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Leptin promotes proliferation of neonatal mouse stem/progenitor spermatogonia

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

Abstract

Purpose

To keep and increase spermatogonial stem cell number (SSC) is the only available option for pediatric cancer survivors to maintain fertility. Leptin is secreted by the epididymal white adipose tissue and has receptors on stem/progenitor spermatogonia. The purpose of this study is to demonstrate dose- and time-dependent proliferative effect of leptin on stem/progenitor spermatogonia cultures from prepubertal mice testes.

Methods

CD90.2 (+) stem/progenitor spermatogonia were isolated from the C57BL/6 mouse testis on postnatal day 6 and placed in culture. The proliferative effect of leptin supplementation was assessed by colony formation (diameter and number), WST proliferation assays, and xCELLigence real-time cell analysis (RTCA) on days 3, 5, and 7 of culture. Expressions of p-ERK1/2, p-STAT3, total STAT3, and p-SHP2 levels were determined by western blot analysis.

Results

Leptin supplementation of 100 ng/ml increased the diameter (p = 0.001) and number (p = 0.01) of colonies in stem/progenitor spermatogonial cultures and caused higher proliferation by WST-1 (p = 0.009) compared with the control on day 7. The EC50 was calculated as 114 ng/ml for leptin by RTCA. Proliferative dose of leptin induced increased expression of p-ERK1/2 (p = 0.009) and p-STAT3 (p = 0.023) on stem/progenitor spermatogonia when compared with the untreated group.

Conclusion

The results indicated that leptin supplementation exhibited a dose- and time-dependent proliferative effect on stem/progenitor spermatogonia that was associated with increased expression of ERK1/2 and STAT3 pathways while maintaining their undifferentiated state. This output presents a new agent that may help to expand the stem/progenitor spermatogonia pool from the neonatal testis in order to autotransplant after cancer treatment.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

The Research Coordination Unit of Hacettepe University funded this work (THD-2017-13430). Professor Kyle E Orwig was also supported by the Eunice Kennedy Shriver National Institute of Child Health and Human Development grant HD092084.

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Authors and Affiliations

  1. Department of Histology and Embryology, Hacettepe University Faculty of Medicine, Ankara, Turkey

    Nilgün Yersal & Petek Korkusuz

  2. Department of Medical Biology, Atılım University Faculty of Medicine, Ankara, Turkey

    Sevil Köse

  3. Department of Basic Oncology, Hacettepe University Cancer Institute, Ankara, Turkey

    Utku Horzum

  4. Center for Assisted Reproduction, Department of Obstetrics and Gynecology, Ankara University Faculty of Medicine, Ankara, Turkey

    Sinan Özkavukcu

  5. Department of Obstetrics, Gynecology and Reproductive Sciences, Magee-Womens Research Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213, USA

    Kyle E. Orwig

Authors
  1. Nilgün Yersal
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  2. Sevil Köse
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  3. Utku Horzum
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  4. Sinan Özkavukcu
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  5. Kyle E. Orwig
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  6. Petek Korkusuz
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Contributions

Nilgun Yersal generated the hypothesis, established the rationale and designed the study together with her mentor Petek Korkusuz, and wrote the manuscript. Nilgun Yersal performed the experimental study, cultured the stem/progenitor spermatogonia, and performed histological examination of the testes. Utku Horzum analyzed and interpreted the flow cytometry and western blotting data. Sevil Kose supervised the MACS isolation and interpreted the xCELLigence data. Sinan Ozkavukcu contributed to the analysis of the cell data. Dr Kyle E Orwig, as the co-advisor of Nilgun Yersal, supervised the Ob receptor immune labeling; Petek Korkusuz and Kyle E Orwig edited the manuscript. All authors read and approved the final manuscript

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Correspondence to Petek Korkusuz.

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The Hacettepe University Animal Experimentations Local Ethics Board (#2016/59/1) approved the use of animal material.

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Yersal, N., Köse, S., Horzum, U. et al. Leptin promotes proliferation of neonatal mouse stem/progenitor spermatogonia. J Assist Reprod Genet 37, 2825–2838 (2020). https://doi.org/10.1007/s10815-020-01929-w

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