Surgery, Second Affiliated Hospital & Zhejiang University-University of Edinburgh Institute & School of Basic Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China
| | - Varitsara Bunpetch
- Dr. Li Dak Sum & Yip Yio Chin Center for Stem Cells and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.; Key Laboratory of Tissue Engineering and Regenerative Medicine of Zhejiang Province, Zhejiang University School of Medicine, Hangzhou 310058, China
| | - Jiayan Zhang
- Dr. Li Dak Sum & Yip Yio Chin Center for Stem Cells and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.; Key Laboratory of Tissue Engineering and Regenerative Medicine of Zhejiang Province, Zhejiang University School of Medicine, Hangzhou 310058, China
| | - Hongwei Ouyang
- Department of Orthopaedic Surgery, Second Affiliated Hospital & Zhejiang University-University of Edinburgh Institute & School of Basic Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.; Dr. Li Dak Sum & Yip Yio Chin Center for Stem Cells and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.; Key Laboratory of Tissue Engineering and Regenerative Medicine of Zhejiang Province, Zhejiang University School of Medicine, Hangzhou 310058, China.; Department of Sports Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.; China Orthopedic Regenerative Medicine Group (CORMed), China..
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27
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Fu Y, Kong Y, Li J, Wang Y, Li M, Wang Y, Wang Y, Ren F, Ni J, Li Y, Chang Z. Mesenchymal stem cells combined with traditional Chinese medicine (qi-fang-bi-min-tang) alleviates rodent allergic rhinitis. J Cell Biochem 2019; 121:1541-1551. [PMID: 31535402 DOI: 10.1002/jcb.29389] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/15/2018] [Accepted: 03/15/2019] [Indexed: 12/12/2022]
Abstract
Mesenchymal stem cells (MSCs) have been proved to exert anti-inflammatory effects and regulate immune reactions. Traditional Chinese medicine (TCM), qi-fang-bi-min-tang, is effective for some patients with allergic diseases. However, it remains unclear whether MSCs combined with TCM could benefit the treatment of allergic rhinitis (AR). In this study, we reported an additional effect of TCM (qi-fang-bi-min-tang) on the therapy of AR under MSCs treatment. Intriguingly, we observed that TCM-treated MSCs significantly inhibited the symptoms of AR and reduced the pathological changes of nasal mucosa in ovalbumin (OVA)-induced rats. The expression levels of interferon γ (IFN-γ), interleukin-17 (IL-17), and IL-4 were significantly decreased in the plasma of AR rats after injection of TCM-treated MSCs. TCM-treated MSCs reduced the levels of histamine secreted by mast cells and immunoglobulin E (IgE) secreted by plasma cells. In addition, we found that MSCs combined with TCM had a better therapeutic effect than TCM alone on AR in an OVA-induced mouse model. After OVA induction, MSCs combined with TCM significantly reduced the ratio of T helper type 1 (Th1), Th2, and Th17, but increased the proportion of Treg in the spleen of mice. Consistently, the expression levels of IFN-γ, IL-4, and IL-17 were significantly decreased, but transforming growth factor-β1 was significantly increased in the plasma of AR mice after treated with TCM and MSCs. Our results from both rats and mice indicated that the effects of TCM combined with MSCs on the AR might be through regulating the secretion of Th1, Th2, and Th17 cytokines. This study suggested that TCM (qi-fang-bi-min-tang)-treated MSCs could be used in the clinical therapy of AR.
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Affiliation(s)
- Yanxia Fu
- School of Medicine, State Key Laboratory of Membrane Biology, Tsinghua University, Beijing, China.,Tsinghua University-Perking University Joint Center for Life Sciences, Beijing, China
| | - Yanhua Kong
- Beijing Key Laboratory, The Key Institute of state Administration of Traditional Chinese Medicine, Beijing, China
| | - Jun Li
- Institute of Immunology, PLA, The Third Military Medical University, Chongqing, China
| | - Yi Wang
- School of Medicine, State Key Laboratory of Membrane Biology, Tsinghua University, Beijing, China
| | - Mengdi Li
- School of Medicine, State Key Laboratory of Membrane Biology, Tsinghua University, Beijing, China
| | - Ying Wang
- School of Medicine, State Key Laboratory of Membrane Biology, Tsinghua University, Beijing, China
| | - Yinyin Wang
- School of Medicine, State Key Laboratory of Membrane Biology, Tsinghua University, Beijing, China
| | - Fangli Ren
- School of Medicine, State Key Laboratory of Membrane Biology, Tsinghua University, Beijing, China
| | - Jian Ni
- Institute of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, China
| | - Youlin Li
- Beijing Key Laboratory, The Key Institute of state Administration of Traditional Chinese Medicine, Beijing, China
| | - Zhijie Chang
- School of Medicine, State Key Laboratory of Membrane Biology, Tsinghua University, Beijing, China
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28
