大鼠主动脉内皮细胞的分离培养和鉴定

Isolation Culture and Identification of Rat Aortic Endothelial Cells

目的:探索大鼠主动脉原代内皮细胞体外培养方法,为体外研究提供细胞模型。方法:分离大鼠主动脉,直接贴壁于培养皿中,荧光倒置显微镜观察细胞形态,免疫组化Ⅷ因子相关抗原染色鉴定细胞。结果:约24小时组织块边缘有游离的新生细胞长出,7天即融合成片。消化传代后细胞呈短梭形或三角形,单层生长,铺路石状,Ⅷ因子表达阳性,呈指数增殖。冻存后复苏细胞活性均超过90%。结论:用贴壁法成功建立了大鼠血管内皮细胞体外培养方法,冻存细胞存活率高,为体外研究提供了稳定的模型。

Objective： To investigate the culture methods of primary rat aortic endothelial cells in vitro. Methods： Rat aorta was isolated, directly attached to the wall in a petri dish, and inverted microscope cell morphology. Immunohistochemistry factor VllI-related antigen was used to stain cells. Results： Organization block edge free new cells grow, after approximately 24, and integrated into the film 7d later. Cells were subcultured short spindle-shaped or triangular, monolayer, cobblestone, positive expression of factor VIII, the exponential proliferation. Recovery was more than 90% cell viability after cryopreservation. Conclusion： Vascular endothelial cells in vitro method was successfully established by adherent, and the survival rate of frozen cells was high, which providing a stable model for in vitro studies.