Björntorp P, Hansson G K, Jonasson L, Pettersson P, Sypniewska G
J Lipid Res. 1983 Feb;24(2):105-12.
Endothelial cells from rat epididymal fat pad capillaries were isolated from rats immediately after weaning. The cells were obtained after an initial brief incubation with collagenase under conditions of minimal breakage of cells. Adipocytes were removed by flotation and endothelial cells were then obtained as cell aggregates by fractional filtration procedures whereby intact tissue as well as free cells were removed. These aggregates were then dispersed and cultured in supplemented medium 199 whereby a monolayer of cells with a growth pattern, numerous pinocytotic vesicles, and intercellular junctions typical of endothelial cells were obtained. Minor contaminations of precursor cells to adipocytes were absent after one subculture. Here greater than 95% of the cells showed the presence of Factor VIII. Further subcultures produced nonhomogenous cells and decreasing rates of replication. The endothelial cells showed a very low rate of triglyceride synthesis and release, and collected no visible lipid upon prolonged cultures in the presence of an abundance of triglyceride substrate. They bound lipoprotein lipase from rat adipocytes, whereby the lipase was stabilized. This binding was released by heparin, and the cells did not synthesize the enzyme.
大鼠附睾脂肪垫毛细血管内皮细胞在断奶后立即从大鼠中分离出来。细胞是在与胶原酶进行初始短暂孵育后获得的，孵育条件是尽量减少细胞破碎。通过浮选去除脂肪细胞，然后通过分级过滤程序以细胞聚集体的形式获得内皮细胞，在此过程中去除完整组织以及游离细胞。然后将这些聚集体分散并在补充的199培养基中培养，从而获得具有典型内皮细胞生长模式、大量胞饮小泡和细胞间连接的单层细胞。一次传代培养后不存在脂肪细胞前体细胞的轻微污染。此处超过95%的细胞显示存在因子VIII。进一步传代培养产生了异质性细胞且复制速率降低。内皮细胞显示出极低的甘油三酯合成和释放速率，并且在存在大量甘油三酯底物的情况下长时间培养后未收集到可见脂质。它们结合来自大鼠脂肪细胞的脂蛋白脂肪酶，从而使该脂肪酶稳定。这种结合可被肝素释放，并且细胞不合成该酶。