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Chopra N, Dutt Arya B, Jain N, Yadav P, Wajid S, Singh SP, Choudhury S. Biophysical Characterization and Drug Delivery Potential of Exosomes from Human Wharton's Jelly-Derived Mesenchymal Stem Cells. ACS OMEGA 2019; 4:13143-13152. [PMID: 31460441 PMCID: PMC6705090 DOI: 10.1021/acsomega.9b01180] [Citation(s) in RCA: 36] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/24/2019] [Accepted: 07/24/2019] [Indexed: 05/22/2023]
Abstract
Cell-derived exosomes (30-200 nm) as biological "nanocarriers" have attracted a great deal of interest for therapeutic applications due to their ability to internalize in in vivo biological systems (i.e., cells). Although they can be harvested from various sources including stem cells, yet an appropriate isolation and characterization protocol to obtain "pure" exosomal population is needed. For potential clinical applications, understanding the functional ability of exosomes and their purity, that is, free from microvesicles, apoptotic bodies, and protein aggregates, is a pre-requisite. To achieve high purity and yield of exosomes from human Wharton's jelly-derived mesenchymal stem cells (hWJ-MSCs) in the size range of 30-200 nm, we have performed and compared three isolation procedures: ultracentrifugation (UC), sucrose cushion (SC), and commercially available reagent (CR). The isolated exosomes were characterized using nanoparticle tracking analysis (NTA), field emission scanning electron microscopy (FESEM), and atomic force microscopy (AFM). Furthermore, to understand the therapeutic potential of the hWJ-MSC-derived exosomes (hWJ-ME) to target pancreatic tumor cells, the internalization efficacy has been evaluated on the MiaPaCa-2 cell lines using confocal microscopy and flow cytometry. The NTA results showed sucrose cushion to be an optimal method for exosome isolation with high purity (86.8%), as compared to UC (40.5%; p = 0.050) and CR (38%; p = 0.050). Optical analysis by FESEM and AFM revealed that SC-isolated exosomes presented a spherical morphology, whereas UC- and CR-isolated exosomes exhibited an uneven morphology. Furthermore, the data from confocal images and flow cytometry showed that hWJ-ME were internalized by MiaPaCa-2, demonstrating the feasibility of exosomes as a "potential nanocarrier". Thus, our study suggests that a combination of NTA (yield), AFM (dimensions), and FESEM (morphology and topography) could provide sensitive biophysical characterization of hWJ-ME. In the future, enriched exosomes could be used as a delivery vehicle to transport target-specific drugs or gene-silencing constructs to tumors.
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Affiliation(s)
- Neha Chopra
- Department
of Research, Sir Ganga Ram Hospital, Old Rajinder Nagar, Delhi 110060, India
- Department
of Biotechnology, Jamia Hamdard, Hamdard Nagar, New Delhi 110062, India
| | - Braham Dutt Arya
- CSIR−National
Physical Laboratory, Dr. K. S. Krishnan Marg, New Delhi 110012, India
- Academy
of Scientific & Innovative Research (AcSIR), New Delhi 110025, India
| | - Namrata Jain
- Malvern
Panalytical Ltd., Enigma Business Park, Malvern WR14 1XZ, U.K.
| | - Poonam Yadav
- Department
of Research, Sir Ganga Ram Hospital, Old Rajinder Nagar, Delhi 110060, India
| | - Saima Wajid
- Department
of Biotechnology, Jamia Hamdard, Hamdard Nagar, New Delhi 110062, India
| | - Surinder P. Singh
- CSIR−National
Physical Laboratory, Dr. K. S. Krishnan Marg, New Delhi 110012, India
- Academy
of Scientific & Innovative Research (AcSIR), New Delhi 110025, India
- E-mail: (S.P.S.)
| | - Sangeeta Choudhury
- Department
of Research, Sir Ganga Ram Hospital, Old Rajinder Nagar, Delhi 110060, India
- E-mail: (S.C.)
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Selich A, Ha TC, Morgan M, Falk CS, von Kaisenberg C, Schambach A, Rothe M. Cytokine Selection of MSC Clones with Different Functionality. Stem Cell Reports 2019; 13:262-273. [PMID: 31303506 PMCID: PMC6700478 DOI: 10.1016/j.stemcr.2019.06.001] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/12/2018] [Revised: 06/07/2019] [Accepted: 06/10/2019] [Indexed: 12/26/2022] Open
Abstract
Mesenchymal stromal cells (MSCs) are used in many clinical applications. However, ex vivo expansion is required to reach clinically relevant cell numbers, which might lead to selection of clones with different characteristics. To follow clonal selection, we transduced MSC progenitors in umbilical cord pieces (UCPs) with vectors encoding fluorescent proteins and genetic barcodes. After marked MSC cultures grew out from UCPs, we investigated the influence of cytokines on MSC functionality. Specific cytokine conditions selected for clones from common progenitors. MSC secretome analyses revealed differences dependent on the culture conditions used. Clones expanded in human serum containing culture medium secreted a plethora of growth factors. When expanded in the same medium containing TGF-β, MSCs secreted negligible amounts of cytokines but at the same time led to an increased human chimerism after hematopoietic stem cell transplantation into immunodeficient mice. Our results suggest a major influence of cytokine additives on MSC functionality.
