Li Ting Ting, Geng Shuang Shuang, Xu Hui Yan, Luo Ao Lin, Zhao Peng Wei, Yang Huan, Liang Xing Wei, Lu Yang Qing, Yang Xiao Gan, Lu Ke Huan
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, College of Animal Science and Technology, Guangxi University, Nanning 530005, China.
J Vet Sci. 2020 Jan;21(1):e13. doi: 10.4142/jvs.2020.21.e13.
Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system ( < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, ( < 0.05), ( < 0.01) and ( < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⁺ subset in the cells cultured in the defined materials system. The expression levels of ( < 0.05) and ( < 0.01) were significantly increased, and the expression levels of ( < 0.01) and ( < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs .
目前,水牛精原干细胞(SSCs)的培养体系多种多样,其效果仍不明确。在本研究中,我们比较了含有未定义成分(胎牛血清)和定义明确成分(KnockOut血清替代物)的培养体系对水牛类SSC细胞培养的影响。在使用定义明确成分的培养体系中观察到的DDX4和UCHL1阳性细胞(传代2代培养2天)显著多于使用未定义成分的培养体系(P<0.01),并且这些细胞维持的时间比使用未定义成分培养体系中的细胞更长(10天对6天)。此外,[具体基因名称1](P<0.05)、[具体基因名称2](P<0.01)和[具体基因名称3](P<0.05)在使用定义明确成分的培养体系中的表达水平显著高于使用未定义成分的培养体系。使用视黄酸诱导来验证培养的细胞是否保持SSC特征,结果显示在使用定义明确成分培养体系中培养的细胞中有一个SCP3⁺亚群。[具体基因名称4](P<0.05)和[具体基因名称5](P<0.01)的表达水平显著升高,而[具体基因名称6](P<0.01)和[具体基因名称7](P<0.05)的表达水平显著降低。这些发现为探索水牛SSCs的机制提供了更清晰的研究平台。