Lagarde M, Sicard B, Guichardant M, Felisi O, Dechavanne M
In Vitro. 1984 Jan;20(1):33-7. doi: 10.1007/BF02633329.
Endothelial cells from human umbilical veins were isolated by collagenase treatment. Cells were cultured in the presence of either 20% fetal bovine serum (FBS) or 20% human serum (HS). At confluency, endothelial cell lipids were labeled with tracer concentrations of tritiated arachidonic acid, then extracted and separated into lipid subclasses by thin layer chromatography. The fatty acid composition of each lipid class was determined by glass capillary gas-liquid chromatography analysis and compared to that of cells freshly isolated from the cord (NC cells). The fatty acid compositions differed only in phospholipids. Polyunsaturated fatty acids (PFAs), arachidonic, and linoleic acids were depleted in FBS cell phospholipids and replaced by both stearic and oleic acids. No significant difference could be observed between NC cell and HS cell phospholipids. We conclude that PFAs might be decreased in FBS cells because of the relative paucity of PFAs in FBS as compared to HS. It seems therefore more convenient to cultivate endothelial cells in the presence of HS, especially in respect to their phospholipid content of arachidonic acid, which is the physiological reservoir for prostacyclin synthesis.
通过胶原酶处理分离出人脐静脉内皮细胞。细胞在含有20%胎牛血清(FBS)或20%人血清(HS)的条件下培养。汇合时,用示踪浓度的氚标记花生四烯酸标记内皮细胞脂质,然后提取并用薄层色谱法分离成脂质亚类。通过玻璃毛细管气液色谱分析确定每个脂质类别的脂肪酸组成,并与从脐带新鲜分离的细胞(NC细胞)的脂肪酸组成进行比较。脂肪酸组成仅在磷脂方面有所不同。多不饱和脂肪酸(PFAs)、花生四烯酸和亚油酸在FBS细胞磷脂中减少,并被硬脂酸和油酸取代。NC细胞和HS细胞磷脂之间未观察到显著差异。我们得出结论,由于与HS相比FBS中PFAs相对缺乏,FBS细胞中的PFAs可能会减少。因此,在HS存在下培养内皮细胞似乎更方便,特别是就其花生四烯酸的磷脂含量而言,花生四烯酸是前列环素合成的生理储存库。