大鼠胸主动脉平滑肌细胞的组织贴块法培养及鉴定

目的:建立大鼠胸主动脉平滑肌细胞的组织贴块培养法.方法:取大鼠胸主动脉,切成1 mm×1 mm×1 mm的小块,均匀地种植于培养瓶壁上,约50 min后加入含有20%小牛血清的PRMI-1640培养基进行培养.结果:培养第5天时,可见少量梭形或长梭形细胞自组织块周围游出,至培养3周细胞融合成片,呈"峰、谷"状生长.肌动蛋白免疫组化染色,＞98%的细胞染色阳性,透射电镜观察,于基膜下和胞浆内可见密斑和密体.VG染色也证实几乎所有的细胞均为平滑肌细胞.结论:应用细胞贴块法培养大鼠平滑肌细胞,操作简单,结果稳定,纯度较高,具有应用价值.

Attachment-block culture and identification of smooth muscle cells from thoracic aorta in rats

Objective:To investigate the method of attachment block culture of smooth muscle cells from thoracic aorta in rats.Methods:The thoracic aorta of rats were separated carefully and cut into small pieces about 1 mm 3,and then these pieces were attached on the wall of culture bottle.After 50 minutes,those tissue pieces were cultivated in PRMI 1640 with 20% calf serum.Results:A few of cells had grown on the bottem of culture bottle 5 days late,and had developed into groups like peak valley in shape after cultivated for 3 weeks. All cells almost were smooth muscle cells identified through VG stain and Anti α actin immunohistochemistry stain.Dense dots and dense bodies were observed beneath the cell matrix under the electron microscope.Conclusion:Attachment block culture of rat smooth muscle cells is simple in operation,reliable in result,high in purity and useful as a cellular model for study and prevention of vascular diseases.