Synthesis of human cartilage using organotypic cell culture

The limited supply of fresh autologous cartilage tissue for use in reconstructive surgery necessitates the use of vital banked allografts. A feasible in vitro production of cartilage tissue composed of living cells requires the use of modern tissue culture techniques retaining the phenotypic characteristics of chondrocytes. With this purpose in mind, human chondrocytes were isolated and cultured using different culture procedures: monolayer, suspension and agar gel. The differentiation state of chondrocytes as well as proteoglycan and collagen syntheses were assessed by histochemical and immunohistochemical methods. Whereas chondrocytes in monolayer displayed an unstable phenotype and tended to dedifferentiate, in three-dimensional culture the chondrocytes remained morphologically, phenotypically and functionally differentiated. Furthermore, an accumulation of matrix products pericellularly was observed in the agar gel. The results suggest that three-dimensional cultures in agar gel may allow the in vitro production of bioartificial cartilage for transplantation.