改良组织块贴壁法培养小鼠主动脉血管平滑肌细胞及生物学鉴定?

目的：探讨小鼠主动脉血管平滑肌细胞(VSMC)的原代培养方法及生物学特性,为血管性疾病细胞及分子水平的科学研究提供实验材料。方法分离小鼠胸、腹主动脉,采用改良组织块贴壁法获得主动脉 VSMC,胰蛋白酶消化传代,差速贴壁法进行细胞纯化,倒置相差显微镜观察细胞形态及生长情况,并用苏木素-伊红(HE)染色法和免疫荧光法进行鉴定。结果该方法成功分离出小鼠主动脉 VSMC,细胞生长旺盛,活性良好,呈放射状、典型“峰-谷”样生长,HE 染色细胞呈梭形,细胞质丰富,核大而圆形或椭圆形,细胞免疫荧光显示特异性的细胞质内α-平滑肌肌动蛋白阳性表达。结论本方法简单、经济、可靠,可在体外条件下分离,培养出纯度高、活性良好的主动脉 VSMC。

Improved tissue piece inoculation and identification of mouse vascular smooth muscle cell

Objective To explore the method of primary culture and biological characteristics of aortic vascular smooth mus-cle cells (VSMC)in mice,providing experimental material for cellular and molecular scientific research of vascular disease. Methods Thoracic and abdominal aortas in mice were isolated and VSMC were obtained by using improved method of tissue piece inoculation.Digested with trypsin and passaged,VSMC were purified with differential adherence method.The conditions of cellular morphology and growth were observed under inverted phase contrast microscope,and VSMC were identified with hematoxylin-eosin (HE)staining and immunofluorescence.Results VSMC were isolated successfully and grown vigorously with good bioactivity,the cells had a radial or typical"peak-valley"like growth,and showed fusiform,abundant cytoplasm with large and round or oval nucle-us by HE staining,the expressions of specific cytoplasmicα-smooth muscle actin were positive by immunofluorescence stain.Conclu-sion It can isolate and cultivate VSMC with high purity and good activity under in vitro conditions with simple,economical,reliable method.