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目的:通过对SD乳鼠体外自发性搏动的心肌细胞(CMs)分离培养,并观察其超微结构,以了解心肌干细胞的特性。方法:用体外分离、纯化培养SD乳鼠自发性搏动的心肌细胞,应用倒置显微镜及透射电镜观察其形态结构与超微结构,并进行心肌特异性标志troponin I(cTnI)的免疫细胞化学鉴定。结果:体外分离、纯化培养出心肌细胞,细胞具有自发性搏动特性,表达心肌特异性抗体Tropnin I。电镜下可见细胞内出现肌丝样结构及Z线结构,细胞普遍存在肌小节,细胞间有闰盘连接;部分细胞表面有大量微绒毛,细胞器丰富,可见内质网、线粒体。结论:体外可分离培养扩增出自律性搏动的CMs,这些细胞表现增殖活跃,具有干细胞的增殖旺盛特性。
OBJECTIVE: To isolate and culture spontaneously pulsatile cardiomyocytes (CMs) of neonatal SD rats and to observe its ultrastructure to understand the characteristics of cardiac stem cells. Methods: Spontaneous beating cardiomyocytes were isolated and purified from SD neonatal rats. The morphology and ultrastructure were observed by inverted microscope and transmission electron microscope. Immunocytochemistry was used to identify cardiac troponin I (cTnI). Results: Myocardial cells were isolated and purified in vitro. The cells had spontaneous pulsatile characteristics and expressed cardiac specific antibody Tropnin I. Electron microscopy showed myofilament-like structure and Z-line structure in the cells. The cells were common in muscle sections and intercalated with intercalated discs. Some of the cells had a large number of microvilli on the surface, with rich organelles and endoplasmic reticulum and mitochondria. CONCLUSION: Self-paced CMs can be isolated and cultured in vitro. These cells show proliferative activity and proliferative characteristics of stem cells.