改良组织块贴壁法培养大鼠主动脉平滑肌细胞及鉴定

目的以简捷、高效的方法获取原代和传代的大鼠血管平滑肌细胞(VSMC),为进一步的科学研究提供实验材料。方法采用改良的组织块贴壁法培养原代大鼠VSMC,2.5 g.L-1胰蛋白酶消化传代,并用相差显微镜、电镜、荧光显微镜及激光共聚焦显微镜对细胞进行鉴定。结果95%的组织块接种成活,相差显微镜下细胞呈典型的"峰-谷"状生长,胞体长梭形;电镜下可见粗、细肌丝沿细胞长轴平行排列;荧光显微镜及激光共聚焦显微镜检测α-肌动蛋白可见细胞胞浆内呈阳性显色,细胞核不显色。结论本方法培养VSMC简便、易操作,获得的细胞纯度和成活率较高。

Culture and identification of rat's vascular smooth muscle cells with modified tissue-piece inoculation

Objective To obtain vascular smooth muscle cells of original generation and passage of rat in simple and effective way for the related researches.Methods The original generation cells were obtained by modified tissue-piece inoculation and with 0.25% trypsin digestion to obtain passage cells.The cells were identified by contrast phase microscope,electron microscope,fluorescence microscope and confocal microscopy.Results 95% inoculated tissue pieces were survived.Under contrast phase microscope,cells showed the typical "peak-valley" morphological,cell bodies were fusiform shape.Under electron microscope thick myofilament and thin filament ranged parallelly along cell macroaxis.Under fluorescence microscope and confocal microscopy,endochylema of α-actin was been seen that it was stained positive.And the nucelus wasn′t been stained.Conclusion It is a simple and easy way to obtain highly purified VSMC and the survival rate is high.