Journal of Cell Science, Vol 107, Issue 1 17-27, Copyright © 1994 by Company of Biologists

JOURNAL ARTICLES

Phenotypic stability of bovine articular chondrocytes after long-term culture in alginate beads

HJ Hauselmann, RJ Fernandes, SS Mok, TM Schmid, JA Block, MB Aydelotte, KE Kuettner and EJ Thonar
Department of Biochemistry, Rush Medical College, Rush-Presbyterian-St. Luke's Medical Center, Chicago, IL 60612.

Articular chondrocytes embedded in alginate gel produce de novo a matrix rich in collagens and proteoglycans. A major advantage of this culture system is that the cells can be recovered by chelating the calcium, which otherwise maintains the alginate in its gel state. Chondrocytes thus released are surrounded by tightly bound cell-associated matrix, which seems to correspond to the pericellular and territorial matrices identified in cartilage by electron microscopy. The cells and their associated matrix can be easily separated by mild centrifugation from more soluble matrix components derived principally from the 'interterritorial' matrix. This new cell culture system thus makes it possible to study the assembly and turnover of molecules present in two distinct matrix pools. Importantly, a significant proportion of the aggrecan molecules in each of these two pools can be extracted using a non-denaturing solvent, thereby making possible studies of the metabolism and turnover of native proteoglycan aggregates. We show in this report that chondrocytes isolated from the full depth of adult bovine articular cartilage and maintained for 8 months in alginate gel are still metabolically active and continue to synthesize cartilage-specific type II collagen and aggrecan. The cells did not synthesize large amounts of type I collagen or of the small nonaggregating proteoglycans as usually occurs when chondrocytes lose their phenotypic stability. After this extended period of time in culture, the cells were present as two populations exhibiting differences in size, shape and amount of extracellular matrix surrounding them. The first population was found only near the surface of the bead: these cells were flattened and surrounded by a matrix sparse in proteoglycans and collagen fibrils. The second population was found throughout the remaining depth of the bead: the cells were more round and almost always surrounded by a basket-like meshwork consisting of densely packed fibrils running tangential to the surface.

This article has been cited by other articles:

				A. J. Kvist, A. E. Johnson, M. Morgelin, E. Gustafsson, E. Bengtsson, K. Lindblom, A. Aszodi, R. Fassler, T. Sasaki, R. Timpl, et al. Chondroitin Sulfate Perlecan Enhances Collagen Fibril Formation: IMPLICATIONS FOR PERLECAN CHONDRODYSPLASIAS J. Biol. Chem., November 3, 2006; 281(44): 33127 - 33139. [Abstract] [Full Text] [PDF]

				M. Akmal, A. Anand, B. Anand, M. Wiseman, A. E. Goodship, and G. Bentley The culture of articular chondrocytes in hydrogel constructs within a bioreactor enhances cell proliferation and matrix synthesis J Bone Joint Surg Br, April 1, 2006; 88-B(4): 544 - 553. [Abstract] [Full Text] [PDF]

				J Wang, D Elewaut, I Hoffman, E M Veys, and G Verbruggen Physiological levels of hydrocortisone maintain an optimal chondrocyte extracellular matrix metabolism Ann Rheum Dis, January 1, 2004; 63(1): 61 - 66. [Abstract] [Full Text] [PDF]

				J. Kisiday, M. Jin, B. Kurz, H. Hung, C. Semino, S. Zhang, and A. J. Grodzinsky Self-assembling peptide hydrogel fosters chondrocyte extracellular matrix production and cell division: Implications for cartilage tissue repair PNAS, July 23, 2002; 99(15): 9996 - 10001. [Abstract] [Full Text] [PDF]

				Y.-M. Yoon, S.-J. Kim, C.-D. Oh, J.-W. Ju, W. K. Song, Y. J. Yoo, T.-L. Huh, and J.-S. Chun Maintenance of Differentiated Phenotype of Articular Chondrocytes by Protein Kinase C and Extracellular Signal-regulated Protein Kinase J. Biol. Chem., March 1, 2002; 277(10): 8412 - 8420. [Abstract] [Full Text] [PDF]

				M. G. Attur, M. N. Dave, M. Y. Leung, C. Cipolletta, M. Meseck, S. L. C. Woo, and A. R. Amin Functional Genomic Analysis of Type II IL-1{beta} Decoy Receptor: Potential for Gene Therapy in Human Arthritis and Inflammation J. Immunol., February 15, 2002; 168(4): 2001 - 2010. [Abstract] [Full Text] [PDF]

				S. Chubinskaya, K. Huch, M. Schulze, L. Otten, M. B. Aydelotte, and A. A. Cole Gene Expression by Human Articular Chondrocytes Cultured in Alginate Beads J. Histochem. Cytochem., October 1, 2001; 49(10): 1211 - 1220. [Abstract] [Full Text] [PDF]

