The cultures were allowed to incorporate 35SO2-4 for various periods of time. 35S-labelled macromolecules were isolated from the medium, a trypsin digest of the cells and the cell residue. Ion-exchange chromatography separated the radioactive polysaccharides into heparan sulphate and a galactosaminoglycan population. Most heparan sulphate was in the trypsin digest and cell residue fractions. The galactosaminoglycan fractions were investigated by differential degradations with chondroitinase ABC and AC and ethanol fractionation. The medium galactosaminoglycans contained both glucuronic and iduronic acid residues and existed in copolymeric structures as chondroitin sulphate/dermatan sulphate hybrid molecules. Dermatan sulphate was also detected. In contrast, the trypsin-digest fraction contained mainly chondroitin sulphate-like molecules.