BMP۴ Cannot Be Substituted by Small-Molecule During Germ Cell Differentiation from Embryonic Stem Cells

Objective: Germ cell production from stem cells allows inves-tigation of the involved mechanisms in gametes development with the goal of producing healthy gametes for infertile cou-ples. In this regard, improvement of the protocols for in vitro reconstitution of germ cell development from stem cells is in the spotlight of research. Recently, SB۴ small molecule has been introduced as a potent agonist for BMP۴. These evidences suggest SB۴ as a potent candidate to substitute BMP۴ (as key inducer of germ cell) during in vitro germ cell differentiation for providing a cost-effective protocol.Materials and Methods: To examine the SB۴ as a substitute for BMP۴ during in vitro germ cell differentiation, after find-ing non-toxically SB۴ concentration for germ cell culture by PI staining (۲۰۰ µM/ml), germ cells were induced via Hayashi’s protocol in the presence of BMP۴ or SB۴ or both of them. Ag-gregates were collected for gene expression analysis after ۴ days of induction.Results: Our data showed that the level of Blimp۱ expression in SB۴-induced aggregates is significantly less, compared to BMP۴-induced aggregates (P<۰.۰۵). Prdm۱۴ expression in-creased significantly in both SB۴- and BMP۴- induced cells compared (P<۰.۰۱), but no significant differences were shown in SB۴- and BMP۴- induced groups. Also, at the simultaneous presence of BMP۴ and SB۴, there was significantly increasing levels of Prdm۱۴ but no significant difference in Blimp۱ ex-pression compared to BMP۴ alone.Conclusion: It seems that the SB۴ could not induce germ cell lineage. However Simultaneous presence of BMP۴ and SB۴ may improve the establishment of germ cell fate in vitro via increasing Prdm۱۴ expression.

کلیدواژه ها:

Infertility ، Primordial Germ Cells ، BMP۴ ، Small Molecule ، SB۴