title: Generation of male germ cells from induced pluripotent stem cells (iPS cells): an in vitro and in vivo study.
abstract: Recent studies have reported that induced pluripotent stem (iPS) cells from mice and humans can differentiate into primordial germ cells. However, whether iPS cells are capable of producing male germ cells is not known. The objective of this study was to investigate the differentiation potential of mouse iPS cells into spermatogonial stem cells and late-stage male germ cells. We used an approach that combines in vitro differentiation and in vivo transplantation. Embryoid bodies (EBs) were obtained from iPS cells using leukaemia inhibitor factor (LIF)-free medium. Quantitative PCR revealed a decrease in Oct4 expression and an increase in Stra8 and Vasa mRNA in the EBs derived from iPS cells. iPS cell-derived EBs were induced by retinoic acid to differentiate into spermatogonial stem cells (SSCs), as evidenced by their expression of VASA, as well as CDH1 and GFRα1, which are markers of SSCs. Furthermore, these germ cells derived from iPS cells were transplanted into recipient testes of mice that had been pre-treated with busulfan. Notably, iPS cell-derived SSCs were able to differentiate into male germ cells ranging from spermatogonia to round spermatids, as shown by VASA and SCP3 expression. This study demonstrates that iPS cells have the potential to differentiate into late-stage male germ cells. The derivation of male germ cells from iPS cells has potential applications in the treatment of male infertility and provides a model for uncovering the molecular mechanisms underlying male germ cell development.
authors.affiliation: Renji Hospital, Sperm Development and Genetics Laboratory, Shanghai Human Sperm Bank, Shanghai Institute of Andrology, Department of Urology, Shanghai Jiao Tong University School of Medicine, Shanghai 200001, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhu
authors.forename: Hong-Liang
authors.initials: HL
authors.lastname: Hu
authors.forename: Peng
authors.initials: P
authors.lastname: Li
authors.forename: Shi
authors.initials: S
authors.lastname: Yang
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Hui
authors.initials: H
authors.lastname: Ding
authors.forename: Ru-Hui
authors.initials: RH
authors.lastname: Tian
authors.forename: Ye
authors.initials: Y
authors.lastname: Ning
authors.forename: Ling-Ling
authors.initials: LL
authors.lastname: Zhang
authors.forename: Xi-Zhi
authors.initials: XZ
authors.lastname: Guo
authors.forename: Zhan-Ping
authors.initials: ZP
authors.lastname: Shi
authors.forename: Zheng
authors.initials: Z
authors.lastname: Li
authors.forename: Zuping
authors.initials: Z
authors.lastname: He
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D002066:Busulfan / Q000494:pharmacology; D064200:Cdh1 Proteins; D018797:Cell Cycle Proteins / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D053487:DEAD-box RNA Helicases / Q000378:metabolism; D004268:DNA-Binding Proteins; D058732:Embryoid Bodies / Q000378:metabolism; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009687:Nuclear Proteins / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012671:Seminiferous Tubules / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D013093:Spermatogonia / Q000166:cytology* / Q000637:transplantation; D013094:Spermatozoa / Q000166:cytology*; D013737:Testis / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D064200:Cdh1 Proteins; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; C577585:Fzr1 protein, mouse; C494988:Gfra1 protein, mouse; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors; D009687:Nuclear Proteins; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D011506:Proteins; D012333:RNA, Messenger; C102019:Stra8 protein, mouse; C490062:Sycp3 protein, mouse; D014212:Tretinoin; C090864:Ddx4 protein, mouse; D053487:DEAD-box RNA Helicases; D002066:Busulfan