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采用胰酶半消化法,对贝氏高原鳅的吻端、肾脏和性腺组织的原代培养条件进行探究,使用MEM,RPMI1640,DMEM和DMEM/F12培养基进行培养;血清质量分数设置为10%,30%,50%和100%;分别置于37℃,26℃和室温(11～15℃)条件下培养,观察细胞在不同条件下的生长情况.初步确定了各组织原代培养的最佳条件是温度26℃、血清质量分数50%的条件下,吻端和肾脏组织在MEM培养基、精巢在DMEM培养基中生长情况最优.Giemsa染色显示各组织细胞结构完整,荧光染料Alexa fluor 488Phalloidin显示各组织细胞的微丝网络发达.
The primary culture conditions of the snout, kidney and gonad tissues of Misgurnus anguillarum were investigated by trypsin digestion, and cultured in MEM, RPMI1640, DMEM and DMEM / F12 medium. The serum concentration was set at 10% , 30%, 50% and 100%, respectively. The cells were cultured under the conditions of 37 ℃, 26 ℃ and room temperature (11-15 ℃), and the growth of the cells under different conditions was observed. The optimal conditions were that the temperature and the serum concentration were 26 ℃ and 50%, respectively, and the growth of the testis and kidney tissue was best in the MEM medium and the testis in the DMEM medium.The results of Giemsa staining showed that the cell structure of each tissue was intact. The fluorescent dye Alexa fluor 488Phalloidin shows that the microfilament network of each tissue cell is well developed.