青海湖裸鲤体外肝胰细胞原代与传代培养

采用组织块移植培养技术，分别用DMEM和RPM11640培养基对青海湖裸鲤肝胰组织细胞进行原代培养。培养48h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。确立青海湖裸鲤肝胰细胞培养条件为：培养基为DMEM，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和添加细胞生长因子。

The primary culture and subculture of hepatopancreas cells from Qinghai lake-naked carp(Gymnocypris przewalskii) in vitro

By using the tissue explantation technique, primary culture of hepatopancreas tissue cells derived from naked carp （ Crymnocypris przewalskii） of Qinghai lake was experimented by using DMEM and RPMI1640 medium separately. The cells migrate out around tissues after 48 hours＇ culturing and proliferate into growth halo. The cells outgrow to monolayer about a week later. The experiment on subculture for monolayer cells was carried out and the 4^th generation cells were obtained after primary cells were digested with trypsase-EDTA solution. The optimized culture conditions for naked carp hepatopancreas cells are ： DMEM medium, 27 ℃, pH 7.0 - 7.5,20% calf serum on primary culture, 10% calf serum on subculture without CO2 and growth factor.