Distraction osteogenesis (DO) is a surgical procedure in which applied strain stimulates new bone growth; however, the underlying mechanisms by which bone cells respond to load are still uncertain. An organ culture model of DO was developed and validated by using linear distraction on the femoral shafts of 5 day old Wistar rats. Two loading regimes were utilized: distracting the bones for 2 hrs on day 1 (GRP I); distracting the bones for 2 hrs on days 1, 3, and 5 (GRP II). After 1 week in culture, the bones were compared to unloaded contralateral controls and assessed for changes. Structural, dimensional, massing, micro-CT, areal, and viability properties were obtained from testing. Relative to paired controls, distracted bones demonstrated an increase in failure load (9.15% GRP I, 18.85% GRP II), increase in stiffness (31.28% GRP I, 53.21% GRP II), increases in areal and polar moments of inertia, and viability (6.21% GRP I, 13.02% GRP II). Our results suggest that DO can be modeled successfully with an organ culture, and continued use of this system will help to gain insight into the mechanisms and pathways by which distraction osteogenesis occurs.