As a first step in studying the effects of membrane lipid modification on complex cellular functions we have modified the membrane fatty acid composition of the neuroblastoma X glioma hybrid clone, NG108-15. These cultured cells were chosen because they exhibit many complex neuronal functions in vitro. Unsaturated fatty acids (oleate, linoleate, linolenate and arachidonate) were accumulated, metabolized and esterified by the cells. These unsaturated fatty acids stimulated cell growth, whereas saturated fatty acids were toxic to the cells. Changes as large as 40-fold in the ratio of monounsaturated/polyunsaturated fatty acids in the membrane phospholipids were produced by addition of fatty acids directly to serum-containing culture medium. As a result of the exposure of NG108-15 cells to unsaturated fatty acids the amount of phosphatidylethanolamine in the cells was increased by as much as 60%. Polyunsaturated fatty acids also caused a small decrease in the membrane cholesterol/phospholipid molar ratio. These experiments demonstrate that large changes in membrane fatty acid composition can be created in clonal cells capable of differentiated neuronal activities. Additional changes in membrane lipid composition also appear to be induced by these manipulations. The question of the importance of specific membrane lipid composition to neuronal cellular function now can be addressed.