高原环境猪肝细胞体外球形聚集体培养

Cultivation of porcine hepatocyte spheroids at plateau in vitro

目的了解在高原环境条件下体外球形聚集体培养猪肝细胞的适宜接种浓度及其生长、增殖和生物学功能情况,为构建高原体外生物型人工肝脏寻找更加理想的生物材料。方法采用已适应本地环境的长白仔猪分离肝细胞,分别以浓度2×105/mL接种进行体外单层培养和以浓度1×106/mL、5×106/mL接种进行体外球形聚集体培养,对比观察培养细胞数量及培养上清液中总蛋白、白蛋白和尿素水平变化。结果接种4d后,3组细胞数量均明显增多,培养上清液中总蛋白、白蛋白和尿素水平均呈现显著升高;球形聚集体培养显著高于单层培养,并且5×106/mL接种组更高。结论浓度5×106/mL接种的球形聚集体培养猪肝细胞的生长、增殖及生物学功能更好,用作构建高原体外生物型人工肝脏的生物材料可能更加理想。

Objective To understand the characters of growth,proliferation,and biological function of the porcine hepatocytes cultured in the environment of plateau,and find more ideal biological material for constructing plateau bioartifical liver in vitro.Methods The hepatocytes isolated from the porcine liver were cultivated in monolayer（2 × 105/mL） and spheroid（1 × 106/mL,5 ×106/mL）,respectively.The cell amounts and the biochemical substance levels in culture supernatant of the cultured porcine hepatocytes were investigated at the different time.Results After cultured 4 days,the amounts of cultured porcine hepatocytes increased in 3 groups,and the levels of total protein,albumin and urea in culture supernatants increased evidently.The cell amounts and the biochemical substance levels of spheroid group（5 × 106/mL） were higher than that of monolayer group in the same unit culture fluid.Conclusion The growth,proliferation and biological function of the cultured porcine hepatocyte spheroid（5 ×106/mL） is appropriate for establishing plateau bioartifical liver and the porcine hepatocytes cultured in spheroid may be better than that of in monolayer.