Abstract
In the present study, we used a serum-free culture media to propagate goat putative spermatogonial stem cells (SSCs) and evaluated the effect of crucial growth factors on relative expression of some SSC markers and self-renewal related genes. The enriched SSCs were cultured on a homologous Sertoli cell feeder layer in KO-DMEM supplemented with 10% KOSR. Putative SSC colonies emerged between day 6 and 10 which were then characterized by the expression of numerous spermatogonial and pluripotency related markers. After 15 days of subculture, the relative mRNA expression study revealed that 40 ng/mL concentration of Glial cell line-derived neurotrophic factor (GDNF) upregulated the expression of BCL6B, ID4, PLZF, and UCHL1. Moreover, the supplementation of GDNF + bFGF up-regulated the expression of PLZF and BCL6B. UCHL1 expression was higher after addition of GDNF + LIF while, THY1 overexpressed in response to the addition of GDNF + CSF1. These results demonstrated that the goat SSCs were efficiently propagated using a KOSR based serum-free media and the growth factor supplementation markedly influences their gene expression profile.
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Acknowledgements
The present work was funded by National Agriculture Innovation Project (NAIP) Grant to M.S.C. (C-2067 and 075) and S.K.S. (C 2–1-(5)/2007). The authors also acknowledge ICMR for providing Ph.D. fellowships to Ankur Sharma.
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SKS, RSM and MSC conceived the study. MR, DD, MT and MKS carried out the experiments. AS and PP wrote the paper. All authors have read and approved the final manuscript.
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Sharma, A., Shah, S.M., Tiwari, M. et al. Propagation of goat putative spermatogonial stem cells under growth factors defined serum-free culture conditions. Cytotechnology 72, 489–497 (2020). https://doi.org/10.1007/s10616-020-00386-8
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DOI: https://doi.org/10.1007/s10616-020-00386-8