Abstract
The chondrocyte is responsible for synthesis of cartilage matrix proteins, and, thereby, the specialized mechanical properties of articular cartilage, including tensile strength and resistence to mechanical loading (1). The limited repair response by chondrocytes accounts for a major component of the loss of articular cartilage in joint diseases such as osteoarthritis, a progressive disease associated with normal wear and tear of joints, aging, or trauma. Although research has been directed primarily toward developing therapeutic strategies that prevent degradation of cartilage matrix, recent work has also focused on promoting cartilage repair. Success of either strategy depends on the development of reliable cell culture models of human origin.
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References
Poole, A. R. (1993) Cartilage in health and disease, in Arthritis and Allied Conditions: A Textbook of Rheumatology (McCarty, D. J. and Koopman, W. P., eds.), Lea and Febiger, Philadelphia, pp. 279ā333.
Goldring, M. B. (1993) Degradation of articular cartilage in culture: regulatory factors, in Joint Cartilage Degradation: Basic and Clinical Aspects (Woessner, J. F., Jr. and Howell, D. S., eds.), Marcel Dekker, New York, pp. 281ā345.
Goldring, M. B., Sandell, L. J., Stephenson, M. L., and Krane, S. M. (1986) Immune interferon suppresses levels of procollagen mRNA and type II collagen synthesis in cultured human articular and costal chondrocytes. J. Biol. Chem. 261, 9049ā9056.
Goldring, M. B. and Krane, S. M. (1987) Modulation by recombinant interleukin 1 of synthesis of types I and III collagens and associated procollagen mRNA levels in cultured human cells. J. Biol. Chem. 262, 16,724ā16,729.
Goldring, M. B., Birkhead, J., Sandell, L. J., Kimura, T., and Krane, S. M. (1988) Interleukin 1 suppresses expression of cartilage-specific types II and IX collagens and increases types I and III collagens in human chondrocytes. J. Clin. Invest. 82, 2026ā2037.
von der Mark, K., Gauss, V., von der Mark, H., and Muller, P. (1977) Relationship between cell shape and type of collagen synthesised as chondrocytes lose their cartilage phenotype in culture. Nature 267, 531ā532.
Benya, P. D. and Shaffer, J. D. (1982) Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell 30, 215ā224.
Castagnola, P., Moro, G., Descalzi-Cancedda, F., and Cancedda, R. (1986) Type X collagen synthesis during in vitro development of chick embryo tibial chondrocytes. J. Cell Biol. 102, 2310ā2317.
Alema, S., Tato, F., and Boettiger, D. (1985) Myc and src oncogenes have complementary effects on cell proliferation and expression of specific extracellular matrix components in definitive chondroblasts. Mol. Cell. Biol. 5, 538ā544.
Gionti, E., Pontarelli, G., and Cancedda, R. (1985) Avian myelocytomatosis virus immortalizes differentiated quail chondrocytes. Proc. Natl. Acad. Sci. USA 82, 2756ā2760.
Horton, W. E., Jr., Cleveland, J., Rapp, U., Nemuth, G., Bolander, M., Doege, K., Yamada, Y., and Hassell, J. R. (1988) An established rat cell line expressing chondrocyte properties. Exp. Cell Res. 178, 457ā468.
Thenet, S., Benya, P. D., Demignot, S., Feunteun, J., and Adolphe, M. (1992) SV40-immortalization of rabbit articular chondrocytes: alteration of differentiated functions. J. Cell. Physiol. 150, 158ā167.
Mallein-Gerin, F. and Olsen, B. R. (1993) Expression of simian virus 40 large T (tumor) oncogene in chondrocytes induces cell proliferation without loss of the differentiated phenotype. Proc. Natl. Acad. Sci. USA 90, 3289ā3293.
Benoit, B., Thenet-Gauci, S., Hoffschir, F., Penformis, P., Demignot, S., and Adolphe, M. (1995) SV40 large T antigen immortalization of human articular chondrocytes. In Vitro Cell. Dev. Biol. 31, 174ā177.
Mataga, N., Tamura, M., Yanai, N., Shinomura, T., Kimata, K., Obinata, M., and Noda, M. (1996) Establishment of a novel chondrocyte-like cell line derived from transgenic mice harboring the temperature-sensitive simian virus 40 large T-antigen. J. Bone Miner. Res. 11, 1646ā1654.
Lefebvre, V., Garofalo, S., and deCrombrugghe, B. (1995) Type X collagen gene expression in mouse chondrocytes immortalized by a temperature-sensitive simian virus 40 large tumor antigen. J. Cell Biol. 128, 239ā245.
