In this study, we investigated the effect of glial cell line-derived neurotrophic factor (GDNF), fibroblast growth factor (FGF) 2, and epidermal growth factor (EGF) on the expression of some self-renewal-related microRNAs (miRs) in putative buffalo spermatogonial stem cells (SSCs). The SSCs were cultured on a buffalo Sertoli cell feeder layer, colony formation was observed between 7 and 10 days. The SSC colonies expressed markers specific for undifferentiated type A spermatogonia and pluripotency markers. After 15 days of initial culture, the colonies were subcultured as treatment (supplemented with 20 ng mL-1 GDNF +10 ng mL-1 FGF2 + 10 ng mL-1 EGF) and control groups. The number and area of SSC colonies were significantly (p < 0.05) higher in the treatment group than in the control group. The relative abundance of miR-20b, miR-21, and miR-106a in SSCs supplemented with growth factors was significantly higher (p < 0.001) than that in the control. The results indicate that supplementation of SSC culture medium with growth factors (GDNF, FGF2, and EGF) may promote the expression of miR-20b, miR-21, and miR-106a, which is essential for self-renewal and maintenance of SSCs.

中文翻译：

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胶质细胞源性神经营养因子，成纤维细胞生长因子2和表皮生长因子的补充通过上调miR-20b，miR-21和miR-106a的表达促进假定的水牛（Bubalus bubalis）精原干细胞的自我更新。 。

在这项研究中，我们调查了神经胶质细胞源性神经营养因子（GDNF），成纤维细胞生长因子（FGF）2和表皮生长因子（EGF）对某些自我更新相关microRNA（miRs）表达的影响。推定的水牛精原干细胞（SSC）。SSCs在水牛Sertoli细胞饲养层上培养，在7至10天之间观察到菌落形成。SSC菌落表达了对未分化的A型精原细胞和多能性标记具有特异性的标记。初始培养15天后，将菌落传代培养（补充20 ng mL-1 GDNF +10 ng mL-1 FGF2 + 10 ng mL-1 EGF）和对照组。治疗组中SSC菌落的数量和面积显着高于对照组（p ＜0.05）。miR-20b，miR-21，补充有生长因子的SSC中的miR-106a和miR-106a明显高于对照组（p <0.001）。结果表明，向SSC培养基中添加生长因子（GDNF，FGF2和EGF）可能会促进miR-20b，miR-21和miR-106a的表达，这对于SSC的自我更新和维持至关重要。