Basic Study
Copyright ©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Mar 26, 2020; 12(3): 203-221
Published online Mar 26, 2020. doi: 10.4252/wjsc.v12.i3.203
Efficient differentiation of vascular smooth muscle cells from Wharton’s Jelly mesenchymal stromal cells using human platelet lysate: A potential cell source for small blood vessel engineering
Panagiotis Mallis, Aggeliki Papapanagiotou, Michalis Katsimpoulas, Alkiviadis Kostakis, Gerasimos Siasos, Eva Kassi, Catherine Stavropoulos-Giokas, Efstathios Michalopoulos
Panagiotis Mallis, Catherine Stavropoulos-Giokas, Efstathios Michalopoulos, Hellenic Cord Blood Bank, Biomedical Research Foundation Academy of Athens, Athens 11527, Greece
Aggeliki Papapanagiotou, Gerasimos Siasos, Eva Kassi, Department of Biological Chemistry, Medical School, National and Kapodistrian Univesity of Athens, Athens 15772, Greece
Michalis Katsimpoulas, Alkiviadis Kostakis, Center of Experimental Surgery, Biomedical Research Foundation Academy of Athens, Athens 11527, Greece
Gerasimos Siasos, First Department of Cardiology, “Hippokration” Hospital, University of Athens Medical School, Athens 15231, Greece
Eva Kassi, First Department of Internal Medicine, Laiko Hospital, Medical School, National and Kapodistrian University of Athens, Athens 11527, Greece
Author contributions: Mallis P designed and carried out the whole experimental procedure of this study; Papanagiotou A, Siassos G and Kassi E supervised the differentiation procedures of MSCs into VSMCs; Katsimpoulas M and Kostakis A supervised the decellularization procedures of human umbilical arteries; Michalopoulos E designed, supervised and corrected the whole manuscript; Stavropoulos-Giokas C supervised and approved the whole study.
Institutional review board statement: This study was reviewed and approved by the Bioethics Committee of Biomedical Research Foundation Academy of Athens (Reference No. 1440, 20/11/2018).
Conflict-of-interest statement: The authors declare no conflict of interest.
Data sharing statement: Informed consent regarding the experiments of the current study was received from all mother participants.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
Corresponding author: Panagiotis Mallis, MSc, PhD, Hellenic Cord Blood Bank, Biomedical Research Foundation Academy of Athens, 4 Soranou Ephessiou Street, Athens 11527, Greece.
Received: November 1, 2019
Peer-review started: November 1, 2019
First decision: December 12, 2019
Revised: January 17, 2020
Accepted: January 31, 2020
Article in press: January 31, 2020
Published online: March 26, 2020

The development of fully functional small diameter vascular grafts requires both a properly defined vessel conduit and tissue-specific cellular populations. Mesenchymal stromal cells (MSCs) derived from the Wharton’s Jelly (WJ) tissue can be used as a source for obtaining vascular smooth muscle cells (VSMCs), while the human umbilical arteries (hUAs) can serve as a scaffold for blood vessel engineering.


To develop VSMCs from WJ-MSCs utilizing umbilical cord blood platelet lysate.


WJ-MSCs were isolated and expanded until passage (P) 4. WJ-MSCs were properly defined according to the criteria of the International Society for Cell and Gene Therapy. Then, these cells were differentiated into VSMCs with the use of platelet lysate from umbilical cord blood in combination with ascorbic acid, followed by evaluation at the gene and protein levels. Specifically, gene expression profile analysis of VSMCs for ACTA2, MYH11, TGLN, MYOCD, SOX9, NANOG homeobox, OCT4 and GAPDH, was performed. In addition, immunofluorescence against ACTA2 and MYH11 in combination with DAPI staining was also performed in VSMCs. HUAs were decellularized and served as scaffolds for possible repopulation by VSMCs. Histological and biochemical analyses were performed in repopulated hUAs.


WJ-MSCs exhibited fibroblastic morphology, successfully differentiating into “osteocytes”, “adipocytes” and “chondrocytes”, and were characterized by positive expression (> 90%) of CD90, CD73 and CD105. In addition, WJ-MSCs were successfully differentiated into VSMCs with the proposed differentiation protocol. VSMCs successfully expressed ACTA2, MYH11, MYOCD, TGLN and SOX9. Immunofluorescence results indicated the expression of ACTA2 and MYH11 in VSMCs. In order to determine the functionality of VSMCs, hUAs were isolated and decellularized. Based on histological analysis, decellularized hUAs were free of any cellular or nuclear materials, while their extracellular matrix retained intact. Then, repopulation of decellularized hUAs with VSMCs was performed for 3 wk. Decellularized hUAs were repopulated efficiently by the VSMCs. Biochemical analysis revealed the increase of total hydroyproline and sGAG contents in repopulated hUAs with VSMCs. Specifically, total hydroxyproline and sGAG content after the 1st, 2nd and 3rd wk was 71 ± 10, 74 ± 9 and 86 ± 8 μg hydroxyproline/mg of dry tissue weight and 2 ± 1, 3 ± 1 and 3 ± 1 μg sGAG/mg of dry tissue weight, respectively. Statistically significant differences were observed between all study groups (P < 0.05).


VSMCs were successfully obtained from WJ-MSCs with the proposed differentiation protocol. Furthermore, hUAs were efficiently repopulated by VSMCs. Differentiated VSMCs from WJ-MSCs could provide an alternative source of cells for vascular tissue engineering.

Keywords: Vascular smooth muscle cells, Decellularized umbilical arteries, Mesenchymal stromal cells, MYOCD, Cardiovascular disease, Blood vessels

Core tip: In this study, mesenchymal stromal cells derived from the Wharton’s Jelly tissue were differentiated into vascular smooth muscle cells (VSMCs). For this purpose, unlike the current literature, cord blood platelet lysate was used as the key element for the differentiation of mesenchymal stromal cells into VSMCs. Furthermore, the functional evaluation of VSMCs was tested. To do this, human umbilical arteries were decellularized and repopulated with the generated VSMCs.