自发性高血压大鼠血管平滑肌细胞的培养及鉴定

摘要

Objective :Culture and identification of Spontaneously hypertension rat aortic smooth muscle cells ,to ana-lyze cell growth and biological characteristics for the research of the proliferation of the vascular diseases .Methods :The culture of was done by tissue-piece inoculation .The cultured cells were observed and identified by morphology and im-munohistochemistry with phase contrast microscope and immunohistochemical staining respectively .The cell viability was determined with Typan blue .The survival ratio ,growth and proliferation characteristics of passage VSMC were measured by MTT .Results :Cells growth curve were similar to“S”shape ,passage cycle was 7~9 days ,the growth char-acteristics of VSMC assumed the model“peak and valley”.MTT assay showed that optical density change cell growth was obvious at 7~9 days .SM-action demonstrated these cell were positive .Positive results than 4 generation 2 .Con-clusion:The method of tissue piece in oculation culture of VSMC is simple economic and efficient .It provides an ideal cell model for the research of the proliferation of vascular diseases .Along with the wedding process cell biological char-acteristics change .%目的：培养、鉴定自发性高血压大鼠主动脉平滑肌细胞，了解生长特性，为血管增生性疾病的治疗研究提供试验材料。方法：采用组织贴块法培养大鼠胸主动脉血管平滑肌细胞，用倒置相差显微镜观察其生长情况，用免疫组化染色做鉴定，台盼蓝染色进行活力检测，绘制生长曲线、MTT法测定VSMC细胞的成活率、生长、增殖特征。结果：细胞生长曲线近似“S”形，VSMC传代周期为7～10d ，呈典型“峰-谷”状生长，MTT法显示细胞生长第7～9天光密度值变化较明显，免疫组化染色显示胞浆内平滑肌肌动蛋白阳性表达，第2代阳性结果强于4代。结论：正确的利用组织块可以简单、经济、高效地培养出VSCM ，为VSCM的治疗提供了理想的细胞模型。随传代过程细胞生物学特性发生改变。