SD大鼠胸主动脉血管平滑肌细胞组织贴块法培养及鉴定

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,4⁶h后加入含20%胎牛血清的DMEM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,56h后加入含20%胎牛血清的DMEM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,5⁷d细胞迁出,107d细胞迁出,10¹4d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。

Adherent culture and identification of smooth muscle cells isolated from SD rat thoracic aorta

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 under sterile conditions.The VSMC were isolated from the SD rat thoracic aorta,primarily cultured4-6hlater with DMEM containing 20% FCS for two weeks,and identified with immunoflurescence staining.Results The VSMC grew out on days 5-7,became nearly 80%confluent on days10-14 after adherent culture,and could thus be passaged.The in vitro cultured VSMC could be passaged to the 20 th generation and grew in a typical"peak valley"mode.Immunoflurescence staining showed that their purity was 95%.Conclusion Adherent culture is a stable,simple and convenient procedure for VSMC isolated from SD rat aorta with a high purity.Immunoflurescence staining can effectively identify VSMC.