人血管内皮细胞在体外连续增殖传代的研究

Studies of Serials Propagation of Human Endothelial Cells in Vitro

作者用胶原酶消化法自人脐带静脉分离获取内皮细胞,由牛下丘脑提取内皮细胞生长因子(ECGF),以改进的方法对内皮细胞进行培养,建立起在体外连续增殖传代的人血管内皮细胞(HEC)的培养模型。并就ECGF和人纤维粘连蛋白(HFN)对HEC生长的生物学效应进行观察和比较。结果表明,①HEC在低密度(5×10~3/cm^2)情况下,即对ECGF(100～300mg/L)有良好的增殖反应,可在体外连续增殖传至10代;②HFN对HEC增殖无直接促进作用,但预先用其涂铺培养瓶底面,有利于细胞贴壁:③ECGF和HFN不能替代小牛血清。此培养体系的建立,将为内皮细胞的生理和病理研究提供便利条件。

Successful serial propagation of human endothelial cells (HFCs) in vitro was established in the present investigation. HECs were isolated from human umbilical vein by collagenase digestion and identified as endothelial cells by morphologic and immunologic criteria. The cells were grown in medium RPMI-1640 supplemented either with fetal bovine serum(FBS) and endothelial cell grwth facter (FCGF), bovin Hypothalamic extract, in the presence or absence of human fibronectin (HFN) Matrix or with FBS and/or HFN in the presence or absence of ECGF. The results of endothelial cell assays demonstrated that ECGF could stimulate the serial propagation of low-seed-density ( 5 × 10~3 cells/cm^2 ) HEC in vitro, and ECGF is biologically active at 100～300 mg/L, that the role of HFN matrix simply provides a surface for optimal endothelial cell attachment and does not influence the cell growth and that FBS is essential for HEC growth. The establishment of this culture sysyem may attribute to the further study of endothdial cells in physiology and pathology.