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Affiliation(s)
- Anton Selich
- Institute of Experimental Hematology, Hannover Medical School, Building J11, Level 01, Room 6540, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany
| | - Teng-Cheong Ha
- Institute of Experimental Hematology, Hannover Medical School, Building J11, Level 01, Room 6540, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany; Integrated Research and Treatment Center Transplantation (IFB-Tx), 30625 Hannover, Germany
| | - Michael Morgan
- Institute of Experimental Hematology, Hannover Medical School, Building J11, Level 01, Room 6540, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany; Integrated Research and Treatment Center Transplantation (IFB-Tx), 30625 Hannover, Germany; REBIRTH Cluster of Excellence, Hannover Medical School, 30625 Hannover, Germany
| | - Christine S Falk
- Integrated Research and Treatment Center Transplantation (IFB-Tx), 30625 Hannover, Germany; REBIRTH Cluster of Excellence, Hannover Medical School, 30625 Hannover, Germany; Institute of Transplant Immunology, Hannover Medical School, 30625 Hannover, Germany
| | | | - Axel Schambach
- Institute of Experimental Hematology, Hannover Medical School, Building J11, Level 01, Room 6540, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany; Integrated Research and Treatment Center Transplantation (IFB-Tx), 30625 Hannover, Germany; REBIRTH Cluster of Excellence, Hannover Medical School, 30625 Hannover, Germany; Division of Hematology/Oncology, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA
| | - Michael Rothe
- Institute of Experimental Hematology, Hannover Medical School, Building J11, Level 01, Room 6540, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany; REBIRTH Cluster of Excellence, Hannover Medical School, 30625 Hannover, Germany.
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30
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Pinto D, Bandeiras C, Fuzeta M, Rodrigues CAV, Jung S, Hashimura Y, Tseng R, Milligan W, Lee B, Ferreira FC, Silva C, Cabral JMS. Scalable Manufacturing of Human Mesenchymal Stromal Cells in the Vertical‐Wheel Bioreactor System: An Experimental and Economic Approach. Biotechnol J 2019; 14:e1800716. [DOI: 10.1002/biot.201800716] [Citation(s) in RCA: 21] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/06/2018] [Revised: 03/12/2019] [Indexed: 12/19/2022]
Affiliation(s)
- Diogo Pinto
- Department of Bioengineering and iBB, Institute for Bioengineering and Biosciences, Instituto Superior TécnicoUniversidade de LisboaAvenida Rovisco Pais Lisboa 1049‐001 Portugal
| | - Cátia Bandeiras
- Department of Bioengineering and iBB, Institute for Bioengineering and Biosciences, Instituto Superior TécnicoUniversidade de LisboaAvenida Rovisco Pais Lisboa 1049‐001 Portugal
- The Discoveries Center for Regenerative and Precision Medicine, Lisbon Campus, Instituto Superior TécnicoUniversidade de LisboaAvenida Rovisco Pais Lisboa 1049‐001 Portugal
- Division of Clinical Informatics, Department of MedicineBeth Israel Deaconess Medical Center1330 Beacon Street Brookline MA 02446 USA
| | - Miguel Fuzeta
- Department of Bioengineering and iBB, Institute for Bioengineering and Biosciences, Instituto Superior TécnicoUniversidade de LisboaAvenida Rovisco Pais Lisboa 1049‐001 Portugal
| | - Carlos A. V. Rodrigues
- Department of Bioengineering and iBB, Institute for Bioengineering and Biosciences, Instituto Superior TécnicoUniversidade de LisboaAvenida Rovisco Pais Lisboa 1049‐001 Portugal
- The Discoveries Center for Regenerative and Precision Medicine, Lisbon Campus, Instituto Superior TécnicoUniversidade de LisboaAvenida Rovisco Pais Lisboa 1049‐001 Portugal
| | - Sunghoon Jung
- PBS Biotech Inc1183 Calle Suerte Camarillo CA 93012 USA
| | - Yas Hashimura
- PBS Biotech Inc1183 Calle Suerte Camarillo CA 93012 USA
| | - Rong‐Jeng Tseng
- AventaCell Biomedical Corp., Global Center for Medical Innovation (GCMI)575 14th St NW Atlanta GA 30318 USA
| | - William Milligan
- AventaCell Biomedical Corp., Global Center for Medical Innovation (GCMI)575 14th St NW Atlanta GA 30318 USA
| | - Brian Lee
- PBS Biotech Inc1183 Calle Suerte Camarillo CA 93012 USA
| | - Frederico Castelo Ferreira
- Department of Bioengineering and iBB, Institute for Bioengineering and Biosciences, Instituto Superior TécnicoUniversidade de LisboaAvenida Rovisco Pais Lisboa 1049‐001 Portugal
- The Discoveries Center for Regenerative and Precision Medicine, Lisbon Campus, Instituto Superior TécnicoUniversidade de LisboaAvenida Rovisco Pais Lisboa 1049‐001 Portugal
| | - Cláudia Silva
- Department of Bioengineering and iBB, Institute for Bioengineering and Biosciences, Instituto Superior TécnicoUniversidade de LisboaAvenida Rovisco Pais Lisboa 1049‐001 Portugal
- The Discoveries Center for Regenerative and Precision Medicine, Lisbon Campus, Instituto Superior TécnicoUniversidade de LisboaAvenida Rovisco Pais Lisboa 1049‐001 Portugal
| | - Joaquim M. S. Cabral
- Department of Bioengineering and iBB, Institute for Bioengineering and Biosciences, Instituto Superior TécnicoUniversidade de LisboaAvenida Rovisco Pais Lisboa 1049‐001 Portugal
- The Discoveries Center for Regenerative and Precision Medicine, Lisbon Campus, Instituto Superior TécnicoUniversidade de LisboaAvenida Rovisco Pais Lisboa 1049‐001 Portugal
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Jara CP, do Prado TP, Dias Bóbbo VC, Ramalho ADFS, Lima MHM, Velloso LA, Araujo EP. Topical Topiramate Improves Wound Healing in an Animal Model of Hyperglycemia. Biol Res Nurs 2019; 21:420-430. [PMID: 31043061 DOI: 10.1177/1099800419845058] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/31/2022]
Abstract
