Insulin has potent effects on osteoblast function both in vivo and in vitro. In various insulin‐sensitive tissues, stimulation of glucose transport and metabolism are hallmarks of insulin action, and have been postulated to play a role in insulin regulation of cellular function. However, insulin effects on glucose metabolism in osteoblast‐like cells have not been demonstrated. Therefore we examined the in vitro effects of insulin on hexose uptake in an osteoblast‐enriched rat bone explant preparation. Uniform 5‐mm‐diameter punch sections were obtained from the cartilage‐free frontal portions of the calvaria of 3‐day‐old rats, and the periosteum was removed. The resulting sections contained a highly enriched population of osteoblast‐like cells as determined by histologic criteria, elimination of calcitonin‐stimulatable cAMP generation, and enhancement of PTH‐stimulatable cAMP generation per microgram of DNA. Sections were incubated for 24 hr at 37°C in BGJb medium and then transferred to modified glucose‐free Krebs‐Ringer bicarbonate buffer for 2‐deoxy‐D‐glucose (2‐DG) uptake studies. ³H‐2‐DG uptake was linear with time over 60 min, temperature sensitive, and inhibited by 5 mA/phloridzin. Kinetic analysis of 2‐DG uptake at 25°C demonstrated a saturable transport mechanism with a K_m of 2.2 mM, similar to that observed for 2‐DG transport in other tissues. Studies of competitive inhibition by other sugars demonstrated a transport specificity for 2‐DG that was comparable to that previously observed in fat and muscle cells. 2‐DG uptake was rapidly stimulated by insulin (0.5 nM‐1 μM), with maximal insulin effect observed at 50 nM. Concanavalin A (10–50 μg/ml) produced a similar stimulation of 2‐DG transport. Both basal and insulin‐stimulated uptake were inhibited reversibly by cytochalasin B (50 μM). Insulin stimulation of 2‐DG uptake was the result of increased transport V_max, with K_m remaining constant. These studies provide the first demonstration that the cell population of osteoblast‐enriched bone explants possesses an insulin‐stimulatable glucose transport system with characteristics similar to those found in other insuln‐sensitive tissues.