摘要
目的体外分离、培养新生大鼠肝干细胞（HSCs）,并对其生物学特性进行鉴定。方法采用剪碎加酶消化法分离新生大鼠肝干细胞,用体积分数10%胎牛血清的H-DMEM培养液培养。以MTT法检测细胞的增殖情况,以RT-PCR检测肝干细胞特异性标记分子的表达。结果分离的新生大鼠肝干细胞体外培养24h已贴壁,3d左右可见克隆形成,呈铺路石样分布,光镜下细胞为方形或多角形,边缘清楚,细胞核及核仁明显,表达细胞角蛋白CK18、CK19、CK8、甲胎蛋白（AFP）等肝干细胞特异性标记分子,低度表达白蛋白。结论成功分离、鉴定出了新生大鼠肝干细胞,并可在体外条件下进行传代扩增培养。
Objective To isolate and cultivate hepatic stem cells （HSCs） from neonatal rat liver and identify their biological features.Methods HSCs from neonatal rat liver were isolated by mechanical cutting and enzyme digesting method,then cultivated in H-DMEM with 10% fetal bovine serum （FBS）. The cell proliferation was assayed by MTT test,and the specific biomarkers expression of HSCs was detected by RT-PCR.Results The isolated HSCs from neonatal rat liver adhered within 24 h,and cell clones were formed 3 d after cultivation. The cells presented slabstone-like,square or polygon appearance with distinct borderline under the light microscope. The RT-PCR results showed that the cells expressed CK18,CK19,AFP and ALB.Conclusion The cultivated cells from neonatal rat liver were proved to be HSCs and could proliferate in long term.
出处
《广东药学院学报》
CAS
2010年第4期416-419,共4页
Academic Journal of Guangdong College of Pharmacy
基金
国家自然科学基金资助项目(30973913)
国家科技重大专项资助项目(2009zx09103-407)
