Long-term organ culture of embryonic chick femora: A system for investigating bone and cartilage formation at an intermediate level of organization

Abstract

Bone organ culture is an experimental system in which skeletal cells remain within their extracellular matrix but are removed from systemic influences. Femurs from 14-day-old chick embryos, which contain bone and cartilage matrix in approximately equal proportions, were cultured for up to 9 days in a serum-free medium. Cell proliferation, differentiation into chondrocytes and osteoblasts, formation of bone and cartilage matrix, and in vitro mineralization as well as bone and cartilage resorption were assessed using histologic and analytic methods. Particular attention was paid to the differences between cartilage and bone growth and to interpreting analytic data in the light of histologic observations. The first 2 days of culture represented an “adaptation” period, characterized by the release of intracellular enzymes into the culture medium, probably as a consequence of cell breakdown. Days 3–9 in culture represented a period of “steady growth” during which skeletal cells continued to multiply in the absence of fetal serum and to secrete large amounts of bone and cartilage matrix. De novo mineralization could be induced by Ca-ß-glycerophosphate, but calcium deposits in tissues other than bone and cartilage were also induced. Resorption of bone or cartilage matrix was virtually absent. Bone organ culture facilitates the study of bone and cartilage formation at an intermediate level of organization and thereby provides the necessary link between in vivo studies and investigations at the cellular level