Wound healing is severely affected in hyperglycemia and other metabolic conditions. Finding new therapeutic approaches that accelerate wound healing and improve the quality of the scar may reduce the morbidity commonly associated with skin lesions in diabetes. This study evaluated the effect of topical topiramate (TPM) on wound healing in C57 mice. Streptozotocin-induced hyperglycemic mice were subjected to a wound on the back and randomly allocated for treatment with either vehicle or topical TPM cream (2%) once a day for 14 days. Polymerase chain reaction, Western blotting, and microscopy were performed for the analysis. TPM improved wound healing (complete resolution at Day 10, 98% ± 5 for TPM vs. 81% ± 28 for vehicle), increased organization and deposition of collagen Type I, and enhanced the quality of the scars as determined by microscopy. In addition, TPM modulated the expression of cytokines and proteins of the insulin-signaling pathway: In early wound-healing stages, expression of interleukin-10, an anti-inflammatory marker, increased, whereas at the late phase, the pro-inflammatory markers tumor necrosis factor-α and monocyte chemoattractant protein-1 increased and there was increased expression of a vascular endothelial growth factor. Proteins of the insulin-signaling pathway were stimulated in the late wound-healing phase. Topical TPM improves the quality of wound healing in an animal model of hyperglycemia. The effect of TPM is accompanied by modulation of inflammatory and growth factors and proteins of the insulin-signaling pathway. Therefore, topical TPM presents as a potential therapeutic agent in skin wounds in patients with hyperglycemia.
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Affiliation(s)
- Carlos Poblete Jara
- 1 Nursing School, Laboratory of Cell Signaling, Obesity and Comorbidities Center (OCRC), University of Campinas, Campinas, São Paulo, Brazil
| | - Thais Paulino do Prado
- 1 Nursing School, Laboratory of Cell Signaling, Obesity and Comorbidities Center (OCRC), University of Campinas, Campinas, São Paulo, Brazil
| | - Vanessa Cristina Dias Bóbbo
- 1 Nursing School, Laboratory of Cell Signaling, Obesity and Comorbidities Center (OCRC), University of Campinas, Campinas, São Paulo, Brazil
| | - Albina de Fátima S Ramalho
- 1 Nursing School, Laboratory of Cell Signaling, Obesity and Comorbidities Center (OCRC), University of Campinas, Campinas, São Paulo, Brazil
| | - Maria H M Lima
- 1 Nursing School, Laboratory of Cell Signaling, Obesity and Comorbidities Center (OCRC), University of Campinas, Campinas, São Paulo, Brazil
| | - Licio A Velloso
- 1 Nursing School, Laboratory of Cell Signaling, Obesity and Comorbidities Center (OCRC), University of Campinas, Campinas, São Paulo, Brazil
| | - Eliana P Araujo
- 1 Nursing School, Laboratory of Cell Signaling, Obesity and Comorbidities Center (OCRC), University of Campinas, Campinas, São Paulo, Brazil
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Molecular and Functional Verification of Wharton's Jelly Mesenchymal Stem Cells (WJ-MSCs) Pluripotency. Int J Mol Sci 2019; 20:ijms20081807. [PMID: 31013696 PMCID: PMC6515095 DOI: 10.3390/ijms20081807] [Citation(s) in RCA: 33] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/14/2019] [Revised: 04/03/2019] [Accepted: 04/05/2019] [Indexed: 12/12/2022] Open
Abstract
The properties of mesenchymal stem cells (MSCs), especially their self-renewal and ability to differentiate into different cell lines, are widely discussed. Considering the fact that MSCs isolated from perinatal tissues reveal higher differentiation capacity than most adult MSCs, we examined mesenchymal stem cells isolated from Wharton's jelly of umbilical cord (WJ-MSCs) in terms of pluripotency markers expression. Our studies showed that WJ-MSCs express some pluripotency markers-such as NANOG, OCT-4, and SSEA-4-but in comparison to iPS cells expression level is significantly lower. The level of expression can be raised under hypoxic conditions. Despite their high proliferation potential and ability to differentiate into different cells type, WJ-MSCs do not form tumors in vivo, the major caveat of iPS cells. Owing to their biological properties, high plasticity, proliferation capacity, and ease of isolation and culture, WJ-MSCs are turning out to be a promising tool of modern regenerative medicine.
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Mahmoudian-Sani MR, Rafeei F, Amini R, Saidijam M. The effect of mesenchymal stem cells combined with platelet-rich plasma on skin wound healing. J Cosmet Dermatol 2018; 17:650-659. [PMID: 29504236 DOI: 10.1111/jocd.12512] [Citation(s) in RCA: 28] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 01/15/2018] [Indexed: 12/21/2022]
Abstract
INTRODUCTION Mesenchymal stem cells (MSCs) are multipotent stem cells that have the potential of proliferation, high self-renewal, and the potential of multilineage differentiation. The differentiation potential of the MSCs in vivo and in vitro has caused these cells to be regarded as potentially appropriate tools for wound healing. After the burn, trauma or removal of the tumor of wide wounds is developed. Although standard treatment for skin wounds is primary healing or skin grafting, they are not always practical mainly because of limited autologous skin grafting. EVIDENCE ACQUISITIONS Directory of Open Access Journals (DOAJ), Google Scholar, PubMed (NLM), LISTA (EBSCO), and Web of Science have been searched. EVIDENCE SYNTHESIS For clinical use of the MSCs in wound healing, two key issues should be taken into account: First, engineering biocompatible scaffolds clinical use of which leads to the least amount of side effects without any immunologic response and secondly, use of stem cells secretions with the least amount of clinical complications despite their high capability of healing damage. CONCLUSION In light of the MSCs' high capability of proliferation and multilineage differentiation as well as their significant role in modulating immunity, these cells can be used in combination with tissue engineering techniques. Moreover, the MSCs' secretions can be used in cell therapy to heal many types of wounds. The combination of MSCs and PRP aids wound healing which could potentially be used to promote wound healing.
