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Asian Journal of Andrology (2012) 14, 574-579; doi:10.1038/aja.2012.3; published online 16 April 2012
Generation of male germ cells from induced pluripotent stem cells (iPS cells): an in vitro and in vivo study
Yong Zhu1,*, Hong-Liang Hu1,*, Peng Li1, Shi Yang1, Wei Zhang1, Hui Ding1, Ru-Hui Tian1, Ye Ning1, Ling-Ling Zhang2, Xi-Zhi Guo2, Zhan-Ping Shi1, Zheng Li1 and Zuping He3,4
1 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
2 Bio-X Institutes, Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Shanghai Jiao Tong University, Shanghai 200240, China
3 Clinic Stem Cell Research Center, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China
4 Department of Biochemistry and Molecular and Cellular Biology, Georgetown University Medical Center, Washington, DC 20057, USA
Correspondence: Professor Z Li, (doc.zheng.li@gmail.com); Professor Z He, (zupinghe@gmail.com)
* The authors contributed equally to this article.
Received 16 September 2011; Revised 22 December 2011; Accepted 15 January 2012
Advance online publication 16 April 2012.
| 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.
Keywords: differentiation; induced pluripotent stem cells; male germ cells; retinoic acid; transplantation |
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