Bioassay of chondrocyte differentiation by bone morphogenetic protein

For sensitive bioassay of cartilage development in response to bone morphogenetic protein (BMP), the supernatant of interstitial fluid containing crud BMP was obtained from an implanted diffusion chamber, applied to a cellulose acetate film, and stained with Alcian blue. The optical density of the Alcian blue-stained spot was measured by a densitometer for quantitative analysis. Density of the Alcian blue-stained spot was markedly increased when cartilage developed outside the chamber. Occurrence of cartilage-specific proteoglycan-H (PG-H) and Type II collagen was seen in this intensely stained spot by dot-blot analysis. Electron microscopic examination revealed that the residue of interstitial fluid in the implanted diffusion chamber contained many collagen fibrils and proteoglycan granules.