【摘要】 PLC/PRF/5肝癌细胞系(PLC)经适应于本实验室含10％小牛血清的Eagles’营养液(csEagles’液),传代3天后即可长成单层。应用固相放射免疫测定法(SPRIA)动态检测产生乙型肝炎表面抗原(HBsAg)量,发观随培养天数延长,HBsAg产量增多,到18天左右达高峰。与纯化HBsAg此较,最高产量(14～18天)约为200ng/ml/10~5细胞。HBsAg产量的增多与PLC细胞出现溶酶体及线粒体退变、细胞融合及脱落死亡圆细胞数增多呈平行关系。这些现象与认为产生HBsAg是PLC生长周期末期活动的看法相符。自营养液中去除小牛血清,则细胞仅能短期维持,第4天起开始脱落,但4天内对HBsAg产量无显著影响。加入黄芪多糖培养,未能延长细胞生长寿命或明显提高HBsAg产量。更多还原

【Abstract】 PLO/PRF/5 hepatoma cell line (PLO) adapted to Eagles’ medium supplemented with 10% of inactivated calf serum in our laboratory was successfully passaged, and confluent growth was generally observed on the third day after seeding. The production of hepatitis B surface antigen(HBsAg)in tissue culture fluid was serially determined by solid phase radioimmunoassay. Prolonged incubation of culture in growth or maintenance media resulted in a progressive increase in HBsAg, which was maximum around 18 days, and then declined gradually. Compared with highly purified HBsAg prepared from sera of carriers, maximum production of HBsAg from PLO cultures collected between 14-18 days was about 200ng/ml/105 cells. The increase of HBsAg production was paralleled by cytological changes, in which there were degenerations in lysosomes and mitochondria, fusion of cells, and increase in the number of detached and non-viable rounded cells. These were in agreement with the suggestion that HBsAg production is cell cycle related to and representing a terminal event. Depletion of calf serum from media within four days of culture had no significant effect on the production of HBsAg, but after four days cells were found to be detached from flaska, followed by decreased production of HBsAg. When polysaooharides of Eadix Astragali Seu Hedysari (BAH), a Chinese herb which has been shown to prolong the life-span of human embryo lung diploid cell strain was added to culture media, the life-span of PLC, however, was not prolonged, nor was the production of HBsAg enhanced significantly.更多还原