Leptin promotes proliferation of neonatal mouse stem/progenitor spermatogonia

Nilgun Yersal, Sevil Kose, Utku Horzum, Sinan Ozkavukcu, Kyle E. Orwig, Petek Korkusuz

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)


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.
Original languageEnglish
Pages (from-to)2825-2838
Number of pages14
JournalJournal of Assisted Reproduction and Genetics
Issue number11
Early online date25 Aug 2020
Publication statusPublished - Nov 2020


  • Stem/progenitor spermatogonia
  • Leptin
  • Proliferation
  • p-STAT3
  • p-ERK1/2


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