大鼠肝实质及非实质细胞分离和质量检测

目的　探索分离肝实质细胞及非实质细胞的合理条件。方法　应用胶原酶 /蛋白酶E消化分离和低速离心技术 ,以细胞活率、细胞产量和细胞纯度等指标检测细胞质量。结果　在 37℃ ,0 5g·L-1的胶原酶作用 30min ,80 0r·min-1离心条件下 ,肝实质细胞产量 (98× 10 9± 4× 10 9·L-1)、纯度 (97%± 2 % )和活率 (>90 % )最好。胶原酶和蛋白酶E合用 ,肝非实质细胞产量最高 (2 8 6× 10 9± 3 7× 10 9·L-1) ,Kupffer细胞纯度合理 (16 0 %± 3 5 % ) ,且Kupffer细胞含量符合体内正常分布。结论　本实验方法分离细胞对于进一步研究肝实质 ,尤其肝非实质细胞功能有实际意义。

Isolation and characterization of hepatic parenchymal and nonparenchymal cells from rats

AIM To explore the appropriate separating condition of the parenchymal and nonparenchymal cells. METHODS The cell viability,yield and purity were measured by different separating methods. RESULTS Parenchymal cells were obtained with 800 r·min -1 and 37℃ for 40 min. Both the cell yield(98×10 9±3 5×10 9·L -1 ) and the cell purity(97%±2%) were higher;the cell viability was more than 90%. Compared with the other methods, nonparenchymal cells obtained with combined collagenasepronase methods had the highest cell yield(28 6×10 9±3 7×10 9·L -1 ), even the percentage of Kupffer(16%±3 5%) was reasonable and accorded with normal distribution. CONCLUSION These methods are very practical to detect the functions of above two types of cells, especially the nonparenchymal cells.