旋转生物反应器内微载体大规模扩增肝细胞

Rapidly large scale proliferation of liver cell line by microcarrier vehicle in rotary cell culture system

目的评估旋转生物反应器(RCCS)内肝细胞微载体大规模培养的可行性。方法为获取足够数量活性良好的肝细胞种子细胞,本实验探索在RCCS内应用微载体技术快速扩增肝细胞的方法。将CL-1细胞和HepFMMU应用Cytodex-3微载体在RCCS内进行动态培养,观测肝细胞的生长情况和细胞代谢率。结果肝细胞在Cytodex-3微载体上生长迅速、生长倍增时间缩短,并保持很高的生物活性。培养至第5天,细胞数量可达最初接种的23倍。细胞的数量在第5天左右达到最高,然后开始下降。功能学检查、倒置显微镜、扫描电镜及MTT均提示细胞功能良好。结论RCCS微载体肝细胞大规模培养是一种可行的细胞快速扩增方法,但对于人工肝庞大的细胞数量和功能需求,应进一步研究,通过优化营养和废物排除,模拟肝脏生理结构,提高细胞培养规模和功能。

Objective To evaluate the feasibility of large scale culture of liver cell by microcarrier vehicle in rotary cell culture system （RCCS）. Methods Liver cell line CL-1 and HepFMMU were rapidly proliferated by mierocarrier vehicle in RCCS in a short period of time. Histomorphology and biological functions were evaluated accordingly in phase contrast observation, scanning electron microscope （ SEM）, biological functions test and MTT test. Results Liver cell cultivated in the bioreactor in three dimensions were expanded rapidly with shorter doubling time. The cell population could reach 23 times more than that before expansion in fifth day and decreased afterward. The expanded ceils performed well in major functions examined by inverted microscope in phase contrast observation, scanning electron microscope （SEM） , biological functions test and MTT test. Conclusion It is feasible for rapidly large scale amplification for liver cell by mierocarrier vehicle in RCCS. But further research is needed to improve the supply of energy and oxygen and enhance the clearness of waste of RCCS though better simulating the liver structure due to high demand of liver cell density.