胶原酶灌注法制备小鼠肝细胞

On the production of mouse hepatocytes by perfusion with collagenase

目的:建立体外培养小鼠肝细胞的实验体系。方法:采用两步原位胶原酶灌注法分离小鼠肝细胞,在RPM I1640培养液中培养。结果:用胶原酶灌注法分离的小鼠肝细胞活率可达90%以上,产量平均为2.8×107,完全满足实验要求。肝细胞在RPM I1640培养液中2 h即可贴壁,48 h内细胞贴壁充分,伸展良好,1周后死细胞增多。结论:胶原酶灌注法成功制备出小鼠肝细胞。

Objective： To establish experimental system of mouse hepatocytes culture in vitro. Methods： Mouse hepatocytes were isolated by a two-step perfusion with collagenase in situ , and cultured in RPMI1640. Results： The cells viability（ 90% ） and yield（ × 107） could meet the requirement of experiment. The cells attachment was noted after 2 h incubation in RPMI1640. During the following 48h, the cells attached completely, and extended well, but the death cells increased one week later. Conclusion Mouse hepatocytes can be prepared by perfusion with collagenase successfully.