				K F Almqvist, L Wang, J Wang, D Baeten, M Cornelissen, R Verdonk, E M Veys, and G Verbruggen Culture of chondrocytes in alginate surrounded by fibrin gel: characteristics of the cells over a period of eight weeks Ann Rheum Dis, August 1, 2001; 60(8): 781 - 790. [Abstract] [Full Text] [PDF]

				H. K. Ronday, W. H. van der Laan, P. P. Tak, J. A. D. M. de Roos, R. A. Bank, J. M. TeKoppele, C. J. Froelich, C. E. Hack, P. C. W. Hogendoorn, F. C. Breedveld, et al. Human granzyme B mediates cartilage proteoglycan degradation and is expressed at the invasive front of the synovium in rheumatoid arthritis Rheumatology, January 1, 2001; 40(1): 55 - 61. [Abstract] [Full Text] [PDF]

				R. S. SELLERS, R. ZHANG, S. S. GLASSON, H. D. KIM, D. PELUSO, D. A. D'AUGUSTA, K. BECKWITH, and E. A. MORRIS Repair of Articular Cartilage Defects One Year After Treatment with Recombinant Human Bone Morphogenetic Protein-2 (rhBMP-2) J. Bone Joint Surg. Am., February 1, 2000; 82(2): 151 - 60. [Abstract] [Full Text]

				Y. Nishida, C. B. Knudson, J. J. Nietfeld, A. Margulis, and W. Knudson Antisense Inhibition of Hyaluronan Synthase-2 in Human Articular Chondrocytes Inhibits Proteoglycan Retention and Matrix Assembly J. Biol. Chem., July 30, 1999; 274(31): 21893 - 21899. [Abstract] [Full Text] [PDF]

				H. J. Hauselmann, M. Stefanovic-Racic, B. A. Michel, and C. H. Evans Differences in Nitric Oxide Production by Superficial and Deep Human Articular Chondrocytes: Implications for Proteoglycan Turnover in Inflammatory Joint Diseases J. Immunol., February 1, 1998; 160(3): 1444 - 1448. [Abstract] [Full Text] [PDF]

				L. E. Freed, R. Langer, I. Martin, N. R. Pellis, and G. Vunjak-Novakovic Tissue engineering of cartilage in space PNAS, December 9, 1997; 94(25): 13885 - 13890. [Abstract] [Full Text] [PDF]

				R. M. Dharmavaram, G. Liu, S. D. Mowers, and S. A. Jimenez Detection and Characterization of Sp1 Binding Activity in Human Chondrocytes and Its Alterations during Chondrocyte Dedifferentiation J. Biol. Chem., October 24, 1997; 272(43): 26918 - 26925. [Abstract] [Full Text] [PDF]

				A. Rossi, J. Bonaventure, A.-L. Delezoide, G. Cetta, and A. Superti-Furga Undersulfation of Proteoglycans Synthesized by Chondrocytes from a Patient with Achondrogenesis Type 1B Homozygous for an L483P Substitution in the Diastrophic Dysplasia Sulfate Transporter J. Biol. Chem., August 2, 1996; 271(31): 18456 - 18464. [Abstract] [Full Text] [PDF]

				E Kolettas, L Buluwela, M. Bayliss, and H. Muir Expression of cartilage-specific molecules is retained on long-term culture of human articular chondrocytes J. Cell Sci., January 5, 1995; 108(5): 1991 - 1999. [Abstract] [PDF]

				C. Ghayor, J.-F. Herrouin, C. Chadjichristos, L. Ala-Kokko, M. Takigawa, J.-P. Pujol, and P. Galera Regulation of Human COL2A1 Gene Expression in Chondrocytes. IDENTIFICATION OF C-Krox-RESPONSIVE ELEMENTS AND MODULATION BY PHENOTYPE ALTERATION J. Biol. Chem., August 25, 2000; 275(35): 27421 - 27438. [Abstract] [Full Text] [PDF]

				M. G. Attur, M. Dave, C. Cipolletta, P. Kang, M. B. Goldring, I. R. Patel, S. B. Abramson, and A. R. Amin Reversal of Autocrine and Paracrine Effects of Interleukin 1 (IL-1) in Human Arthritis by Type II IL-1 Decoy Receptor. POTENTIAL FOR PHARMACOLOGICAL INTERVENTION J. Biol. Chem., December 15, 2000; 275(51): 40307 - 40315. [Abstract] [Full Text] [PDF]

				M. Demoor-Fossard, P. Galera, M. Santra, R. V. Iozzo, J.-P. Pujol, and F. Redini A Composite Element Binding the Vitamin D Receptor and the Retinoic X Receptor alpha Mediates the Transforming Growth Factor-beta Inhibition of Decorin Gene Expression in Articular Chondrocytes J. Biol. Chem., September 28, 2001; 276(40): 36983 - 36992. [Abstract] [Full Text] [PDF]