5-氮胞苷诱导心肌组织中c-kit~+干细胞分化的实验研究

目的:创建小鼠心肌干细胞(cardiac stem cells,CSC)分离、培养的方法,借助干细胞表面标记c-kit纯化CSC,并用5-氮胞苷诱导CSC分化为心肌细胞。方法:用酶定时消化方法分离提取小鼠心脏组织中的细胞,进行体外培养;免疫磁珠法纯化c-kit阳性细胞,并用流式细胞仪检测纯化后细胞表面标记c-kit、lin和CD34的表达;以免疫细胞化学染色法检测诱导后心肌特异性蛋白T、连接蛋白-43、心肌细胞特异性结蛋白;结合形态学变化,分析不同诱导条件作用结果之间的差异。结果:从C57BL小鼠心肌组织中分离消化所得的细胞中,含有一种小而圆形、折光性强的细胞,经传代培养的细胞增殖速度比原代细胞明显为快,存在较多的类似于干细胞增殖和形态学特点的细胞。用免疫磁珠法可以分选出高纯度的c-kit+CSC,这种CSC基本不表达CD34和lin。在不同诱导条件下,此种CSC可以分化为搏动的细胞,并能表达心肌特异性蛋白。结论:从C57BL小鼠心肌组织中可以获得CSC,用免疫磁珠法可以分选出表型为c-kit+-CD34--lin-高纯度的CSC,经5-氮胞苷诱导或改变CSC生长的微环境后,此种细胞可以分化成心肌细胞,且能形...

Empirical study on c-kit cardiac stem cells induced differentiation into cardiomyocytes by 5-aza

AIM: To find the method of dissociating and culturing the cardiac stem cells(CSC),purify CSC by cell surface markers of stem cell with c-kit,and induce CSC differentiation into cardiomyocytes by 5-azacitidine(5-aza) in vitro.METHODS: The heart cells were dissociated and extracted by zymo-timing digestion and cultured in vitro,the c-kit masculine cells as CSC markers were purified by immunomagnetic beads method.The expression such as c-kit,lin and CD34 as CSC markers were detected after purification by flow ...