体外不同诱导条件下对大鼠胚胎肝干细胞分化影响的实验研究  

Effect of different inducing conditions on rat fetal liver stem cells differentiation in vitro

作　　者：高植泉[1] 陶开山[1] 李韧[1] 尤楠[1] 张明[1] 季茹[1] 张福琴[1] 窦科峰[1] 

机构地区：[1]第四军医大学西京医院肝胆胰脾外科,陕西西安710032

出　　处：《胃肠病学和肝病学杂志》2010年第6期513-516,共4页Chinese Journal of Gastroenterology and Hepatology

基　　金：国家自然科学基金(30772094)

摘　　要：目的探讨和比较不同体外条件对大鼠胚胎肝干细胞分化成熟的影响。方法将取自孕14d胎龄F344大鼠胚胎肝组织的肝干细胞分别接种至含10、20、40ng/mLHGF,0.5%、1%、2%二甲基亚砜(DMSO),10ng/mLHGF+0.5%DMSO、20ng/mLHGF+1%DMSO、40ng/mLHGF+2%DMSO以及空白的培养基中进行体外培养15d;诱导15d后,ELISA检测和比较不同组细胞内甲胎蛋白(AFP)和白蛋白(ALB)的浓度,实时定量PCR比较不同组细胞ALB、葡萄糖6-磷酸酶(G-6p)、角蛋白8、18(CK-8、CK-18)的mRNA的表达水平。结果与空白对照组相比,诱导15d后各实验组肝干细胞ELISA检测AFP明显下降(P<0.01),ALB明显升高(P<0.01);实时定量PCR结果表明各实验组ALB、G-6P、CK-8、CK-18mRNA水平较对照组均明显升高,其中大鼠胎肝干细胞体外最佳诱导条件为20ng/mLHGF+1%DMSO(P<0.05)。结论 HGF和DMSO均可在体外诱导大鼠肝干细胞分化,不同浓度HGF和DMSO影响其分化成熟程度;HGF、DMSO对大鼠胚胎肝干细胞的分化具有协同效应。Objective To explore and compare the effect of different inducing conditions on differentiation of rat fetal liver stem cells in vitro.Methods Fetal liver stem cells were isolated at embryonal day（ED）14 from pregnant F344 rats.The cells were cultured in different mediums containing 10 ng/mL HGF,20 ng/mL HGF,40 ng/mL HGF,0.5% dimethyl sulfoxide（DMSO）,1% DMSO,2% DMSO,10 ng/mL HGF＋0.5% DMSO,20 ng/mL HGF＋1% DMSO,40 ng/mL HGF＋2% DMSO and the control medium respectively for 15 days.On day 15,the expressions of albumin（ALB）,G-6P,keratin 8（CK-8）and keratin 18（CK-18）mRNA within cells cultured in different mediums were measured by real-time quantitative PCR.The concentration of AFP and ALB within cells cultured in different mediums were measured by ELISA test.Results On the day 15,the concentration of AFP protein in each experimental group decreased significantly than that in the control group（P0.01）,while the concentration of ALB protein increased significantly（P0.01）;the expressions of ALB,G-6P,CK-8 and CK-18 mRNA were significantly higher than those in the control group.20 ng/mL HGF＋1% DMSO was the best inducing condition on differentiation of rat fetal liver stem cells in vitro（P0.05）.Conclusion Both HGF and DMSO have concentration-depend effect of inducing the differentiation of fetal liver stem cells in vitro.Furthermore,HGF and DMSO can play a synergistic effect on the differentiation of rat fetal liver stem cell.

关 键 词：肝干细胞 肝细胞生长因子 二甲基亚砜 细胞分化 

分 类 号：Q813[生物学—生物工程]