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Differentiation of spermatogonial stem cell-like cells from murine testicular tissue into haploid male germ cells in vitro

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Abstract

In vitro differentiation of spermatogonial stem cells (SSCs) promotes the understanding of the mechanism of spermatogenesis. The purpose of this study was to isolate spermatogonial stem cell-like cells from murine testicular tissue, which then were induced into haploid germ cells by retinoic acid (RA). The spermatogonial stem cell-like cells were purified and enriched by a two-step plating method based on different adherence velocities of SSCs and somatic cells. Cell colonies were present after culture in M1-medium for 3 days. Through alkaline phosphatase, RT-PCR and indirect immunofluorescence cell analysis, cell colonies were shown to be SSCs. Subsequently, cell colonies of SSCs were cultured in M2-medium containing RA for 2 days. Then the cell colonies of SSCs were again cultured in M1-medium for 6–8 days, RT-PCR and indirect immunofluorescence cell analysis were chosen to detect haploid male germ cells. It could be demonstrated that 10−7 mol l−1 of RA effectively induced the SSCs into haploid male germ cells in vitro.

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Acknowledgments

This research was supported by the Youth Extra Fund of Northwest A&F University (Z111020905), the Basic Scientific Research Expense of Sci-Tech Innovation Major Project of Northwest A&F University (QN2011061) and the Special Research Subsidy Project of Northwest A&F University (07ZR002).

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  1. College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, People’s Republic of China

    Peng Wang, Li-Juan Suo, Hua Shang, Ying Li, Guang-Xuan Li, Qing-Wang Li & Jian-Hong Hu

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  1. Peng Wang
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  2. Li-Juan Suo
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  3. Hua Shang
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  4. Ying Li
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  5. Guang-Xuan Li
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  6. Qing-Wang Li
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  7. Jian-Hong Hu
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Correspondence to Jian-Hong Hu.

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Wang, P., Suo, LJ., Shang, H. et al. Differentiation of spermatogonial stem cell-like cells from murine testicular tissue into haploid male germ cells in vitro. Cytotechnology 66, 365–372 (2014). https://doi.org/10.1007/s10616-013-9584-0

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  • DOI: https://doi.org/10.1007/s10616-013-9584-0

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