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.

中文翻译：

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在生长因子下山羊假定的精原干细胞的繁殖定义了无血清培养条件。

在本研究中，我们使用无血清培养基繁殖山羊推定的精原干细胞（SSCs），并评估了关键生长因子对某些SSC标记和自我更新相关基因的相对表达的影响。将富集的SSCs在补充有10％KOSR的KO-DMEM中的同源Sertoli细胞饲养层上培养。推定的SSC菌落在第6天到第10天之间出现，然后以许多精原和多能性相关标志物的表达为特征。继代培养15天后，相对mRNA表达研究显示40 ng / mL浓度的胶质细胞源性神经营养因子（GDNF）上调了BCL6B，ID4，PLZF和UCHL1的表达。此外，GDNF + bFGF的补充上调了PLZF和BCL6B的表达。加入GDNF + LIF后UCHL1表达较高，而加入GDNF + CSF1则THY1过表达。这些结果表明，使用基于KOSR的无血清培养基可以有效地繁殖山羊SSC，并且添加生长因子会显着影响其基因表达谱。