Gorfien S, Spector A, DeLuca D, Weiss S
GIBCO BRL Cell Culture R&D, Grand Island, New York 14072.
Exp Cell Res. 1993 Jun;206(2):291-301. doi: 10.1006/excr.1993.1149.
Endothelial cells grown in vitro are widely used for the study of vascular physiology and pathology. The high levels of serum supplementation normally used in endothelial cell culture may create problems in experimental design or in interpretation of results. A serum-free medium (SFM) has been designed specifically for culture of nonhuman vascular endothelial cells. Growth as well as retention of various physiological markers in this medium was demonstrated in large-vessel endothelial cells from bovine, porcine, ovine, and canine species. Cells from all species tested took up DiI-acetylated LDL. Factor VIII-related antigen was demonstrated in cultures of bovine and canine cells. Bovine and porcine cells were shown to have retained the ability to produce prostacyclin. Further supplementation of the SFM with various lipid-containing additives failed to stimulate either growth or prostacyclin production. PDGF produced by bovine, porcine, and canine endothelial cells was measured and found to be present at lower concentration than in serum supplemented cultures. All cells grown in SFM exhibited the histiotypic "cobblestone" morphology normally associated with cultured endothelial cells. While not optimized for culture of human endothelial cells, short-term growth < or = 4 passages) of human umbilical vein endothelial cells was achieved by supplementing the SFM with hydrocortisone, bovine pituitary extract, and epidermal growth factor.
体外培养的内皮细胞被广泛用于血管生理学和病理学研究。内皮细胞培养中通常使用的高浓度血清补充剂可能会在实验设计或结果解释方面产生问题。一种无血清培养基（SFM）专门设计用于培养非人类血管内皮细胞。在来自牛、猪、羊和犬类的大血管内皮细胞中，已证明该培养基中各种生理标志物的生长及保留情况。所有测试物种的细胞都摄取了DiI-乙酰化低密度脂蛋白。在牛和犬细胞培养物中证实了因子VIII相关抗原。牛和猪细胞显示保留了产生前列环素的能力。用各种含脂质添加剂进一步补充SFM未能刺激生长或前列环素的产生。测量了牛、猪和犬内皮细胞产生的血小板衍生生长因子（PDGF），发现其浓度低于血清补充培养物中的浓度。在SFM中生长的所有细胞均表现出通常与培养的内皮细胞相关的组织型“鹅卵石”形态。虽然未针对人类内皮细胞培养进行优化，但通过向SFM中补充氢化可的松、牛垂体提取物和表皮生长因子，实现了人脐静脉内皮细胞的短期生长（≤4代）。