Goldring, M. B., Birkhead, J. R., Suen, L.-F., Yamin, R., Mizuno, S., Glowacki, J., Arbiser, J. L., and Apperley, J. F. (1994) Interleukin-1 Ī²-modulated gene expression in immortalized human chondrocytes. J. Clin. Invest. 94, 2307ā2316.
Fanning, E. and Knippers, R. (1992) Structure and function of simian virus 40 large T antigen. Annu. Rev. Biochem. 61, 55ā85.
Duncan, E. L., Whitaker, N. J., Moy, E. L., and Reddel, R. R. (1993) Assignment of SV40-immortalized cells to more than one complementation group for immortalization. Exp. Cell Res. 205, 337ā344.
Rowe, D. W., Moen, R. C, Davidson, J. M., Byers, P. H., Bornstein, P., and Palmiter, R. D. (1978) Correlation of procollagen mRNA levels in normal and transformed chick embryo fibroblasts with different rates of procollagen synthesis. Biochemistry 17, 1581ā1590.
Berman, A. E. and Morozevich, G. E. (1990) Secretion and intracellular degradation of collagen in cultures of normal and SV-40-transformed human fibroblasts. FEBS Lett. 263, 285ā263.
Woodworm, C. D., Kreider, J. W., Mengel, L., Miller, T., Meng, Y. L., and H. C, I. (1988) Tumorigenicity of simian virus 40-hepatocyte cell lines: effect of in vitro and in vivo passage on expression of liver-specific genes and oncogenes. Mol. Cell Biol. 8, 4492ā4501.
Iwamoto, M., Yagami, K., LuValle, P., Olsen, B. R., Petropoulos, C. J., Ewert, D. L., and Pacifici, M. (1993) Expression and role of c-myc in chondrocytes undergoing endochondral ossification. J. Biol. Chem. 268, 9645ā9652.
Jat, P. S. and Sharp, P. A. (1989) Cell lines established by a temperature-sensitive simian virus 40 large-T-antigen gene are growth restricted at the nonpermissive temperature. Mol. Cell. Biol. 9, 1672ā1681.
Mitchell, P. J., Wang, C., and Tjian, R. (1987) Positive and negative regulation of transcription in vitro: enhancer binding protein AP-2 is inhibited by SV40 T antigen. Cell 50, 847ā861.
Hansell, E. J., Frisch, S. M., Tremble, P. M., Murnane, J. P., and Werb, Z. (1995) Simian virus 40 transformation alters the actin cytoskeleton, expression of matrix metalloproteinases and inhibitor of metalloproteinases, and invasive behavior of human skin fibroblasts. Biochem. Cell Biol. 73, 373ā389.
Logan, S. K., Hansell, E. J., Damsky, C. H., and Werb, Z. (1996) T-Antigen inhibits metalloproteinase expression and invasion in human placental cells transformed with temperature-sensitive simian virus 40. Matrix Biol. 15, 81ā89.
Glowacki, J., Trepman, E., and Folkman, J. (1983) Cell shape and phenotypic expression in chondrocytes. Proc. Soc. Exp. Biol. Med. 172, 93ā98.
Reginato, A. M., Iozzo, R. V., and Jimenez, S. A. (1994) Formation of nodular structures resembling mature articular cartilage in long-term primary cultures of human fetal epiphyseal chondrocytes on hydrogel substrate. Arthritis Rheum. 37, 1338ā1349.
Paulsen, D. F. and Solursh, M. (1988) Microtiter micromass cultures of limb-bud mesenchymal cells. In Vitro Cell. Dev. Biol. 24, 138ā147.
Kato, Y., Iwamoto, M., Koike, T., Suzuki, F., and Takano, Y. (1988) Terminal differentiation and calcification in rabbit chondrocyte cultures grown in centrifuge tubes: Regulation by transforming growth factor Ī² and serum factors. Proc. Natl. Acad. Sci. USA 85, 9552ā9556.
Ballock, R. T. and Reddi, A. H. (1994) Thyroxine is the serum factor that regulates morphogenesis of columnar cartilage from isolated chondrocytes in chemically defined medium. J. Cell Biol. 126, 1311ā1318.
Denker, A. E., Nicoll, S. B., and Tuan, R. S. (1995) Formation of cartilage-like spheroids by micromass cultures of murine C3H10T1/2 cells upon treatment with transforming growth factor-Ī²1. Differentiation 59, 25ā34.
Sun, S., Aydelotte, M. B., Maldonaldo, B., Kuettner, K. E., and Kimura, J. H. (1986) Clonal analysis of the population of chondrocytes from the Swarm rat chondrosarcoma in agarose culture. J. Orthopaed. Res. 4, 427ā436.
Guo, J., Jourdian, G. W., and MacCallum, D. K. (1989) Culture and growth characteristics of chondrocytes encapsulated in alginate beads. Connect. Tiss. Res. 19, 277ā297.