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Affiliation(s)
| | - Fatemeh Rafeei
- Student Research Committee, Shahrekord University of Medical Sciences, Shahrekord, Iran
| | - Razieh Amini
- Department of Genetics and Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran
| | - Massoud Saidijam
- Department of Genetics and Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran
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Effects of topical topiramate in wound healing in mice. Arch Dermatol Res 2018; 310:363-373. [PMID: 29476247 DOI: 10.1007/s00403-018-1822-z] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/11/2017] [Revised: 01/02/2018] [Accepted: 02/15/2018] [Indexed: 02/07/2023]
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Yang L, Ma J, Gan S, Chu S, Maldonado M, Zhou J, Ma L, Tang S. Platelet poor plasma gel combined with amnion improves the therapeutic effects of human umbilical cord‑derived mesenchymal stem cells on wound healing in rats. Mol Med Rep 2017; 16:3494-3502. [PMID: 28714022 DOI: 10.3892/mmr.2017.6961] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/08/2016] [Accepted: 05/08/2017] [Indexed: 02/05/2023] Open
Abstract
The aim of the present study was to investigate the efficacy of human umbilical cord‑derived mesenchymal stem cell (HUMSCs) embedded in platelet poor plasma (PPP) gel combined with amnion (PPPA) in improving wound healing on Sprague‑Dawley (SD) rats. HUMSCs were cultured and labeled with chloromethylbenzamido‑1,1'‑dioctadecyl‑3,3,3'3'‑tetramethylindocarbocyanine perchlorate (CM‑DiI) on their third passage. The expression levels of growth factors of HUMSCs in PPPA were assessed by ELISA. Full‑thickness excisional skin wounds were induced in 36 male SD rats, which were treated with PPPA grafted with HUMSCs (PPPAC), PPPA, or HUMSC or PBS injection. The degree of healing and the distribution of labeled HUMSCs in the wound were evaluated by hematoxylin and eosin (H&E) staining and immunofluorescence. On day 14 post‑surgery, wound healing in PPPAC‑treated rats was significantly higher than the PPPA group, compared with rats treated with HUMSCs alone and control rats (P<0.05 and P<0.01, respectively). H&E staining showed that morphology and thickness of the epidermis in the PPPAC group was similar to that of healthy skin. ELISA revealed that levels of growth factors of HUMSCs in PPPAC were higher than in monolayer cells. In conclusion, PPPA can modify growth factor expression levels of HUMSCs and improve the efficiency of HUMSCs in the healing of full thickness wounds in rats.
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Affiliation(s)
- Lujun Yang
- Department of Burns and Plastic Surgery, The Second Affiliated Hospital, Shantou University Medical College, Shantou, Guangdong 515041, P.R. China
| | - Jie Ma
- Department of Burns and Plastic Surgery, The Second Affiliated Hospital, Shantou University Medical College, Shantou, Guangdong 515041, P.R. China
| | - Saiyang Gan
- Department of Burns and Plastic Surgery, The Second Affiliated Hospital, Shantou University Medical College, Shantou, Guangdong 515041, P.R. China
| | - Shuli Chu
- Department of Burns and Plastic Surgery, The Second Affiliated Hospital, Shantou University Medical College, Shantou, Guangdong 515041, P.R. China
| | - Martin Maldonado
- Research Center for Translational Medicine, Shantou University Medical College, Shantou, Guangdong 515041, P.R. China
| | - Jianda Zhou
- Department of Plastic Surgery, The Third Xiangya Hospital, Central South University, Changsha, Hunan 410013, P.R. China
| | - Lian Ma
- Research Center for Translational Medicine, Shantou University Medical College, Shantou, Guangdong 515041, P.R. China
| | - Shijie Tang
- Department of Burns and Plastic Surgery, The Second Affiliated Hospital, Shantou University Medical College, Shantou, Guangdong 515041, P.R. China
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Zorzopulos J, Opal SM, Hernando-Insúa A, Rodriguez JM, Elías F, Fló J, López RA, Chasseing NA, Lux-Lantos VA, Coronel MF, Franco R, Montaner AD, Horn DL. Immunomodulatory oligonucleotide IMT504: Effects on mesenchymal stem cells as a first-in-class immunoprotective/immunoregenerative therapy. World J Stem Cells 2017; 9:45-67. [PMID: 28396715 PMCID: PMC5368622 DOI: 10.4252/wjsc.v9.i3.45] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/12/2016] [Revised: 10/12/2016] [Accepted: 12/19/2016] [Indexed: 02/06/2023] Open
Abstract
The immune responses of humans and animals to insults (i.e., infections, traumas, tumoral transformation and radiation) are based on an intricate network of cells and chemical messengers. Abnormally high inflammation immediately after insult or abnormally prolonged pro-inflammatory stimuli bringing about chronic inflammation can lead to life-threatening or severely debilitating diseases. Mesenchymal stem cell (MSC) transplant has proved to be an effective therapy in preclinical studies which evaluated a vast diversity of inflammatory conditions. MSCs lead to resolution of inflammation, preparation for regeneration and actual regeneration, and then ultimate return to normal baseline or homeostasis. However, in clinical trials of transplanted MSCs, the expectations of great medical benefit have not yet been fulfilled. As a practical alternative to MSC transplant, a synthetic drug with the capacity to boost endogenous MSC expansion and/or activation may also be effective. Regarding this, IMT504, the prototype of a major class of immunomodulatory oligonucleotides, induces in vivo expansion of MSCs, resulting in a marked improvement in preclinical models of neuropathic pain, osteoporosis, diabetes and sepsis. IMT504 is easily manufactured and has an excellent preclinical safety record. In the small number of patients studied thus far, IMT504 has been well-tolerated, even at very high dosage. Further clinical investigation is necessary to demonstrate the utility of IMT504 for resolution of inflammation and regeneration in a broad array of human diseases that would likely benefit from an immunoprotective/immunoregenerative therapy.
