Kaji K
Department of Integrated Research, Tokyo Metropolitan Institute of Gerontology, Japan.
Hum Cell. 1988 Jun;1(2):188-97.
The purpose of this review is to introduce a simple and inexpensive culture method for human umbilical blood vessel endothelial cells. The medium used is MCDB-104 supplemented with 10% fetal bovine serum, 70 ng/ml endothelial cell growth factor from new-born bovine brains, 10 ng/ml murine epidermal growth factor, and 100 micrograms/ml heparin. The culture dishes are coated with gelatin. Under these conditions, endothelial cells from human vessels were grown with doubling times of 18-22 hrs and reached saturation densities of 8-12 x 10(4) cells/cm2. To determine the lifespan of the endothelial cells, the cells were serially subcultivated weekly at an inoculum size of 1,000 cells/cm2. Human endothelial cells from umbilical vein and artery were grown for 21 to 37 passages with 55 to 125 population doublings. This culture method seems to be useful for studying cell proliferation and functions of human endothelial cells.
本综述的目的是介绍一种简单且成本低廉的人脐血管内皮细胞培养方法。所用培养基为添加了10%胎牛血清、70 ng/ml新生牛脑来源的内皮细胞生长因子、10 ng/ml小鼠表皮生长因子以及100 μg/ml肝素的MCDB - 104。培养皿用明胶包被。在这些条件下,人血管内皮细胞以18 - 22小时的倍增时间生长,达到8 - 12×10⁴个细胞/cm²的饱和密度。为确定内皮细胞的寿命,细胞每周以1000个细胞/cm²的接种量进行连续传代培养。来自脐静脉和动脉的人内皮细胞传代培养21至37代,群体倍增55至125次。这种培养方法似乎对研究人内皮细胞的细胞增殖和功能很有用。