成年大鼠肝细胞的分离及培养  

Isolation and cultivation of adult rat liver cells

作　　者：洪文清[1] 盛和章[1] 

机构地区：[1]军事医学科学院毒物药物研究所

出　　处：《军事医学科学院院刊》1993年第1期57-60,共4页Bulletin of the Academy of Military Medical Sciences

摘　　要：用含有0.5mmol/L EGTA的Hanks液100ml灌流大鼠肝脏,然后用100U/ml胶原酶溶液100ml循环灌流10min,再用100ml的0.5%胰酶灌流。分离的肝细胞用台盼蓝排斥试验显示87%～89%活率,产率可达4.7×10~8,具有~3H-TdR掺入活性。原代培养3d出现新增殖的肝细胞,20d增殖约5倍,可进行连续传代培养。A reproducible method for the isolation and cultivation of adult rat liver cells isdescribed. Under physiologic condition the liver cells of adult rats were dissociated aftercollagenase-trypsin perfusion. This method yielded a population of cells with a viability of87%-89%. the total number of viable cells might attain 4.7×10~8. Approximately 50% viable cellshad flattened out and attached to the plate surface after 24 hours of cultivation in Ham's F-12medium. Primary culture showed areas of dividing cells after 3 days in cultivation. In subsequentsubcultures, the dividing cells could be grown continuously. The rat liver cells showed normalDNA synthetic activities by HU inhibiting incorporation of ~3H-TdR to DNA and normal incor-poration of ~3H-TdR to DNA assay.

关 键 词：肝细胞 原代培养 胶原酶 胰酶 

分 类 号：R329-33[医药卫生—人体解剖和组织胚胎学]