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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 .

Effects of different culture systems on the culture of prepuberal buffalo (Bubalus bubalis) spermatogonial stem cell-like cells in vitro