Galella G, Marangoni F, Risé P, Colombo C, Galli G, Galli C
Institute of Pharmacological Sciences, University of Milan, Italy.
Biochim Biophys Acta. 1993 Sep 8;1169(3):280-90. doi: 10.1016/0005-2760(93)90252-5.
The human monocytic leukemia cell line THP-1 is depleted of the long-chain n-6, AA, when compared to human monocytes. This reflects the low availability of this FA in the growth medium generally used for cultured cells. The effects of AA, as well as EPA, supplementation of THP-1 cells on the incorporation of these FA in cell PL, especially in PC and PE, was investigated. In addition the incorporation of labeled AA in PL from THP-1 cells was compared to that in human monocytes. Measurements were done through HPLC separation of PL, detected by UV absorption and radioactivity, FA analysis by GC and characterization of PC subclasses by FAB-MS. Marked differences were observed in the incorporation of the two FA in cell PL, particularly two PC subclasses, and in the accumulation in individual PL after supplementation of THP-1 cells. Accumulation of AA and EPA in THP-1 cells appeared to be mutually independent. The incorporation of AA was also quite different in THP-1 from that in monocytes. Thus, characterization of the FA content in lipids of cultured cells is an essential requirement for optimal utilization of these cells.
与人类单核细胞相比,人单核细胞白血病细胞系THP-1缺乏长链n-6脂肪酸花生四烯酸(AA)。这反映了在通常用于培养细胞的生长培养基中这种脂肪酸的可利用性较低。研究了补充AA以及二十碳五烯酸(EPA)对THP-1细胞中这些脂肪酸掺入细胞磷脂(尤其是磷脂酰胆碱(PC)和磷脂酰乙醇胺(PE))的影响。此外,还比较了THP-1细胞中标记的AA掺入磷脂的情况与人单核细胞的差异。通过高效液相色谱(HPLC)分离磷脂,紫外吸收和放射性检测,气相色谱(GC)分析脂肪酸以及快原子轰击质谱(FAB-MS)表征PC亚类进行测量。在细胞磷脂中,尤其是两个PC亚类中,两种脂肪酸的掺入以及THP-1细胞补充后单个磷脂中的积累存在明显差异。AA和EPA在THP-1细胞中的积累似乎相互独立。THP-1细胞中AA的掺入也与单核细胞中的掺入有很大不同。因此,表征培养细胞脂质中的脂肪酸含量是最佳利用这些细胞的基本要求。