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采用密度梯度离心法分离鸡血管内皮祖细胞,通过Dil-acLDL和FITC-UAEI双阳性鉴定血管内皮祖细胞。在三种不同的培养基即培养基A、培养基B、培养基C中进行体外扩增培养,经生长曲线、扩增能力和克隆形成数量三方面评价这三种培养基,结果表明培养基A在体外最适合鸡血管内皮祖细胞的体外扩增。细胞生长曲线呈现明显的“S”形,细胞群体倍增时间约为23.4 h,体外扩增可至9代,细胞克隆形成总数为38。该研究为血管内皮祖细胞体外增值与临床应用奠定了实验依据。
Vascular endothelial progenitor cells were isolated by density gradient centrifugation and endothelial progenitor cells were identified by Dil-acLDL and FITC-UAEI double positive staining. In three different mediums, medium A, medium B and medium C, in vitro expansion and cultivation, the three mediums were evaluated in terms of growth curve, amplification ability and number of colony formation. The results showed that medium A is the most suitable in vitro expansion of chicken vascular endothelial progenitor cells in vitro. Cell growth curve showed obvious “S” shape, cell population doubling time was about 23.4 h, in vitro expansion up to 9 generations, the total number of cell clones formed 38. This study laid the experimental foundation for the value-added and clinical application of endothelial progenitor cells in vitro.