肝小叶型生物反应器的初步研究

This study is to construct a novel bioreactor of extracorporeal bioartificial liver support system with woven hollow fibers and to improve the function of traditional hollow fiber bioreactor. For the purpose, firstly, a skill of hepatocytes isolate was improved in vitro and a simple extracorporeal perfusion apparatus of two-step perfusion was constructed. Secondly, a experimental method of hepatocytes isolated and purified by isopyknic centrifugation and selective attachment was established. Third, the spheroidal aggregate culture of human hepatocytes, collagen-gel immobilisation culture of rat hepatocytes and, sandwich-cultured by collagenous matrix of pig hepatocytes were succeed carried out. On these bases, a hollow fiber bioreactor with cultured pig hepatocytes was made up using an artificial capillary cell culture system. A novel 3-D network bioreactor of woven hollow fibers was developed and its function was examined. The results show that both the biological synthesize function and biological transformation function of the bioreactor and hepatocytes were maintained well. This bioreactor achieved to the design requirement, which play an importment base for the study of bioartificial liver with cultured hepatocytes.

采用体外两步灌流法消化肝组织，差速、密度梯度、淘析离心获取肝细胞、枯否细胞、窦内皮细胞，在氧合培养液循环装置辅助下，仿肝小叶细胞存在方式，于网织状中空纤维支架型反应器内进行肝细胞的混合三维立体培养，应用电镜、生化分析和转化代谢实验对肝小叶型反应器的生物功能及特征进行系统的评价，为体外人工肝系统和肝组织工程研究奠定基础。