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

Journal of Veterinary Science 2020년 21권 1호 p.13 ~ 13
 ( Li Ting-Ting ) - Guangxi University College of Animal Science and Technology

 ( Geng Shuang-Shuang ) - Guangxi University College of Animal Science and Technology
 ( Xu Hui-Yan ) - Guangxi University College of Animal Science and Technology
 ( Luo Ao-Lin ) - Guangxi University College of Animal Science and Technology
 ( Zhao Peng-Wei ) - Guangxi University College of Animal Science and Technology
 ( Yang Huan ) - Guangxi University College of Animal Science and Technology
 ( Liang Xing-Wei ) - Guangxi University College of Animal Science and Technology
 ( Lu Yang-Qing ) - Guangxi University College of Animal Science and Technology
 ( Yang Xiao-Gan ) - Guangxi University College of Animal Science and Technology
 ( Lu Ke-Huan ) - Guangxi University College of Animal Science and Technology

Abstract


Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro 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 in vitro 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 (p < 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, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 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 Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.

키워드

Buffalo; spermatogonial stem cells; primary cell culture
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