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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。
The PLC / PRF / 5 hepatoma cell line (PLC) was adapted to our laboratory with 10% calf serum in Eagles ’juice (csEagles’ solution) and grew into a monolayer after 3 days of passage. The amount of HBsAg produced by dynamic detection of SPRIA was measured. The amount of HBsAg produced increased with the prolongation of culture days and reached the peak at 18 days. Compared with the purified HBsAg, the highest yield (14-18 days) was about 200ng / ml / 10 ~ 5 cells. HBsAg production increased with PLC cells lysosomal and mitochondrial degeneration, cell fusion and exfoliated death increased in parallel with the number of round cells. These phenomena are consistent with the belief that HBsAg production is the end of the PLC growth cycle. When bovine calf serum was removed from nutrient solution, the cells were only maintained for a short period of time and began to fall off on day 4, but had no significant effect on HBsAg production within 4 days. Astragalus polysaccharides added to the culture, failed to extend the life of cell growth or significantly improve HBsAg yield.