大鼠血管平滑肌细胞的培养和鉴定

Isolation,Culture,and Identification of Rat's Vascular Smooth Muscle Cells

目的:建立一种方便快捷的培养大量大鼠血管平滑肌细胞的方法,为心血管疾病的体外实验研究提供实验材料。方法:采用改良的组织块贴壁法进行原代培养,差异贴壁法纯化细胞,常规胰酶消化法传代,使用倒置相差显微镜进行形态学观察,通过检测平滑肌肌动蛋白对细胞进行免疫组化鉴定。结果:培养4代的平滑肌细胞纯度达96%以上。镜下可见培养细胞呈典型的"峰-谷"样生长,免疫组化染色显示胞质内平滑肌肌动蛋白阳性表达。结论:本研究所介绍的组织贴块培养法较为成熟,可为心血管疾病病因、机制的研究和防治提供一个理想的实验材料。

Objective： To establish a convenient method to provide rat vascular smooth muscle cells （VSMCs） from rat thoracic artery for in vitro study of cardiovascular disease. Methods： The pri- mary culture was obtained by modified tissue-piece inoculation. The passage cells were digested with 0.9,5% trypsin. Morphological changes was observed using an inverted phase contrast mi- croscope and identified by immunohistochemical methods with stained smooth muscle-α-actin. Results： The purity of VSMCs cultured for 4 generations was more than 96%. The passaged VSMCs showed the typically peak-and-valley growth pattern. Immunohistochemical staining showed that mooth muscle actin was expressed in cytoplasm. Conclusion： The method presented here is very simple, and can provide the ideal experimental materials for etiology and prevention study on cardiovascular disease.