Melanosomes synthesized by embryonic chick retinal pigment cells in vitro are of a different size and shape than those made in ovo and in vivo. Ultrastructurally, the melanosomes made by cultured cells are small, spherical, contain an irregular internal matrix, and appear similar to the pheomelanosomes found in red hair and feathers. This modulation in melanosome ultrastructure in cultured retinal pigment cells does not appear to be caused by an irreversible change in differentiated capability but to a response of cells to the culture environment. Cells grown in embryo extract-free and serum-free defined media were found to synthesize melanosomes of normal size and shape in vitro, although at a much reduced number. The predominant melanosome type found was still the smaller, spherical variety. It appears that some still unknown factors in the culture environment have produced a shift to a different type of melanogenesis.