论文部分内容阅读
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。
Objective To explore the reasonable conditions for isolating hepatic parenchymal cells and non-parenchymal cells. Methods The collagenase / proteinase E digestion and low speed centrifugation were used to detect the cell quality by the cell viability, cell yield and cell purity. Results The hepatic parenchymal cell yield (98 × 10 9 ± 4 × 10 9 · L -1) and the purity (97%) were significantly higher under the conditions of 37 ℃, 0 5 g · L -1 collagenase for 30 min and 80 0 r · min -1 centrifugation. % ± 2%) and viability (> 90%) were the best. Collagenase and proteinase E combined, the highest non-parenchymal cell yield (286 × 10 9 ± 3 7 × 10 9 · L-1), Kupffer cell purity (16 0% ± 35%), and Kupffer cell content In line with the normal distribution of the body. Conclusion The separation of cells by this experimental method is of practical significance for the further study of liver parenchyma, especially non-parenchymal liver cells.