Cholesterol content and sterol synthesis in human skin fibroblasts and rat aortic smooth muscle cells exposed to lipoprotein-depleted serum and high density apolipoprotein/phospholipid mixtures

Abstract

Confluent cultures of human skin fibroblasts and rat aortic smooth muscle cells were shown to lose 15-27% of their cellular cholesterol upon replacement of the fetal calf serum with human high density lipoprotein (50 mug cholesterol/ml) or lipoprotein-depleted serum at a concentration equivalent to 40% whole serum. Addition to the latter medium of high density apoliproprotein/phospholipid mixtures resulted in further enhancement of cellular cholesterol loss which was evident by 12 h of incubation. Human skin fibroblasts that had been enriched in cholesterol by previous incubation with low density lipoprotein lost their cholesterol in the presence of a high density apolipoprotein/sphingomyelin mixture as readily as non-enriched cells. Concomitant with the marked cholesterol depletion there was a stimulation of sterol synthesis from acetate. The more pronounced loss of cellular cholesterol induced by the presence of phosphatidylcholine or sphingomyelin resulted in a greater incorporation of acetate into sterol in both smooth muscle cells and skin fibroblasts. The present findings indicate that peripheral cells, in spite of their capacity to synthesize cholesterol, depend on exogenous cholesterol for the maintenance of normal levels. It is suggested that the native cholesterol "acceptor" in the lipoprotein-depleted serum is an apolipoprotein which under the experimental conditions can form a complex with phospholipids and might also represent the physiological cholesterol "acceptor" in peripheral lymph.