Basic Study
Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Jan 26, 2015; 7(1): 195-207
Published online Jan 26, 2015. doi: 10.4252/wjsc.v7.i1.195
Early gestation chorionic villi-derived stromal cells for fetal tissue engineering
Lee Lankford, Taryn Selby, James Becker, Volodymyr Ryzhuk, Connor Long, Diana Farmer, Aijun Wang
Lee Lankford, Taryn Selby, James Becker, Volodymyr Ryzhuk, Connor Long, Diana Farmer, Aijun Wang, Department of Surgery, University of California, Davis Health System, Sacramento, CA 95817, United States
Author contributions: Lankford L, Selby T, Becker J and Ryzhuk V performed the experiments; Long C provided crucial assistance in data analysis and figure preparation; Lankford L, Farmer D and Wang A designed this study; all authors contributed to manuscript writing and editing.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (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: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Aijun Wang, PhD, Assistant Professor, Department of Surgery, University of California, Davis Health System, 4625 2nd Ave., Research II, Room 3005, Sacramento, CA 95817, United States. aawang@ucdavis.edu
Telephone: +1-916-7030422 Fax: +1-916-7030430
Received: September 5, 2014
Peer-review started: September 6, 2014
First decision: October 14, 2014
Revised: November 4, 2014
Accepted: November 7, 2014
Article in press: December 16, 2014
Published online: January 26, 2015
Processing time: 131 Days and 2.7 Hours
Abstract

AIM: To investigate the potential for early gestation placenta-derived mesenchymal stromal cells (PMSCs) for fetal tissue engineering.

METHODS: PMSCs were isolated from early gestation chorionic villus tissue by explant culture. Chorionic villus sampling (CVS)-size tissue samples (mean = 35.93 mg) were used to test the feasibility of obtaining large cell numbers from CVS within a clinically relevant timeframe. We characterized PMSCs isolated from 6 donor placentas by flow cytometry immunophenotyping, multipotency assays, and through immunofluorescent staining. Protein secretion from PMSCs was examined using two cytokine array assays capable of probing for over 70 factors in total. Delivery vehicle compatibility of PMSCs was determined using three common scaffold systems: fibrin glue, collagen hydrogel, and biodegradable nanofibrous scaffolds made from a combination of polylactic acid (PLA) and poly(lactic-co-glycolic acid) (PLGA). Viral transduction of PMSCs was performed using a Luciferase-GFP-containing lentiviral vector and efficiency of transduction was tested by fluorescent microscopy and flow cytometry analysis.

RESULTS: We determined that an average of 2.09 × 106 (SD ± 8.59 × 105) PMSCs could be obtained from CVS-size tissue samples within 30 d (mean = 27 d, SD ± 2.28), indicating that therapeutic numbers of cells can be rapidly expanded from very limited masses of tissue. Immunophenotyping by flow cytometry demonstrated that PMSCs were positive for MSC markers CD105, CD90, CD73, CD44, and CD29, and were negative for hematopoietic and endothelial markers CD45, CD34, and CD31. PMSCs displayed trilineage differentiation capability, and were found to express developmental transcription factors Sox10 and Sox17 as well as neural-related structural proteins NFM, Nestin, and S100β. Cytokine arrays revealed a robust and extensive profile of PMSC-secreted cytokines and growth factors, and detected 34 factors with spot density values exceeding 103. Detected factors had widely diverse functions that include modulation of angiogenesis and immune response, cell chemotaxis, cell proliferation, blood vessel maturation and homeostasis, modulation of insulin-like growth factor activity, neuroprotection, extracellular matrix degradation and even blood coagulation. Importantly, PMSCs were also determined to be compatible with both biological and synthetic material-based delivery vehicles such as collagen and fibrin hydrogels, and biodegradable nanofiber scaffolds made from a combination of PLA and PLGA. Finally, we demonstrated that PMSCs can be efficiently transduced (> 95%) with a Luciferase-GFP-containing lentiviral vector for future in vivo cell tracking after transplantation.

CONCLUSION: Our findings indicate that PMSCs represent a unique source of cells that can be effectively utilized for in utero cell therapy and tissue engineering.

Keywords: Placenta; Mesenchymal stromal cells; Chorionic villus; Fetal surgery; Tissue engineering

Core tip: In this study we characterize mesenchymal stromal cells derived from early gestation human placenta chorionic villi (PMSCs) for the purpose of fetal tissue engineering. We examine cell expansion in early passages from chorionic villus sampling-size tissue samples, as well as PMSC surface marker expression, multipotency, intracellular protein expression, protein secretion, and compatibility with delivery vehicles and tracking methods often used for in vivo experimentation. We show that early gestation PMSCs are excellent candidates for future tissue engineering studies, particularly as it applies to in utero therapy for congenital anomalies.