Differentiation of Induced Pluripotent Stem Cells into Male Germ CellsIn Vitrothrough Embryoid Body Formation and Retinoic Acid or Testosterone Induction

Abstract

Generation of germ cells from pluripotent stem cellsin vitrocould have great application for treating infertility and provides an excellent model for uncovering molecular mechanisms controlling gametogenesis. In this study, we explored the differentiation potential of mouse induced pluripotent stem (iPS) cells towards male germ cells. Embryoid body formation and retinoic acid/testosterone induction were applied to promote differentiation of mouse iPS cells into male germ cellsin vitro. Quantitative RT-PCR and immunoflourescence were performed to characterize the iPS cell differentiation process, and notably there were different temporal expression profiles of male germ cell-associated genes. The expression of proteins, including MVH, CDH1, and SCP3, was remarkably increased. mRNA expression ofStra8,Odf2,Act, andPrm1was upregulated in iPS cells by retinoic acid or testosterone induction, whereasOct-4transcription was reduced in these cells compared to the controls. Hormones were also measured in the EB medium. DNA content analysis by flow cytometry revealed that iPS cells could differentiate into haploid cells through retinoic acid or testosterone treatment. Collectively, our results suggest that mouse iPS cells possess the potency to differentiate into male germ cellsin vitrothrough embryoid body formation and retinoic acid or testosterone induction.