Male germ-like cell differentiation potential of human umbilical cord Wharton’s jelly-derived mesenchymal stem cells in co-culture with human placenta cells in presence of BMP4 and retinoic acid

Document Type: Original Article

Authors

1 Department of Reproductive Biology, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran

2 Department of Anatomical Sciences, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran

3 IVF Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran

4 Institute for Molecular Medicine and Cell Therapy, Düsseldorf, Germany and GENEOCELL, Advanced Molecular & Cellular Technologies, Tehran, Iran

5 Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran

6 Pediatric Urology Research Center, Children Hospital Medical Center, Tehran University of Medical Sciences, Tehran, Iran

7 Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

Abstract

Objective(s):Mesenchymal stem cells (MSCs) derived from Wharton’s jelly (WJ-MSCs) are now much more appealing for cell-based infertility therapy. Hence, WJ-MSCs differentiation toward germ layer cells for cell therapy purposes is currently under intensive study.
Materials and Methods: MSCs were isolated from human Wharton’s jelly and treated with BMP4, retinoic acid (RA) or co-cultured on human amniotic epithelial (HAE) and chorionic plate (HCP) placenta feeder cells. profile of POU5F1, Fragilis, Plzf, DDX4, Piwil2, Stra8, Dazl, β1- and α6-integrins (ITΒ1, ITA6) genes expression as germ cell markers were analyzed using RT-PCR and real-time PCR. Immunocytochemistry of surface markers were conducted.
Results: After 3 weeks treatment with different reagents and co-culture system, morphology of WJ-MSCs  changed to shiny clusters and germ cell specific markers in mRNA were up-regulated in both placental feeder + RA and BMP4 + RA. Induction of hWJ-MSCs with BMP4 in presence of RA resulted in significant up-regulation (P≤0.05) of all germ cell specific genes (c-Kit; 2.84±0.59, DDX4; 1.69±0.39, Piwil2; 1.14±0.21, Dazl; 0.65±0.25, α6 integrin; 1.26±0.53, β1 integrins; 1.18±0.65) compared to control and placental feeder cells + RA. Our results indicated that HAE and HCP followed by RA treatment were involved in human germ cell development.
Conclusion: We demonstrated that under the right conditions, hWJ-MSCs have the ability to differentiate to germ cells and this provides an excellent pattern to study infertility cause and treatment.

Keywords


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