人脐血内皮祖细胞体外培养扩增的初步研究

Culture and proliferation of human umbilical cord blood endothelial progenitor cells in vitro

目的探索人脐血内皮祖细胞(EPCs)代替角膜内皮细胞作为种子细胞的可行性。方法利用密度梯度离心法从人骨髓中分离单核细胞,接种至纤维连接蛋白铺层的培养皿中,EGM-2培养液培养。检测人骨髓EPCs摄取乙酰化低密度脂蛋白(AcLDL)及结合荆豆凝集素1(UEA1)的能力;免疫荧光法检测CD34、CD133和vWF的表达情况。结果人脐血EPCs呈短梭形或多边形,纤维连接蛋白铺层可提高EPCs的贴附率;传代不影响细胞的形态和增殖,可大量扩增。人脐血EPCs能摄取AcLDL,结合UEA1,表达CD34、CD133和vWF。结论人脐血EPCs有望成为较理想的代替角膜内皮细胞的种子细胞。

Objective To investigate the feasibility of human umbilical cord blood endothelial progenitor cells （EPCs） in substitute for corneal endothelial cells as seed cells. Methods Mononuclear cells were isolated from human umbilical cord blood by density gradient centrifugation, and were plated in endothelial growth medium on fibronectin coated culture dish with EGM-2 as culture fluid. The ability of EPCs in uptaking acetylated low density lipoprotein （AcLDL） and binding Ulex Europaeus Lectin 1 （UEA1） was determined, and the expression of CD133, CD34 and vWF was detected by immunofluorescence method. Results The human umbilical cord blood EPCs exhibited polygonal or short spindle shape. Fibronectin-coated dish facilitated the adhesion of EPCs. Cellular morphology and proliferation ability were not changed by passage. The human umbilical cord blood EPCs could uptake AcLDL, bind UEA1, and express CD133, CD34 and vWF. Conclusion Human umbilical cord blood EPCs may be favorable candidate in replacement of corneal endothelial cells as seed cells.