肝脏部分切除对骨髓间充质干细胞定向肝细胞分化的影响

Effect of partial hepatectomy to the rate of differentiation from bone marrow musenchymal stem cells into hepatocytes in vitro

目的:探讨肝脏损伤对小鼠骨髓间充质干细胞(MSCs)向肝细胞诱导分化的影响。方法:昆明系小鼠分两组,分别行肝脏2/3切除和开关腹手术,分别于术后12 h,24 h,36 h,48 h,72 h后提取MSCs,取第3代的MSCs,流式细胞仪鉴定MSCs后,应用HGF联合EGF诱导,20 d后免疫荧光染色鉴定肝细胞标志物ALB,CK8,计数ALB阳性细胞数量和同视野细胞总数,计算阳性率。结果:诱导分化20 d,两组均有ALB、CK8表达,24 h时间点,实验组阳性率(10.41±0.11)%,对照组阳性率为(9.83±0.32)%,实验组与对照组阳性率比较差异有显著性意义;其余时间点实验组和对照组阳性率比较差异均无显著性意义(P>0.05)。结论:肝脏损伤后24 h,提取MSCs定向肝细胞的阳性率高。

Objective：To investigate the effect of partial hepatectomy to mesenchymal stem cell （MSCs）differentiating into hepatocytes. Methods：The mice were randomly separated into two groups （n = 20）, one group was operated partial hepatectomy, the other group as control. To extract the MSCs at the time of 12 h, 24 h, 36 h, 48 h, 72 h after operation respectively. MSCs of the third generation was cultured by IMDM containing hepatocyte growth factor（HGF） and epidermal growth factor （EGF） for 20 days. The expressions of ALB and CK8 were detected with immunofluorescence; Counting the ALB positive cell and calculating the positive rate. Results： After 20 days inducing, the differentiated cells expressed ALB and CK8. The number of positive ceils in experimental, group is more than control, especially at 24h after operation. The positive rate of experimental group is（ 10.41 ±0.11 ）%, the control group is （9.83 ±0.32）%, the positive rate of the experimental group achieved statistical significance com- pared with the control （P 〈 0.05） at 24h after operation. Conclusion： The positive rate from MSCs into hepatocytes was improved at 24h after operation.