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

摘要

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 , b 1- and a 6-integrins (ITB1, 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, a 6 integrin; 1.26 ± 0.53, b 1 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.