Second-passage Japanese quail embryo cell cultures, normal or quantitatively transformed by Rous sarcoma virus, were investigated for phospholipid composition and metabolism. Cells cultivated at low and high population density as well as in the presence or absence of serum, were compared by chemical analysis and in pulse-chase experiments. No differences in the lipid compositions between the normal and the tumor cells or between cells under different culture conditions were detected. In no case was the metabolism of phosphatidylserine or sphingomyelin affected by culture conditions. The metabolism of the choline and ethanolamine glycerophospholipids, however, differed according to culture conditions, whether cells were normal or transformed. Significantly, in normal cells, the breakdown of [32P]phosphate-labeled phosphatidylinositol was slowed when cell growth was restricted, i.e., at high population density or in medium without serum. This effect was not observed in tumor cells under such culture conditions, and cells were not growth inhibited. Release of [32P]phosphate from phosphatidylinositol is the only parameter in the metabolism of phospholipids observed to correlate with growth.