Abstract
Differentiation of cartilage and chondro-osseous tissue occurs from a mesenchymal cell outgrowth in 88.4% of cultures of muscle tissue from near term rat fetuses when the following physico-chemical and nutritional conditions are present. The muscle is placed in a vessel made from demineralized rat bone matrix wetted with a plasma, which serves as a bridge between the explant and the substratum. The matrix serves as both a substratum and the delimiting area of the proliferating edge of the culture. When contact is made with the substratum, a chondroosseous morphogenetic pattern is imposed upon mesenchymal cells and their progeny. The chemically-defined medium 199 is used for the first 5 days to produce abundant outgrowth of mesenchyma which precedes all skeletal tissue differentiation. Medium 199 is replaced with the modified BGJ medium for 6 to 30 days to provide aeration, amino acids and vitamins in optimum quantities for synthesis of new cartilage matrix. High oxygen tension in the atmosphere surrounding the culture media is toxic to young mesenchymal cells and must be avoided during the initial phases of skeletal tissue development.
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