In Vitro Modeling of Human Germ Cell Development Using Pluripotent Stem Cells

Yuncheng Zhao; Shicheng Ye; Dongli Liang; Pengxiang Wang; Jing Fu; Qing Ma; Ruijiao Kong; Linghong Shi; Xueping Gong; Wei Chen; Wubin Ding; Wenjing Yang; Zijue Zhu; Huixing Chen; Xiaoxi Sun; Jun Zhu; Zheng Li; Yuan Wang

doi:10.1016/j.stemcr.2018.01.001

In Vitro Modeling of Human Germ Cell Development Using Pluripotent Stem Cells

Stem Cell Reports. 2018 Feb 13;10(2):509-523. doi: 10.1016/j.stemcr.2018.01.001. Epub 2018 Feb 1.

Authors

Yuncheng Zhao¹, Shicheng Ye¹, Dongli Liang¹, Pengxiang Wang¹, Jing Fu², Qing Ma¹, Ruijiao Kong¹, Linghong Shi¹, Xueping Gong¹, Wei Chen¹, Wubin Ding¹, Wenjing Yang³, Zijue Zhu⁴, Huixing Chen⁴, Xiaoxi Sun², Jun Zhu³, Zheng Li⁴, Yuan Wang⁵

Affiliations

¹ Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, 500 Dongchuan Road, Shanghai 200241, China.
² Obstetrics and Gynecology Hospital of Fudan University, Shanghai 200011, China.
³ Systems Biology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
⁴ Department of Andrology, Urologic Medical Center, Shanghai Key Lab of Reproductive Medicine, Shanghai General Hospital, Shanghai Jiaotong University, Shanghai 200080, China.
⁵ Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, 500 Dongchuan Road, Shanghai 200241, China; Department of Animal Science, Michigan State University, Lansing, MI 48824, USA. Electronic address: ywang@bio.ecnu.edu.cn.

Abstract

Due to differences across species, the mechanisms of cell fate decisions determined in mice cannot be readily extrapolated to humans. In this study, we developed a feeder- and xeno-free culture protocol that efficiently induced human pluripotent stem cells (iPSCs) into PLZF+/GPR125+/CD90+ spermatogonium-like cells (SLCs). These SLCs were enriched with key genes in germ cell development such as MVH, DAZL, GFRα1, NANOS3, and DMRT1. In addition, a small fraction of SLCs went through meiosis in vitro to develop into haploid cells. We further demonstrated that this chemically defined induction protocol faithfully recapitulated the features of compromised germ cell development of PSCs with NANOS3 deficiency or iPSC lines established from patients with non-obstructive azoospermia. Taken together, we established a powerful experimental platform to investigate human germ cell development and pathology related to male infertility.

Keywords: infertility; non-obstructive azoospermia; pluripotent stem cells; spermatogonia.

Publication types

Research Support, N.I.H., Intramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Azoospermia / genetics*
Azoospermia / pathology
Cell Culture Techniques
Cell Differentiation / genetics
Feeder Cells / cytology
Gene Expression Regulation, Developmental / genetics
Germ Cells / cytology
Germ Cells / growth & development
Glial Cell Line-Derived Neurotrophic Factor Receptors / genetics
Humans
Induced Pluripotent Stem Cells / cytology*
Induced Pluripotent Stem Cells / pathology
Infertility, Male / genetics*
Infertility, Male / pathology
Male
Meiosis / genetics
Mice
Mouse Embryonic Stem Cells / cytology
RNA-Binding Proteins / genetics
Species Specificity
Spermatogonia / cytology
Spermatogonia / growth & development*
Spermatogonia / pathology
Transcription Factors / genetics

Substances

DAZL protein, human
DMRT1 protein
Gfra1 protein, mouse
Glial Cell Line-Derived Neurotrophic Factor Receptors
NANOS3 protein, human
RNA-Binding Proteins
Transcription Factors

Abstract

Publication types

MeSH terms

Substances

Grants and funding