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Scaling-Up Techniques for the Nanofabrication of Cell Culture Substrates via Two-Photon Polymerization for Industrial-Scale Expansion of Stem Cells. MATERIALS 2017; 10:ma10010066. [PMID: 28772424 PMCID: PMC5344595 DOI: 10.3390/ma10010066] [Citation(s) in RCA: 27] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 10/17/2016] [Revised: 01/05/2017] [Accepted: 01/10/2017] [Indexed: 01/28/2023]
Abstract
Stem-cell-based therapies require a high number (106–109) of cells, therefore in vitro expansion is needed because of the initially low amount of stem cells obtainable from human tissues. Standard protocols for stem cell expansion are currently based on chemically-defined culture media and animal-derived feeder-cell layers, which expose cells to additives and to xenogeneic compounds, resulting in potential issues when used in clinics. The two-photon laser polymerization technique enables three-dimensional micro-structures to be fabricated, which we named synthetic nichoids. Here we review our activity on the technological improvements in manufacturing biomimetic synthetic nichoids and, in particular on the optimization of the laser-material interaction to increase the patterned area and the percentage of cell culture surface covered by such synthetic nichoids, from a low initial value of 10% up to 88% with an optimized micromachining time. These results establish two-photon laser polymerization as a promising tool to fabricate substrates for stem cell expansion, without any chemical supplement and in feeder-free conditions for potential therapeutic uses.
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38
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Najar M, Lagneaux L. Foreskin as a source of immunotherapeutic mesenchymal stromal cells. Immunotherapy 2017; 9:207-217. [PMID: 28128711 DOI: 10.2217/imt-2016-0093] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/24/2016] [Accepted: 01/10/2017] [Indexed: 12/20/2022] Open
Abstract
Mesenchymal stromal cells (MSCs) have well-characterized properties and thus represent an attractive cell population for use in several therapeutic applications. Due to the limitations and inconveniences associated with classical sources of MSCs, the identification and characterization of alternative sources are required for safe and efficient cell therapy. The skin tissue is currently referred to as a reservoir of cells with therapeutically relevant functions. Historically considered biological waste, foreskin (FSK) is increasingly used to provide immunotherapeutic MSCs for medicinal products. This review discusses for the first time the nature and profile of MSCs within the foreskin tissue and, in particular, their immunobiology. A better immunological characterization and understanding of foreskin-derived cells will be critical for improving MSC-based cellular strategies for immunotherapeutic applications.
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Affiliation(s)
- Mehdi Najar
- Laboratory of Clinical Cell Therapy, Institut Jules Bordet, Université Libre de Bruxelles (ULB), Campus Erasme, Bâtiment de Transfusion (Level +1), Route de Lennik n° 808, 1070 Brussels, Belgium
| | - Laurence Lagneaux
- Laboratory of Clinical Cell Therapy, Institut Jules Bordet, Université Libre de Bruxelles (ULB), Campus Erasme, Bâtiment de Transfusion (Level +1), Route de Lennik n° 808, 1070 Brussels, Belgium
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39
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Bai T, Liu F, Zou F, Zhao G, Jiang Y, Liu L, Shi J, Hao D, Zhang Q, Zheng T, Zhang Y, Liu M, Li S, Qi L, Liu JY. Epidermal Growth Factor Induces Proliferation of Hair Follicle-Derived Mesenchymal Stem Cells Through Epidermal Growth Factor Receptor-Mediated Activation of ERK and AKT Signaling Pathways Associated with Upregulation of Cyclin D1 and Downregulation of p16. Stem Cells Dev 2016; 26:113-122. [PMID: 27702388 DOI: 10.1089/scd.2016.0234] [Citation(s) in RCA: 43] [Impact Index Per Article: 4.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/23/2022] Open
Abstract
The maintenance of highly proliferative capacity and full differentiation potential is a necessary step in the initiation of stem cell-based regenerative medicine. Our recent study showed that epidermal growth factor (EGF) significantly enhanced hair follicle-derived mesenchymal stem cell (HF-MSC) proliferation while maintaining the multilineage differentiation potentials. However, the underlying mechanism remains unclear. Herein, we investigated the role of EGF in HF-MSC proliferation. HF-MSCs were isolated and cultured with or without EGF. Immunofluorescence staining, flow cytometry, cytochemistry, and western blotting were used to assess proliferation, cell signaling pathways related to the EGF receptor (EGFR), and cell cycle progression. HF-MSCs exhibited surface markers of mesenchymal stem cells and displayed trilineage differentiation potentials toward adipocytes, chondrocytes, and osteoblasts. EGF significantly increased HF-MSC proliferation as well as EGFR, ERK1/2, and AKT phosphorylation (p-EGFR, p-ERK1/2, and p-AKT) in a time- and dose-dependent manner, but not STAT3 phosphorylation. EGFR inhibitor (AG1478), PI3K-AKT inhibitor (LY294002), ERK inhibitor (U0126), and STAT3 inhibitor (STA-21) significantly blocked EGF-induced HF-MSC proliferation. Moreover, AG1478, LY294002, and U0126 significantly decreased p-EGFR, p-AKT, and p-ERK1/2 expression. EGF shifted HF-MSCs at the G1 phase to the S and G2 phase. Concomitantly, cyclinD1, phosphorylated Rb, and E2F1expression increased, while that of p16 decreased. In conclusion, EGF induces HF-MSC proliferation through the EGFR/ERK and AKT pathways, but not through STAT-3. The G1/S transition was stimulated by upregulation of cyclinD1 and inhibition of p16 expression.
