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β-estradiol promotes the growth of primary human fetal spermatogonial stem cells via the induction of stem cell factor in Sertoli cells

  • Gamete Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Background

Mammalian spermatogenesis is responsible for male fertility and is supported by the self-renewal and differentiation of spermatogonial stem cells (SSCs). Sertoli cells provide a supportive microenvironment for SSCs, in part by the production of stem cell factor (SCF), which is a potent regulator of spermatogonia proliferation and survival.

Methods

We investigated the novel role of β-estradiol in modulating the proliferation and apoptosis of fetal SSCs via the regulation of SCF secretion in Sertoli cells isolated from human fetal testes. The proliferation of SSCs in the co-culture system was determined by colony formation and BrdU incorporation assays. TUNEL assay was used to measure SSC apoptosis in co-culture in response to treatment with control, β-estradiol, or the combination of β-estradiol and the estrogen receptor inhibitor ICI 182780.

Results

In the system with purified human fetal Sertoli cells (MIS+/c-Kit−/AP−), β-estradiol upregulated the production of SCF in a dose- and time-dependent manner. In the co-culture system of primary human fetal SSCs (c-Kit+/SSEA-4+/Oct-4+/AP+) and Sertoli cells (MIS+), β-estradiol markedly increased the proliferation of SSCs. Moreover, SSC apoptosis was significantly inhibited by β-estradiol and was completely reversed by the combination of β-estradiol and ICI 182780.

Conclusion

Here we report, for the first time, that β-estradiol can induce the increase of SCF expression in human fetal Sertoli cells and regulates the growth and survival of human fetal SSCs. These novel findings provide new perspectives on the current understanding of the role of estrogen in human spermatogenesis.

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Funding

This work was supported by the National Natural Science Foundation of China (No. 31472054), the National Key Research and Development Program of China (2016YFC1000600), the Natural Science Foundation of Hunan Province (No. 2018JJ3367), and the Scientific Research Foundation of Hunan Provincial Education Department (No. 18C0067).

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Authors and Affiliations

  1. Institute of Reproductive and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha, 410078, China

    Ke Tao, Yuan Sun, Yuanchi Chao, Liu Xing, Lizhi Leng, Dai Zhou, Wenbing Zhu & Liqing Fan

  2. Department of Medical Laboratory, School of Medicine, Hunan Normal University, Changsha, 410013, China

    Ke Tao

  3. Reproductive & Genetic Hospital of CITIC-Xiangya, Changsha, 410078, China

    Liu Xing, Lizhi Leng, Dai Zhou, Wenbing Zhu & Liqing Fan

Authors
  1. Ke Tao
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  2. Yuan Sun
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  3. Yuanchi Chao
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  4. Liu Xing
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  5. Lizhi Leng
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  6. Dai Zhou
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  7. Wenbing Zhu
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  8. Liqing Fan
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Contributions

LQF and KT designed the study. YCC, YS, LX, and DZ supervised sample collection. KT, YS, and WBZ performed the experiments and data collection. LZL revised data analysis and figures preparation. All authors have participated in the manuscript preparation.

Corresponding author

Correspondence to Liqing Fan.

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Ethics approval

This study was approved by the Ethics Committee of the Reproductive & Genetic Hospital of CITIC-Xiangya, Basic Medical Science School, Central South University, Changsha, China.

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Oral and written informed consent was obtained from each participant.

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The authors declare no competing interests.

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Supplementary information

Fig. S1.

Identification of fetal SSCs and Sertoli cells in 20-week-old human fetal testes. Immunofluorescence analysis was performed on a cryosection of a 20-week-old human fetal testis. (a) Immunostaining of Oct4 (green); (b) immunostaining of MIS (green). The nuclei are stained with DAPI (blue). Scale bar: 50 μm. (PNG 267 kb)

High resolution image (TIF 4825 kb)

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Tao, K., Sun, Y., Chao, Y. et al. β-estradiol promotes the growth of primary human fetal spermatogonial stem cells via the induction of stem cell factor in Sertoli cells. J Assist Reprod Genet 38, 2481–2490 (2021). https://doi.org/10.1007/s10815-021-02240-y

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