title: Differentiation of induced pluripotent stem cells into male germ cells in vitro through embryoid body formation and retinoic acid or testosterone induction.
abstract: Generation of germ cells from pluripotent stem cells in vitro could 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 cells in 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 of Stra8, Odf2, Act, and Prm1 was upregulated in iPS cells by retinoic acid or testosterone induction, whereas Oct-4 transcription 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 cells in vitro through embryoid body formation and retinoic acid or testosterone induction.
authors.affiliation: Shanghai Human Sperm Bank, Department of Urology, Shanghai Institute of Andrology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200001, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Li
authors.forename: Hongliang
authors.initials: H
authors.lastname: Hu
authors.forename: Shi
authors.initials: S
authors.lastname: Yang
authors.forename: Ruhui
authors.initials: R
authors.lastname: Tian
authors.forename: Zhenzhen
authors.initials: Z
authors.lastname: Zhang
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Meng
authors.initials: M
authors.lastname: Ma
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhu
authors.forename: Xizhi
authors.initials: X
authors.lastname: Guo
authors.forename: Yiran
authors.initials: Y
authors.lastname: Huang
authors.forename: Zuping
authors.initials: Z
authors.lastname: He
authors.forename: Zheng
authors.initials: Z
authors.lastname: Li
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D058732:Embryoid Bodies / Q000166:cytology*; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D005854:Germ Cells / Q000166:cytology*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007246:Infertility / Q000628:therapy; D008297:Male; D051379:Mice; D008856:Microscopy, Fluorescence; D060888:Real-Time Polymerase Chain Reaction; D013739:Testosterone / Q000494:pharmacology*; D014212:Tretinoin / Q000494:pharmacology*
chemical_list: D013739:Testosterone; D014212:Tretinoin