Alpert S E, Walenga R W
Department of Pediatrics, Case Western Reserve University, Rainbow Babies and Children Hospital, Cleveland, Ohio 44106.
Exp Lung Res. 1991 Jan-Feb;17(1):1-15. doi: 10.3109/01902149109063277.
Maintenance of serum-free conditions for the culture of TE or other airway epithelial cells provides a defined environment in which to explore the regulation of cellular functions. Yet TE cells appear to be dependent on the medium for essential, if not all, polyunsaturated fatty acids. At present, some laboratories routinely use serum to support the growth of airway epithelial cells, presumably in part through recognition that cells of mammalian origin require an exogenous source of lipids. While 5% FBS can increase the linoleic and arachidonic acid content of cultured rabbit and human TE cells, it does not fully restore the fatty acid composition of cultured TE cells to that of freshly isolated cells, particularly in the case of human TE cells. Equally good, if not better, repair of membrane fatty acid composition can be achieved by addition of a defined, commercial non-serum source of lipids (Excyte III) plus exogenous arachidonic acid. Cultured TE cells maintained in serum-free medium have been shown to be deficient in prostaglandin and HETE production, both at baseline and in response to physiological stimuli compared to TE cells with greater endogenous content of arachidonic acid. Differences between lipid supplemented and unsupplemented cultured TE cells in cAMP response to PGE2 and in susceptibility to hyperoxic injury have been observed. Other cellular functions regulated by the fatty acid composition of membrane lipids may also be impaired in lipid unsupplemented cells. It is evident that the maintenance of as normal as possible membrane fatty acid content is essential to the use of cultured TE cells as experimental models of airway epithelium.
维持无血清条件培养气管上皮(TE)细胞或其他气道上皮细胞,可为探索细胞功能的调控提供一个明确的环境。然而,TE细胞似乎依赖培养基来获取必需的(即便不是全部)多不饱和脂肪酸。目前,一些实验室常规使用血清来支持气道上皮细胞的生长,推测部分原因是认识到哺乳动物来源的细胞需要外源性脂质。虽然5%胎牛血清(FBS)可增加培养的兔和人TE细胞中亚油酸和花生四烯酸的含量,但它并不能使培养的TE细胞的脂肪酸组成完全恢复到新鲜分离细胞的水平,尤其是人TE细胞。通过添加一种特定的、商业化的非血清脂质来源(Excyte III)外加外源性花生四烯酸,可实现同样好(如果不是更好)的膜脂肪酸组成修复。与花生四烯酸内源性含量更高的TE细胞相比,维持在无血清培养基中的培养TE细胞已被证明在基础状态以及对生理刺激的反应中,前列腺素和羟二十碳四烯酸(HETE)的产生均存在缺陷。在补充脂质和未补充脂质的培养TE细胞之间,已观察到对前列腺素E2(PGE2)的环磷酸腺苷(cAMP)反应以及对高氧损伤的易感性存在差异。膜脂质脂肪酸组成所调节的其他细胞功能在未补充脂质的细胞中也可能受损。很明显,维持尽可能正常的膜脂肪酸含量对于将培养的TE细胞用作气道上皮的实验模型至关重要。