Ossification in tissue culture: I. Histological development of the femur of chick embryo in various liquid media

Abstract

By employing the roller-tube method without plasma, pair-mate cultivation of the femora of 9-day chick embryos was made in media, varied in composition and proportion, of horse serum (HS), chick embryo extract (CEE) and Gey's saline solution (GS). 1.1. When cultivated without CEE, elongation of the explants was significantly promoted in the above 30 per cent HS as compared with the control medium, CEE:HS:GS(1:2:7). Histologically, proliferation and hypertrophy of cartilage cells were always observed but the periosteum often disappeared.2.2. When cultivated without HS, successive growth, though inferior to the control, was observed in 10 per cent CEE. Histologically, cartilage cells were normal and the periosteum was thin but active.3.3. When cultivated in the media containing 10 per cent CEE and HS of various concentrations, elongation was accelerated in the order of HS 20% < 80% < 70% < 60% = 40% < 50%. Histologically, on employing the medium CEE:HS:GS(1:2:7), osteoid tissue was remarkably accumulated in osteogenic layer of the periosteum, and with increasing concentration of HS nearly physiologic ossification including calcification of the osteoid was obtained. In the above 60 per cent HS, however, granular calcium deposition was observed over the cartilage matrix at the end of diaphysis.