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Reconstitution of male germline cell specification from mouse embryonic stem cells using defined factors in vitro

Cell Death & Differentiation (2019) | Download Citation

Abstract

In vitro induction of functional haploid cells from embryonic stem cells (ESCs) has been reported by several groups. However, these reports either involve complex induction process with undefined induction factors or show low-induction efficiency. Here, we report complete meiosis in vitro from ESCs with defined induction factors. ESCs were first induced into primordial germ cell-like cells, which were further induced into male germline cells, including spermatogonial stem cell-like cells (SSCLCs) and spermatid-like cells. Importantly, the obtained SSCLCs were functional as infertile male mice sired healthy offspring via SSCLC transplantation. Further, we found that eukaryotic translation initiation factor 2 subunit 3 and structural gene Y-linked (Eif2s3y) was essential for spermatogenesis. Eif2s3y-overexpressing ESCs showed enhanced spermatogenesis in vitro, as demonstrated by higher expression levels of SSC-specific markers during SSCLC induction process, improved reproductive ability recovery of infertile male mice, and increased efficiency of haploid cell induction. Our work provides a convenient and efficient approach to obtain functional male germline cells.

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Acknowledgments

We appreciate Dr. Christian Dani of University of Nice Sophia Antipolis for excellent comments and revisions. This work was supported by grants from the China National Basic Research Program (JFYS 2016YFA0100203), Doctoral Scientific Research Foundation of 2017 (Z109021715), the Program of National Natural Science Foundation of China (31572399 and 31272518) and China Postdoctoral Science Foundation (K3080217064).

Author contributions

N.L., W.T.M., Q.Y.S., M.F.Z., and W.Q.L. performed the experiments. N.L., W.T.M., Z.Y.D., C.Y.W., and B.W.N. analyzed the data. J.L.H. designed the project. N.L. and W.T.M. wrote the manuscript, and J.L.H. revised the manuscript.

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Affiliations

  1. College of Veterinary Medicine, Shaanxi Center of Stem Cells Engineering & Technology, Northwest A & F University, Yangling, Shaanxi, China

    • Na Li
    • , Wentao Ma
    • , Qiaoyan Shen
    • , Mengfei Zhang
    • , Zhaoyu Du
    • , Chongyang Wu
    • , Bowen Niu
    • , Wenqing Liu
    •  & Jinlian Hua

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The authors declare that they have no conflict of interest.

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Correspondence to Jinlian Hua.

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DOI

https://doi.org/10.1038/s41418-019-0280-2