内皮祖细胞培养条件的正交设计研究

Study on culture condition of endothelial progenitor cells by orthogonal design

目的:研究内皮祖细胞(EPCs)培养的适宜条件。方法:以L827正交表安排影响FPCs扩增贴壁的几个主要因素,光镜下观察集落形成,流式细胞术分析CD34、VE-Cadherin双阳性细胞,以此为指标分析各因素对EPCs培养的影响。结果:血管内皮生长因子(VEGF)及接种密度对集落形成影响显著, 内皮细胞生长添加剂(ECGS)及纤维连接蛋白皿底包被无影响。4个因素对EPCs得率均有影响,且VEGF 及接种密度之间尚有交互作用。结论:EPCs培养的适宜条件为:皿底包被纤维连接蛋白,5×106/cm2接种密度,VEGF 10 ng/ml及ECGS 150μg/m1。

Objective： To investigate the proper culture condition of endothelial progenitor cells （EPCs）. Methods： L82^7 orthogonal design list was used to arrange the four main factors which affect the proliferation and adhesion of cultured EPCs. Cluster forming was observed under light microscope. The expression of CD34 and VE-Cadherin was performed by FACS.The influence of each factor then was analyzed. Results： Factor VEGF and cell density had significant influence on cluster forming, while factor ECGS and fibronectin did not affect it. Four factors all had significant influence on number of EPCs, and there also was an interaction between factor VEGF and fibronectin. Conclution： The proper culture condition of endothelial progenitor cells is： coating culture dish with fibronectin, 5 × 10^6/cm^2 cell density, VEGF 10ng/ml and ECGS 150μg/ml.