The cells that surround and support the seminiferous epithelium of the tubules in the testis, i.e., the peritubular or myoid cells, are known to contain relatively high amounts of cellular retinol-binding protein (CRBP). This suggests that they may play an important role in the movement or metabolism of retinol (vitamin A alcohol), which is required for the maintenance of spermatogenesis. Peritubular cells in culture, isolated from the testes of the 20-day-old rat, maintained high levels of CRBP and had the ability to internalize retinol from retinol-binding protein (RBP), the blood transport protein for retinol, in a manner suggesting a receptor-mediated process. Little of the internalized retinol was esterified, in contrast to what occurs in other cell types that contain high amounts of CRBPs, and very little, if any, lecithin-retinol acyltransferase activity was present in microsomes obtained from the cultured cells. The cells did, however, have the ability to synthesize and release their own RBP to the medium. This suggests that retinol from the blood may actually reach the seminiferous epithelium by passing across the peritubular cell, released on a new molecule of RBP, rather than by entering into the tubule bound to the preexisting RBP present in the interstitial fluid.