猪雄性生殖干细胞的分离培养及鉴定

Abstract

Abstract: This study was aimed to explore the suitable conditions for the isolation,culture of pig male germline stem cells, and establish the in vitro culture system. The cells were isolated from the neonatal pig test by the two step enzyme digestion method, and cultured for further identification, compared the enrichment efficiency of cells on laminin and gelatin due to the different adherence velocity. And then the cells were identified for the alkaline phosphatase activity and the expression of the stem cell marker protein OCT-4. The results showed that laminin was more suitable for the enrichment and growth of porcine germline stem cells. The enrichment efficiency and proliferation rate of cells using laminin method was obviously superior to which using gelatin sorting. The cultured mGSCs had the similar morphology and proliferation characteristics to mGSCs of mouse. The expression of OCT-4 was positive in target cells, while it was negative in feeder cells-sertoli cells. The target cells showed strong expression of alkaline phosphatase, while nothing was detected in feeder cells-sertoli cells. The results showed that, we have established the preliminary culture system of porcine germline stem cells, with well maintained stem cell activity, normal replication function and differentiation potentials.

Key words: stem cell; germline stem cells; porcine; cell culture

CLC Number:

S828.43

KOU Xiao-qin, LIU Peng, LI Ying, YAN Hua, TANG Zhong-lin, MU Yu-lian, YANG Shu-lin, ZHOU Rong, AO Hong, LI Kui. Isolation, Culture and Identification of Porcine Male Germline Stem Cells[J]. , 2013, 40(1): 117-122.

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Isolation, Culture and Identification of Porcine Male Germline Stem Cells

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