SD 大鼠胸主动脉血管平滑肌细胞组织贴块法培养及鉴定

摘要

Objective To identify the vascular smooth muscle cells (VSMC) isolated from SD rat thoracic aorta after adherent culture .Methods The thoracic aorta was isolated from SD rats un-der sterile conditions .The VSMC were isolated from the SD rat thoracic aorta ,primarily cultured 4-6 h later with DMEM containing 20% FCS for two weeks ,and identified with immunoflures-cence staining .Results The VSMC grew out on days 5-7 ,became nearly 80% confluent on days 10-14 after adherent culture ,and could thus be passaged .The in vitro cultured VSMC could be passaged to the 20th generation and grew in a typical“peak valley”mode .Immunoflurescence stai-ning showed that their purity was 95% .Conclusion Adherent culture is a stable ,simple and con-venient procedure for VSMC isolated from SD rat aorta with a high purity .Immunoflurescence staining can effectively identify VSMC .%目的：建立SD大鼠胸主动脉血管平滑肌细胞（vascular smooth muscle cells ，VSMC）组织贴块培养法并进行鉴定。方法无菌条件下取大鼠胸主动脉，4～6 h后加入含20％胎牛血清的DM EM培养液，原代培养2周并用免疫荧光染色对培养的VSMC进行鉴定。结果组织贴块法成功培养了大鼠胸主动脉VSMC ，5～7 d细胞迁出，10～14 d细胞融合至80％进行传代。体外培养的VSMC可传至20代，培养的细胞呈典型的“峰谷”样生长，免疫荧光染色鉴定细胞纯度达95％。结论组织贴块法培养大鼠胸主动脉VSMC培养体系稳定，操作简单、方便，纯度较高，免疫荧光是鉴定VSMC的较佳方法。