title: Characterization of subpopulated articular chondrocytes separated by Percoll density gradient.
abstract: The aim of this study was to develop a method for fractionation of articular chondrocytes from the entire thickness of the tissue. Isolated chondrocytes from rabbit articular cartilage fractionated by centrifugation in a discontinuous Percoll gradient resulted in four cell fractions with two differing properties. The lowest-density fraction consisted mainly of large cells with small nuclei proliferated actively, maintained the chondrocytic phenotype, and secreted larger amounts of proteoglycan. In contrast, the highest-density fraction consisted of small cells with large nuclei proliferated slowly, did not express the chondrocytic phenotype, and produced larger amounts of interleukin 1-induced nitric oxide. Comparing our results with other previous reports, we find that fraction 1 cells are likely originated from the deep layer of the articular cartilage, whereas fraction 4 cells are tentatively categorized as chondrocytes from the superficial layer of cartilage. Centrifugal fractionation of articular chondrocytes via Percoll density gradient permits clear separation of these heterogeneous cells into different phenotypic populations and allows distinguishing of cells from the different layers of articular cartilage. This simple novel method will provide ready separation of articular chondrocytes for the investigation of the pathogenesis of articular cartilage.
authors.affiliation: Department of Orthopedic Surgery, Ajou University School of Medicine, Suwon, Kyeongki-Do, South Korea.
authors.forename: Byoung-Hyun
authors.initials: BH
authors.lastname: Min
authors.forename: Hyeon Joo
authors.initials: HJ
authors.lastname: Kim
authors.forename: Hanjo
authors.initials: H
authors.lastname: Lim
authors.forename: So Ra
authors.initials: SR
authors.lastname: Park
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002358:Cartilage, Articular / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism / Q000648:ultrastructure; D002455:Cell Division; D002469:Cell Separation; D019902:Chondrocytes / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D008854:Microscopy, Electron; D009569:Nitric Oxide / Q000096:biosynthesis; D011205:Povidone; D011509:Proteoglycans / Q000096:biosynthesis; D011817:Rabbits; D012822:Silicon Dioxide
chemical_list: D011509:Proteoglycans; D009569:Nitric Oxide; C016039:Percoll; D012822:Silicon Dioxide; D000469:Alkaline Phosphatase; D011205:Povidone