新生大鼠心肌细胞的体外培养和生物学观察

Culture in vitro and Observation of Neonatal Rat Cardiac Myocytes

体外分离培养心肌细胞,为心肌组织工程提供种子细胞。采用胰酶连续消化法,分离新生大鼠心室肌细胞并在体外培养,倒置显微镜下观察细胞形态和搏动情况,并通过免疫组化染色法,检测心肌细胞的α肌节型肌动蛋白及连接蛋白Cx43。结果表明:培养2d后,心肌细胞开始自发搏动,在培养过程中心肌细胞的搏动频率逐渐加快,细胞聚集成团,随后成簇同步搏动,心肌细胞搏动可维持60d以上;α肌节型肌动蛋白及连接蛋白Cx43的表达量也逐渐增加。本研究体外培养出了状态良好的心肌细胞,在体外二维培养环境下,心肌细胞也有一定程度的生长发育,为心肌组织工程种子细胞来源的研究提供了实验依据。

To seperat and calture cardiac myocytes in vitro for cardiac tissue engineering.Cardiac myocytes from ventricles of neonatal rats were isolated by trypsin sequential digestion and cultured in vitro.The morphology and beating of cardiac myocytes were observed with phase-contrast microscopy and the α-sacromeric actin and connexin43 of the cells were detected with immunocytochemical staining.Result showed that about 2d after plantation,cardiomyocytes began to beat spontaneously and the beating frequency quickened,cardiomyocytes got together and beat synchronously in clusters gradually and the beating of cardiomyocytes lasted at least 60 days.The expression of the cells ′α-sacromeric actin and connexin43 were increased gradually.Cardiac myocytes in order state are isolated and cultured in vitro successfully and in 2-dimension culture environment,the cardiac myocytes grow and develop by degree.The study provide seed cells for cardiac tissue engineering.