摘要
目的:探索原代培养大鼠肝细胞的最佳分离方法。方法:对比观察不同的肝细胞分离技术的培养效果。培养前,以4％台盼蓝染色判定肝细胞活性,培养48h后,观察肝细胞贴壁及生长状况。结果:①灌流消化法优于剪切消化法;②在多种灌流消化法中,经门静脉灌流消化法优于经胆总管灌流消化法及经腹主动脉灌流消化法;③在门静脉灌流的基础上,0.1％胶原酶Ⅰ37℃热消化10～15分钟,效果最好,而0.25％胰蛋白酶消化10～12分钟次之,用含EDTA的D-Hank液直接灌流分离法效果极差。结论:原代培养肝细胞的得率及活性与不同分离法包括灌流途径、灌流液、消化时间等有关。
Aim: To explore the best isolation methods of rat liver cells from primary culture. Method: The cultural effects of variant techniques of isolating rat liver cells were compared and observed. Before culture, the viability of liver cells was determined by 4% trypan blue after culture for 48 hours the adhesion and growth status of liver cells were observed. Result: ①Isolation digestion is superior to mechanical digestion by means of scissoring and cutting.②Among various kinds of isolation digestion technique , isolation by way of portal vein is superior to that by way of the common bile duct and abdominal aorta. ③On the basis of isolation by way of portal vein,under 37 C hot digestion for 10-15 minutes, 0. 1 % collagenase I is superior to 0. 25% trypsin. Action of immediate isolation by means of D-Hank's solution containing EDTA is the worst in all digestion methods. Conclusion:The survival rate and viability of liver cells from primary culture relate to iso lation methods including perfusion way, perfusate and digestion time etc. .
出处
《中西医结合肝病杂志》
CAS
1999年第4期19-21,共3页
Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases
基金
国家自然科学基金课题资助(批准号:39670910)
