无血清培养高密度乳猪肝细胞的形态和功能变化

CHANGES IN MORPHOLOGY AND FUNCTIONS OF SUCKING PIG HEPATOCYTES CULTURED IN SERUM-FREE MEDIUM AT HIGH-DENSITY

本文研究了无血清培养高密度猪肝细胞的形态和功能变化。将分离的肝细胞以高密度(1×10~7/ml)培养在含激素、多种生长因子和营养成分的无血清培养基中,动态观察培养7天中肝细胞形态、活率、蛋白质合成功能、G-6-Pase活性、安定转化功能及LDH含量;同时以无血清培养低密度(5×10~5/ml)肝细胞作为对照组。研究结果表明:高密度培养的 肝细胞各项功能较低密度培养的肝细胞为低;高密度培养的肝细胞的形态、蛋白质合成功能在培养7天中保持稳定;活率随着培养时间的延长而下降,但均高于90％;安定转化功能在培养第2、3天最强;G-6-Pase活性在培养1天后明显下降,然后维持在较低水平;LDH含量在第1、2、3天较高。

In the present study, we assessed the dynamic changes in morphology and functions of porcine hepatocytes cultured in serum-free medium at high-density. Isolated hepatocytes were cultured in serum-free medium at high-density. Morphology, viability, protein synthesis, G-6-Pase activity, diazepam transformation of hepatocytes and LDH in supernatant was evaluated during 7 days of culture. The hepatocytes cultured in serum-free medium at low-density were served as a control group. The results showed that morphology, protein synthesis of hepatocytes cultured at high-density were stable over the course of 7 days. High viability ( >90% ) was obtained though it declined with time. Diazepam transformation was higher on days 2 and 3. G-6-Pase activity of the hepatocytes declined apparently after 1 day of culture, and it was maintained at low level for 7 days. LDH in supernatant was higher on days 1, 2 and 3. The functions of hepatocytes cultured at high-density were lower than at low-density culture.