体外自发性搏动乳鼠原代心肌细胞的分离培养及超微结构

Primary Spontaneously Beating Cardiac Cells from Neonatal Rats:Isolation,Culture and Ultrastructures

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, culture and study the ultrastructure of primary spontaneously beating cardiac cells from neonatal SD rat. nethods：Cardiomyocytes （ CMs ） derived from neonatal SD rats were isolated and purified in culture, and studied for morphology and ultrastructures by inversed microscope and transmission electron microscopy. Expression of cardiac-specific marker, troponin I （ cTnI）, was identified by immunohistochemistry. Results： CMs were successfully isolated and cultured in vitro, and displayed spontaneous beating features with positive staining for cTnl. Under transmission electron microscopy, myofilaments, Z bands, widespread sarcomeres and intercalated disks were seen. Some CMs had abundant micro-villi on their surface, plenty of endoplasmic reticula and mitochondria. Conclusion： CMs can be successfully cultured and expanded in vitro. The cultured cells appeared actively proliferating, which can feature stem cells.