A suitable culture medium for ossification of embryonic chick femur in organ culture.

To establish a culture medium which allows ossification in organ culture, 9-day-old embryonic chick femurs were cultured in variously supplemented BGJb-HW2 media. Changes of Ca and Pi concentrations in the BGJb-HW2 medium or the 10% addition of chick embryo extract (CEE) did not induce ossification. Furthermore, combinations of the 10% CEE with a high Ca x Pi product or with 5 mM beta-glycerophosphate (beta-GP) or with 10% horse serum plus a high Ca x Pi product often caused pathological abnormalities in the periosteum. On the other hand, BGJb-HW2 medium supplemented with 5 mM beta-GP induced development of ossification. The Ca content of femurs and the diaphysial hydroxyproline content were markedly increased. Histological observation showed a formation of a thick and active periosteum, numerous osteoblastic cells, a sufficient amount of osteoid tissue and well developed calcified trabeculae without any pathological changes. Thus, the organ culture system using this medium was considered to be an appropriate one for studies on osteogenesis in vitro.