三种细胞培养模型的效果评价

Effect evaluation of three cell culture models

原代大鼠肝细胞经不同体外细胞培养模型培养后 ,对肝细胞的酶渗出量、白蛋白分泌水平和细胞色素P450 1A(CYP 1A)活性进行了分析。结果显示 :在三种培养模型中 ,培养液的LDH水平随培养时间的延长而逐渐降低 ,但在单层培养 (MC)的第 5天 ,LDH水平明显升高 ,而夹层培养 (SC)的LDH在第 8天后没有检出 ,AST和ALT水平在整个培养过程中无明显变化。在生物反应器中 (bioreactor) ,肝细胞基础CYP1A活性可维持 2周以上 ,培养液中白蛋白的含量以生物反应器最高 ,其次是SC和MC。随着培养时间的延长 ,MC和SC中肝细胞CYP 1A活性逐渐降低 ,且MC肝细胞CYP 1A活性的下降速度比SC快。这种现象可被细胞色素P450 (P450 )的诱导剂如奥美拉唑和 3 甲基胆蒽 (3 MC)部分改变 ,MC肝细胞的诱导作用总是比SC肝细胞强。结果证实每一个体外细胞培养模型都有各自的优缺点 。

Primary rat hepatocytes were cultured using three kinds of models in v itro and the enzyme leakage, albumin secretion, and cytochrome P450 1A (CYP 1A) activity were observed. The results showed that the level of LDH in the medium d ecreased over time in the period of culture. However, on 5 days, LDH showed a si gnificant increase in monolayer culture (MC) while after 8 days LDH was not dete cted in sandwich culture (SC). The levels of AST and ALT in the medium did not c hange significantly over the investigated time. The basic CYP 1A activity gradua lly decreased with time in MC and SC. The decline of CYP 1A in rat hepatocytes w as faster in MC than that in SC. This effect was partially reversed by using cyt ochrome P450 (CYP450) inducers such as omeprazol and 3-methylcholanthrene (3-M C) and the CYP 1A induction was always higher in MC than that in SC. Basic CYP 1A activity in bioreactor was keeped over 2 weeks and the highest albumin productio n was observed in bioreactor, and next were SC and MC. In conclusion, our result s clearly indicated that there have some advantages and disadvantages in each of models in which can address different questions in metabolism of toxicants and durgs.