We have investigated the content and composition of neutral glycosphingolipids (GSLs) in the cells isolated by enzyme digestion from elastic-hyperplastic and musculo-elastic intimal layers of grossly normal and atherosclerotic regions of human aorta. We have detected three types of neutral GSLs in the intimal cells identified as glucosylceramide, trihexosylceramide and tetrahexosylceramide. We failed to detect lactosylceramide in the intimal cells. The cells of the elastic-hyperplastic layer of grossly normal regions contained trihexosylceramide and tetrahexosylceramide, while glucosylceramide was not detected. Considerable amounts of glucosylceramide were found, and the trihexosylceramide and tetrahexosylceramide content was increased in the cells isolated from atherosclerotic regions. The cells of the musculo-elastic layer of grossly normal intimal regions contained glucosylceramide, trihexosylceramide and tetrahexosylceramide. Cells of the musculo-elastic layer of the fatty streak contained noticeable higher amounts of glucosylceramide, as well as greater amounts of trihexosylceramide and tetrahexosylceramide. Cells of the musculo-elastic layer of the plaque also appeared to contain more glucosylceramide, tetrahexosylceramide, but less trihexosylceramide as compared with grossly normal regions. In both cases cells of the fatty streak exhibited the highest total amount of neutral GSLs, but at the same time the neutral GSL composition of the fatty streak was not similar to GSL composition which is known for human blood monocytes. These findings indicate that elevation of neutral GSL level is observed in cells from atherosclerotic lesions of human aortic intima.