酶消化法分离培养家兔血管平滑肌细胞

目的探讨胶原酶分离并培养家兔血管平滑肌细胞的方法。方法取家兔腹主动脉，0．2％胶原酶Ⅰ消化分离血管平滑肌细胞并进行培养，相差显微镜观察培养细胞的形态，免疫组化染色法检测细胞胞浆内α-肌动蛋白（α-actin）的表达。透射电镜观察血管平滑肌超微结构。结果相差显微镜下细胞呈长梭形，未见明显异型细胞，α-actin胞浆染色阳性细胞数占总细胞数的99％以上。电镜显示VSMC内肌丝丰富，线粒体、高尔基复合体等细胞器较少，VSMC呈现分化的收缩表型。结论胶原酶可消化分离培养血管平滑肌细胞，且有成活细胞数多、传代周期短的特点，可用于血管平滑肌表型研究。

ISOLATION AND CULTURE VASCULAR SMOOTH MUSCLE CELLS OF RABBIT BY ENZYMATIC DIGESTION METHOD

Objective To investigate the method of isolation and culture rabbit＇ s vascular smooth muscle cells （VSMC） by collagenase. Methods 0.2% collagenase Ⅰ was used to digest and isolate the abdominal aortic smooth muscle cells of rabbit and then cultured. Contrast microscope was used to observe in morphology of cultured cells, α -actin expression in cytosol measured by immunohistochemical staining. Ultrastructure of VSMC was ob- served by transmission electron microscope. Results The cells shown long spindle - shape and the dysplasia cells were not detected and cells that observed α - actin stained were more than 99% . Electron microscope shown that the myofilament in cytosol of VSMC were abundant and organelle such as endoplasmic reticulum, Golgi＇ s complex, mitochondrion were less and VSMC has contractile phenotype. Conclusion Collagenase can digest and isolate VSM and also has advantages that make cells easy to live and short period for passaging. This method can be used for study phenotype of VSMC.