Molecular Mechanism Of WJ-MSCs’ Differentiation Into Endometrial Cells

Objective Wharton’s jelly-derived mesenchymal stem cells(WJ-MSCs)are a novel and promising strategy for tissue engineering because of their ability to differentiate into many cell types.We characterized the differentiation of WJ-MSCs into endometrial stromal cell(ESC)-like cells.we further explored the molecular mechanisms underlying the differentiation of WJ-MSCs into ESC-like cells.Methods WJ-MSCs were treated in two ways respectively to differentiate into EEC-like and ESC-like cells: 1.Co-cultured with ESCs in control/differentiation medium(17β-estradiol,growth factors);2.Cultured in control/differentiation medium(8-Br-cAMP alone or 8-Br-cAMP plus 17-βestrogen and growth factors).Three signaling pathway inhibitors(SB203580,PD98059,H89)were used to investigate the mechanism of WJ-MSCs differentiation into ESC-like cells.Western blotting and flow cytometry were used to analyze expression of epithelial markers and stromal cell markers.Enzyme-linked immunosorbent assays were used to test the production of secretory proteins associated with the differentiation of ESC-like cells.Results 8-Br-cAMP at 0.5 mM upregulated vimentin and CD13 and downregulated CK and CD9,promoting the differentiation of WJ-MSCs into ESC-like cells.PRL and IGFBP1 were upregulated and the PKA signaling pathway was activated,whereas ERK1/2 and p38 MAPK were not affected.WJ-MSCs didn’t differentiated into EEC-like cells in the coculture system.Conclusions 8-Br-cAMP plus estrogen and growth factors can induce the differentiation of WJ-MSCs into ESC-like cells.During the differentiation of WJ-MSCs into ESC-like cells,PRL and IGFBP1 were upregulated by the treatment and the PKA signaling pathway was activated,whereas ERK1/2 and p38 MAPK were not affected.These findings suggest a promising approach to the treatment of endometrial damage and other endometrial diseases and suggest new applications for WJ-MSCs in clinical practice.