小鼠心肌微血管内皮细胞的体外培养

Culture of mouse cardiac microvascular endothelial cells in vitro

目的建立小鼠心肌微血管内皮细胞培养体系。方法 4～6周的清洁级C57小鼠的心室肌,利用胰蛋白酶及Ⅱ型胶原酶消化过滤收集的滤液进行重新悬浮种植于明胶包被的培养瓶中,通过倒置电镜观察细胞的生长形态及生长状态,得出生长曲线,并利用免疫荧光鉴定(心肌微血管内皮细胞特异性抗原vWF)培养出的小鼠心肌微血管内皮细胞。结果通过形态学观察及免疫荧光鉴定证实为小鼠心肌微血管内皮细胞。培养的小鼠心肌微血管内皮细胞第1、2天生长相对缓慢,而到第3、4天细胞呈对数生长,第6、7天细胞达到融合。结论采用明胶包被培养瓶,通过机械剪切、蛋白酶消化、过滤方法,并进行了相关鉴定,可获得较纯的小鼠心肌微血管内皮细胞,这为研究心肌微血管内皮细胞的迁移、血管再生等提供了实验来源。

Objective To establish the cuhure system of mouse cardiac microvascular endothelial cells. Method： The ventricular muscle from 4 to 6 weeks of C57 mice was digested by trypsin and type H collagen,then filtered and c lected the filtrate to grow in culture bottle that was used gelatin to envelop. The morphology and growth curve of mo cardiac microvascular endothelial cells was investigated by using electron microscopy. The immunofluorescence was u to identify culture of mouse primary cardiac microvascular endothelial cells by the expression of ：factor vWF-related al gen. Results The mouse cardiac microvascular endothelial cells were demonstrated by morphological observation and i munofluorescence. Culture of mouse cardiac microvascular endothelial cells grow were relatively slowly in 1 to 2 day grow ogarithmicly in 3 to 4 day, grow cell fusion in 6 to 7 day. Conclusion In the experiment, used gelatin to envelq culture bottle,We can obtain mouse cardiac microvascular endothelial cells with higher purity by mechanical shearil protease digestion, fihering, and relevant identification,thus to provides the source of the study of cardiac microvascL endothelial cell migration and revascularization.