Confocal analysis of the molecular heterogeneity in the pericellular microenvironment produced by adult canine chondrocytes cultured in agarose gel

摘要: Adult articular chondrocytes are each surrounded by a heterogeneous microenvironment and together form the chondron. Since little is known of chondron development, agarose gel culture, confocal immunohistochemistry image analysis have been used to characterize molecular anatomy temporal development chondrocyte pericellular in vitro. Two structurally distinct domains were identified during 12-week culture period. The first comprised narrow glycocalyx, 1–3 ·m width, which consolidated over time was rich collagen types II, VI, IX XI, fibronectin, decorin aggrecan epitopes, 5D4 HABR. second region emerged after 4–6 weeks progressively developed broad territorial up 12 wide around glycocalyx. Co-localization studies confirmed dominance epitopes 2B6, EFG-4, HABR domain, whereas surface density mapping with NIH revealed two patterns staining, one punctate stippled, other more uniform distribution. differentiation appeared analogous chondrons adult cartilage, provides an appropriate vitro model for further cell receptor function orchestration matrix assembly