To enrich bovine gonocytes from cryopreserved testicular tissues, the cryoprotection effects of the freezing media containing knockout serum replacement (KSR) were examined. Using Minimum essential medium (MEM) + 10% dimethyl sulfoxide (Me2SO) as the basic medium, calf testicular tissues were cryopreserved in media containing 0, 5, 10, 20, 40, 90% KSR and 5% fetal bovine serum (FBS) respectively. Morphologically the seminiferous cords and interstitium were well preserved in all groups. The gonocytes were all glial cell line-derived neurotrophic factor (GDNF) family receptor α-1 (GFRα-1) positive. The recovery rates in all KSR groups were higher than that of the 10% Me2SO group, while comparable to the 5% FBS group. The enriched gonocytes expressed gonocyte marker GFRα-1 typically. Collectively supplementation of 5-10% KSR can achieve comparable cryoprotective effects with using 5% FBS, which is useful in future study due to its defined formulation that is more consistent in quality and stable in supply.

中文翻译：

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用敲除血清替代物冷冻保存小牛睾丸组织

为了从冷冻保存的睾丸组织中富集牛生殖细胞，研究了含有敲除血清替代品 (KSR) 的冷冻培养基的冷冻保护作用。以最低必需培养基（MEM）+10%二甲亚砜（Me2SO）为基础培养基，将小牛睾丸组织冷冻保存在含有0、5、10、20、40、90% KSR和5%胎牛血清（FBS）的培养基中分别。在形态上，所有组的生精索和间质都保存完好。生殖细胞均为神经胶质细胞源性神经营养因子（GDNF）家族受体α-1（GFRα-1）阳性。所有 KSR 组的回收率均高于 10% Me2SO 组，而与 5% FBS 组相当。富集的生殖细胞通常表达生殖细胞标志物 GFRα-1。