Hauselmann, H. J., Fernandes, R. J., Mok, S. S., Schmid, T. M., Block, J. A., Aydelotte, M. B., Kuettner, K. E., and Thonar, E. J. (1994) Phenotypic stability of bovine articular chondrocytes after long-term culture in alginate beads. J. Cell Sci. 107, 17ā27.
Gibson, G. J., Schor, S. L., and Grant, M. E. (1982) Effects of matrix macromol-ecules on chondrocyte gene expression: synthesis of a low molecular weight collagen species by cells cultured within collagen gels. J. Cell Biol. 93, 767ā774.
Freed, L. E., Marquis, J. C, Nohria, A., Emmanual, J., Mikos, A. G., and Langer, R. (1993) Neocartilage formation in vitro and in vivo using cells cultured on synthetic biodegradable polymers. J. Biomed. Mater. Res. 27, 11ā23.
Nicoll, S. B., Denker, A. E., and Tuan, R. S. (1995) In vitro characterization of transforming growth factor-Ī²1-loaded composites of biodegradable polymer and mesenchymal cells. Cells Materials 5, 231ā244.
Mizuno, S. and Glowacki, J. (1996) Chondroinduction of human dermal fibroblasts by demineralized bone in three-dimensional culture. Exp. Cell Res..
Goldring, M. B. (1996) Human chondrocyte cultures as models of cartilage-specific gene regulation, in Methods in Molecular Biology: Human Cell Culture Protocols (Jones, G. E., ed.), Humana, Totawa, NJ, pp. 217ā231.
Sams, A. E. and Nixon, A. J. (1995) Chondrocyte-laden collagen scaffolds for resrufacing extensive articular cartilage defects. Osteoarthritis Cartilage 3, 47ā59.
Brittberg, M., Lindahl, A., Nilsson, A., Ohlsson, C, Isaksson, O., and Peterson, L. (1994) Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. N. Engl. J. Med. 331, 889ā895.
Crystal, R. G. (1995) Transfer of genes to humans: Early lessons and obstacles. Science 270, 404ā410.
Bandara, G., Mueller, G. M., Galea-Lauri, J., Tindal, M. H., Georgescu, H. I., Sucharek, M. K., et al. (1993) Intraarticular expression of the interleukin-1 receptor antagonist protein by ex vivo gene transfer. Proc. Natl. Acad. Sci. USA 90, 10,764ā10,768.
Geiler, T., Kriegsmann, J., Keyszer, G. M., Gay, R. E., and Gay, S. (1994) A new model for rheumatoid arthritis generated by engraftment of rheumatoid synovial tissue and normal human cartilage into SCID mice. Arthritis Rheum. 37, 1664ā1671.
Harlow, E., Crawford, L. V., Pirn, D. C., and Williamson, N. M. (1981) Monoclonal antibodies specific for simian virus 40 tumor antigens. J. Virol. 39, 861ā869.
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680ā685.
Miller, A. D. and Buttimore, C. (1986) Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production. Mol. Cell Biol. 6, 2895ā2902.
Vogel, K. G., Sandy, J. D., Pogany, G., and Robbins, J. R. (1994) Aggrecan in bovine tendon. Matrix Biol. 14, 171ā179.
Robbins, J. R. and Vogel, K. G. Mechanical loading and TGF-Ī² regulate proteoglycan synthesis in tendon. Arch. Biochem. Biophys. submitted.
Lefebvre, V., Garofalo, S., Zhou, G., Metsaranta, M., Vuorio, E., and deCrombrugghe, B. (1994) Characterization of primary cultures of chondrocytes from type II collagen/Ī²-galactosidase transgenic mice. Matrix Biol. 14, 329ā335.
Lum, Z.-P., Hakala, B. E., Mort, J. S., and Recklies, A. D. (1996) Modulation of the catabolic effects of interleukin-1Ī²3 on human articular chondrocytes by transforming growth factor-Ī². J. Cell. Physiol. 166.
Bonaventure, J., Kadhom, N., Cohen-Solal, L., Ng, K. H., Bourguigno, J., Lasselin, C., and Freisinger, P. (1994) Reexpression of cartilage-specific genes by dedifferentiated human articular chondrocytes cultured in alginate beads. Exp. Cell Res. 212, 97ā104.
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Robbins, J.R., Goldring, M.B. (1999). Preparation of Immortalized Human Chondrocyte Cell Lines. In: Morgan, J.R., Yarmush, M.L. (eds) Tissue Engineering Methods and Protocols. Methods in Molecular Medicineā¢, vol 18. Humana Press. https://doi.org/10.1385/0-89603-516-6:173
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DOI: https://doi.org/10.1385/0-89603-516-6:173
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