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Primary Cultures of Spermatogonia and Testis Cells

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Spermatogonial Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2656))

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Abstract

Spermatogonial stem cells (SSCs) maintain adult spermatogenesis in mammals by undergoing self-renewal and differentiation into spermatozoa. In order to study the biology of SSCs as related to spermatogenesis, an in vitro, long-term expansion system of SSCs constitutes an ideal tool. In this chapter, we describe a robust culture system for mouse and rat SSCs in vitro. In the presence of GDNF, GFRα1, and bFGF, SSCs maintained on STO feeder layers with serum-free medium continuously proliferate for over 6 months. Complete spermatogenesis in infertile recipient mice can be attained following transplantation of the cultured mouse and rat SSCs. Using the in vitro SSC culture systems, elucidation of stem cell biology can be advanced that significantly advances our understanding of spermatogenesis and male fertility.

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Acknowledgments

We thank Dr. Ralph Brinster and Dr. Eoin Whelan for helpful advice. This work was supported by the National Key R&D Program of China (Grant No. 2018YFC1003302) and the National Natural Science Foundation of China (Grant No. 31872844 and 32070831).

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Authors and Affiliations

  1. State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China

    Fan Yang, Jiachen Sun & Xin Wu

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  1. Fan Yang
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  2. Jiachen Sun
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  3. Xin Wu
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Correspondence to Xin Wu .

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Editors and Affiliations

  1. School of Molecular Biosciences, Washington State University, Pullman, WA, USA

    Jon M. Oatley

  2. Department of Neuroscience, Developmental and Regenerative Biology, University of Texas at San Antonio, San Antonio, TX, USA

    Brian P. Hermann

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Yang, F., Sun, J., Wu, X. (2023). Primary Cultures of Spermatogonia and Testis Cells. In: M. Oatley, J., Hermann, B.P. (eds) Spermatogonial Stem Cells. Methods in Molecular Biology, vol 2656. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3139-3_7

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  • DOI: https://doi.org/10.1007/978-1-0716-3139-3_7

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  • Print ISBN: 978-1-0716-3138-6

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