人幼儿心肌细胞体外培养的实验研究

The study for culture of the human children heart muscle cells in vitro

目的探讨人幼儿心肌细胞体外培养、扩增以及可能用途。方法体外循环手术时获取少量1～5岁儿童心肌组织,制成细胞悬液用含10%胎牛血清的DMEM培养基进行体外培养、扩增,观察细胞形态变化,进行免疫组化鉴定并用电镜观察细胞超微结构。结果体外培养的幼儿心肌细胞至5、6代虽然仍具有心肌细胞超微结构,并可以被抗肌动蛋白、肌球蛋白、支架蛋白的单克隆抗体染色,但第4代以后细胞增殖速度变慢,细胞逐渐变形,第6代基本停止生长。结论人发育阶段的心肌细胞,在有限的体外培养过程中,数目可扩增,其亚显微结构和成分基本稳定,有可能作为心肌细胞移植的组织材料。

Objective To probe the condition for cultivation ,expansion of the human children heart muscle cells in vitro and possible uses. Methods Some heart muscle cells obtained from the children of 1-5 year old when the CPB operation had been performed were used to culture in vitro with DMEM medium （10% FBS）, used to analyse with immunohistochemistry, observe under electron microscopy. Results The developing children heart muscle cells for cultivation of passage 5,6 in vitro still can keep the ultrastructure of cardiomyoeytes. They also can be stained with anti-Actinin , anti-Myosin, anti-Desmin, but the cells began to hyperplasia slowly after passage 4 and change the shape gradually. The cells of passage 6 almost stopped to grow. Conclusion The developing children cardiomyocytes could be proliferated in culturing in vitro. It may be an ideal material of providing cells for the cellular cardiolnyoplasty.