Matrix changes during long-term cultivation of cartilage, organoid or high-density cultures

Abstract

In high density (organoid or micromass) cultures of prechondrogenic mesenchymal cells from limb buds of 12-day-old mouse embryos typical cartilaginous tissue develops after 3 days. Immunomorphological investigations have shown that it contains the typical components of the cartilaginous matrix, such as collagen type 11 and cartilage-specific proteoglycans. After a 2-week cultivation period hypertrophic cartilage cells develop to an increasing extent. Many of these cells as well as normal chondroblasts detach from the matrix from the 2nd week in vitro onwards to assume a fibroblast-like appearance. At the same time thick (25-65 nm) collagenous fibrils occur at the surface of these cells. These thick fibrils contain collagen type I, as shown by immunomorphology. Hence, in these older cartilage cultures chondroblasts change their synthesis programme or direction of differentiation. Consequently, a model for the study of c~dedifferentiationn of cartilage and possibly also transformation of cartilage cells to osteoblasts has become available.