Using 15 mid-term human fetuses, we examined the role of the spine anterior and posterior longitudinal ligaments (ALL, PLL) in ossification of the lumbar vertebral body. By 18 weeks, a pair of calcified tissue or cortical walls had developed on the anterior and posterior sides of the ossification center. These calcified cortical walls were more highly eosinophilic than trabecular or woven bone in the ossification center. Vimentin-positive osteoblasts were arranged in line along the outer surface of the walls. However, few CD68-positive osteoclasts were evident around the walls, suggesting that the calcification in the walls was similar to periosteal ossification. The anterior cortical wall was connected tightly with the ALL by fiber bundles, but the posterior wall was separated from the PLL by the basivertebral (central) vein and loose tissues. Notably, by 30 weeks, the anterior cortical wall had become attached to and incorporated into the ALL. Thus, the ALL seemed to act as an active periosteum for ossification. Although our materials were limited in number and stage, we hypothesized that, in contrast to the PLL, the mature anterior cortical wall corresponds to a calcified fibrocartilage adjacent to the ALL and forms a bone-ligament interface maintaining an ossification potential.