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Affiliation(s)
- Tingting Bai
- 1 Department of Pathobiology, Key Laboratory of Ministry of Education, College of Basic Medicine, Jilin University , Changchun, China .,2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China
| | - Feilin Liu
- 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China .,3 Department of Ophthalmology, Second Hospital of Jilin University , Changchun, China
| | - Fei Zou
- 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China .,4 Department of Pediatrics, First Hospital of Jilin University , Changchun, China
| | - Guifang Zhao
- 5 Department of Pathology, Jilin Medical College , Jilin, China
| | - Yixu Jiang
- 1 Department of Pathobiology, Key Laboratory of Ministry of Education, College of Basic Medicine, Jilin University , Changchun, China .,2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China
| | - Li Liu
- 1 Department of Pathobiology, Key Laboratory of Ministry of Education, College of Basic Medicine, Jilin University , Changchun, China .,2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China
| | - Jiahong Shi
- 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China
| | - Deshun Hao
- 1 Department of Pathobiology, Key Laboratory of Ministry of Education, College of Basic Medicine, Jilin University , Changchun, China .,2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China
| | - Qi Zhang
- 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China
| | - Tong Zheng
- 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China
| | - Yingyao Zhang
- 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China
| | - Mingsheng Liu
- 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China
| | - Shilun Li
- 6 Department of Oncology, First People's Hospital of Lishu County , Lishu County, China
| | - Liangchen Qi
- 7 Department of Thoracic Surgery, China-Japan Union Hospital of Jilin University , Changchun, China
| | - Jin Yu Liu
- 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China
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de Soure AM, Fernandes-Platzgummer A, da Silva CL, Cabral JMS. Scalable microcarrier-based manufacturing of mesenchymal stem/stromal cells. J Biotechnol 2016; 236:88-109. [PMID: 27527397 DOI: 10.1016/j.jbiotec.2016.08.007] [Citation(s) in RCA: 57] [Impact Index Per Article: 6.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/30/2016] [Revised: 08/02/2016] [Accepted: 08/09/2016] [Indexed: 12/17/2022]
Abstract
Due to their unique features, mesenchymal stem/stromal cells (MSC) have been exploited in clinical settings as therapeutic candidates for the treatment of a variety of diseases. However, the success in obtaining clinically-relevant MSC numbers for cell-based therapies is dependent on efficient isolation and ex vivo expansion protocols, able to comply with good manufacturing practices (GMP). In this context, the 2-dimensional static culture systems typically used for the expansion of these cells present several limitations that may lead to reduced cell numbers and compromise cell functions. Furthermore, many studies in the literature report the expansion of MSC using fetal bovine serum (FBS)-supplemented medium, which has been critically rated by regulatory agencies. Alternative platforms for the scalable manufacturing of MSC have been developed, namely using microcarriers in bioreactors, with also a considerable number of studies now reporting the production of MSC using xenogeneic/serum-free medium formulations. In this review we provide a comprehensive overview on the scalable manufacturing of human mesenchymal stem/stromal cells, depicting the various steps involved in the process from cell isolation to ex vivo expansion, using different cell tissue sources and culture medium formulations and exploiting bioprocess engineering tools namely microcarrier technology and bioreactors.
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Affiliation(s)
- António M de Soure
- Department of Bioengineering and iBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, Lisboa, Portugal
| | - Ana Fernandes-Platzgummer
- Department of Bioengineering and iBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, Lisboa, Portugal
| | - Cláudia L da Silva
- Department of Bioengineering and iBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, Lisboa, Portugal
| | - Joaquim M S Cabral
- Department of Bioengineering and iBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, Lisboa, Portugal.
