青海湖裸鲤肾细胞的原代培养和传代培养

采用组织块移植培养技术,用RPMI 1640培养基对来源于青海湖裸鲤(Gymnocypris przewalskii)肾组织的细胞进行原代培养。结果显示:培养48 h可见组织块周围有细胞迁出,并形成生长晕;培养一周可形成单层细胞;对原代培养的单层细胞用胰蛋白酶-EDTA消化后,传代培养至第四代。实验初步确立青海湖裸鲤肾细胞培养条件为:培养基RPMI 1640,培养温度为27℃,pH值为7.0~7.5,原代培养血清浓度为20%,传代培养的血清浓度为10%,无需通入CO2。

The Primary Culture and Subculture of Renal Cells from Naked Carp of Qinghai Lake(Gymnocypris przewalskii)

By using the tissue explantation technique,primary culture of renal tissue cells derived from naked carp of Qinghai lake(Gymnocypris przewalskii)were experimented by using RPMI 1640 medium.The cells migrated out around tissues after 48 hours' culturing and the growth halo emerged.Under this condition,monolayers of primarily cultured renal cells were routinely obtained within a week.The 4th generations cells were obtained after subculture of primary cells with trypsin-EDTA digestion method.The optimized conditions for naked carp renal cells are:medium RPMI 1640,27 ℃,pH 7.0~7.5,20% bovine calf serum on primary culture,10% bovine calf serum on subculture,no CO2.