目的探讨一种方便、快捷、大量原代培养大鼠血管壁中膜平滑肌细胞的方法,为心血管疾病的体外研究提供可靠的实验材料。方法利用改良的组织贴块法进行血管壁中膜平滑肌细胞原代培养,使用倒置相差显微镜观察细胞形态,并用免疫荧光染色鉴定分离培养的细胞。结果改良的组织贴块法成功培养出大鼠血管壁中膜平滑肌细胞,3 d~5 d可见细胞爬出,7 d~9 d即可传代。镜下见细胞以长梭形为主,呈典型"峰-谷"样生长,免疫荧光染色鉴定阳性细胞率在95%以上。结论本研究介绍的改良组织贴块法技术成熟,具有操作简单、方便等优点,使用该法不仅可以获得纯度高、活性好的血管中膜平滑肌细胞,而且缩短了培养周期。
Objective To investigate the primary culture and identification of the smooth muscle cells (SMC) from the medial of vascu- lar wall in rats. Methods SMC from the medial of vascular wall were cultured by modified tissue culture. The cellular morphology was observed under inverted phase contrast microscope. SMC were isolated and identified with immunofluorescence. Results SMC were isolated successfully and cultured. The cells had a typical peak - valley like growth, and showed fusiform. The positive rate of cells i- dentified with immunofluorescence were more than 95%. Conclusion The modified tissue culture can isolate and cultivate SMC with high purity and good activity under in vitro conditions with simple, reliable method.