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de Soure AM, Fernandes-Platzgummer A, Moreira F, Lilaia C, Liu SH, Ku CP, Huang YF, Milligan W, Cabral JMS, da Silva CL. Integrated culture platform based on a human platelet lysate supplement for the isolation and scalable manufacturing of umbilical cord matrix-derived mesenchymal stem/stromal cells. J Tissue Eng Regen Med 2016; 11:1630-1640. [DOI: 10.1002/term.2200] [Citation(s) in RCA: 33] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/21/2015] [Revised: 02/29/2016] [Accepted: 03/14/2016] [Indexed: 12/16/2022]
Affiliation(s)
- António M. de Soure
- Department of Bioengineering and iBB-; Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa; Lisboa Portugal
| | - Ana Fernandes-Platzgummer
- Department of Bioengineering and iBB-; Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa; Lisboa Portugal
| | - Francisco Moreira
- Department of Bioengineering and iBB-; Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa; Lisboa Portugal
| | - Carla Lilaia
- Hospital São Francisco Xavier, Centro Hospitalar Lisboa Ocidental; Lisboa Portugal
| | - Shi-Hwei Liu
- R&D, AventaCell BioMedical Co., Ltd.; New Taipei City Taiwan
| | - Chen-Peng Ku
- R&D, AventaCell BioMedical Co., Ltd.; New Taipei City Taiwan
- R&D, AventaCell BioMedical Co., Ltd; Atlanta GA USA
| | - Yi-Feng Huang
- R&D, AventaCell BioMedical Co., Ltd.; New Taipei City Taiwan
- R&D, AventaCell BioMedical Co., Ltd; Atlanta GA USA
| | - William Milligan
- R&D, AventaCell BioMedical Co., Ltd.; New Taipei City Taiwan
- R&D, AventaCell BioMedical Co., Ltd; Atlanta GA USA
| | - Joaquim M. S. Cabral
- Department of Bioengineering and iBB-; Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa; Lisboa Portugal
| | - Cláudia L. da Silva
- Department of Bioengineering and iBB-; Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa; Lisboa Portugal
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Mizukami A, Fernandes-Platzgummer A, Carmelo JG, Swiech K, Covas DT, Cabral JMS, da Silva CL. Stirred tank bioreactor culture combined with serum-/xenogeneic-free culture medium enables an efficient expansion of umbilical cord-derived mesenchymal stem/stromal cells. Biotechnol J 2016; 11:1048-59. [PMID: 27168373 DOI: 10.1002/biot.201500532] [Citation(s) in RCA: 56] [Impact Index Per Article: 6.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/10/2015] [Revised: 03/03/2016] [Accepted: 05/10/2016] [Indexed: 01/15/2023]
Abstract
Mesenchymal stem/stromal cells (MSC) are being widely explored as promising candidates for cell-based therapies. Among the different human MSC origins exploited, umbilical cord represents an attractive and readily available source of MSC that involves a non-invasive collection procedure. In order to achieve relevant cell numbers of human MSC for clinical applications, it is crucial to develop scalable culture systems that allow bioprocess control and monitoring, combined with the use of serum/xenogeneic (xeno)-free culture media. In the present study, we firstly established a spinner flask culture system combining gelatin-based Cultispher(®) S microcarriers and xeno-free culture medium for the expansion of umbilical cord matrix (UCM)-derived MSC. This system enabled the production of 2.4 (±1.1) x10(5) cells/mL (n = 4) after 5 days of culture, corresponding to a 5.3 (±1.6)-fold increase in cell number. The established protocol was then implemented in a stirred-tank bioreactor (800 mL working volume) (n = 3) yielding 115 million cells after 4 days. Upon expansion under stirred conditions, cells retained their differentiation ability and immunomodulatory potential. The development of a scalable microcarrier-based stirred culture system, using xeno-free culture medium that suits the intrinsic features of UCM-derived MSC represents an important step towards a GMP compliant large-scale production platform for these promising cell therapy candidates.
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Affiliation(s)
- Amanda Mizukami
- Hemotherapy Center of Ribeirão Preto, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto-SP, Brazil
| | - Ana Fernandes-Platzgummer
- Department of Bioengineering and IBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal
| | - Joana G Carmelo
- Department of Bioengineering and IBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal
| | - Kamilla Swiech
- Hemotherapy Center of Ribeirão Preto, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto-SP, Brazil. .,Department of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto-SP, Brazil.
| | - Dimas T Covas
- Hemotherapy Center of Ribeirão Preto, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto-SP, Brazil
| | - Joaquim M S Cabral
- Department of Bioengineering and IBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal
| | - Cláudia L da Silva
- Department of Bioengineering and IBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal .
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Aljawish A, Muniglia L, Chevalot I. Growth of human mesenchymal stem cells (MSCs) on films of enzymatically modified chitosan. Biotechnol Prog 2016; 32:491-500. [DOI: 10.1002/btpr.2216] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/31/2015] [Revised: 11/19/2015] [Indexed: 12/27/2022]
Affiliation(s)
- Abdulhadi Aljawish
- Université De Lorraine, Laboratoire D'ingénierie Des Biomolécules (LIBio); TSA40602-F-54518 Vandœuvre-lès-Nancy France
| | - Lionel Muniglia
- Université De Lorraine, Laboratoire D'ingénierie Des Biomolécules (LIBio); TSA40602-F-54518 Vandœuvre-lès-Nancy France
| | - Isabelle Chevalot
- Université De Lorraine, Laboratoire Réactions Et Génie Des Procédés (LRGP-CNRS-UMR 7274); TSA40602-F-54518 Vandœuvre-lès-Nancy France
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