【摘要】 目的建立体外培养东方田鼠鼠肝细胞的实验体系。方法采用下腔静脉插管原位灌注结合胶原酶灌注法分离纯化出肝细胞,在改良Eagle培养基(Dulbecco’s Modified Eagle Media,DMEM)中培养,显微镜下观察细胞形态,AO-EB染色观察测定其活率来评价细胞质量。结果一只东方田鼠肝脏一般可获得1.5×108～3.0×108个细胞,活率达95%,完全符合实验要求。刚分离的东方田鼠肝细胞呈圆球形,大小均匀,培养3h后,大部分肝细胞出现贴壁,形态呈扁平状。培养24h后,伸展良好,并成片生长。结论下腔静脉插管原位灌注结合胶原酶灌注法可以分离出高纯度和活率高的东方田鼠肝细胞。更多还原

【Abstract】 Objective To establish experimental system of Microtus fortis hepatocytes culture in vitro. Methods Microtus fortis hepatocytes were isolated and purified by inferior vena cava catheter perfusion with collagenase in situ, and cultured in Dulbecco’s Modified Eagle Media(DMEM) medium. Cells morphology was observed under an inverted microscope. Cells motility was determined by AO-EB staining. Results The cells yield (1.5×108～3.0×108) and viability(95%) could meet the requirement of experiment.The primary hepatocytes were round in shape,and the cells distributed uniformly. The cells attachment was noted after 3 h incubation,and most of the cells were flat.The cells formed sheet after 24 h cultivation and extended well. Conclusion High purity and viability Microtus fortis hepatocytes could be obtained by inferior vena cava catheter perfusion with collagenase in situ.更多还原