Expression of the human chondrocyte phenotype in vitro
Rent the article at a discountRent now
* Final gross prices may vary according to local VAT.Get Access
We report a culture scheme in which human epiphyseal chondrocytes lose their differentiated phenotype in monolayer and subsequently reexpress the phenotype in an agarose gel. The scheme is based on a method using rabbit chondrocytes. Culture in monolayer allowed small quantities of cells to be amplified and provided a starting point to study expression of the differentiated human chondrocyte phenotype. The cells cultured in monolayer produced type I procollagen, fibronectin, and small noncartilaginous proteoglycans. Subsequent culture in agarose was associated with the acquisition of typical chondrocyte ultrastructural features and the synthesis of type II collagen and cartilage-specific proteoglycans. The switch from the nonchondrocyte to the differented chondrocyte phenotype occurred under these conditions between 1 and 2 wk of agarose culture and was not necessarily homogeneous throughout a culture. This culture technique will facilitate direct investigation of human disorders of cartilage that have been addressed in the past by alternative approaches.
- Abbott, J.; Holtzer, H. The loss of phenotypic traits by differentiated cells. III. The reversible behavior of chondrocytes in primary culture. J. Cell Biol. 28:473–487; 1966. CrossRef
- Aydelotte, M. B.; Kuettner, K. E. Differences between subpopulations of culture bovine articular chondrocytes, I. morphology and cartilage matrix production. Connect Tissue Res. 18:205–222; 1988.
- Bassleer, C.; Gysen, P.; Foidart, J. M., et al. Human chondrocytes in tridimensional culture. In Vitro 22:113–119; 1986.
- Benya, P. D.; Shaffer, J. D. Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell 30:215–224; 1982. CrossRef
- Capasso, O.; Gionti, E.; Pontarilli, G., et al. The culture of chick embryo chondrocytes and the control of their differentiated functions in vitro. I. Characterization of the chondrocyte-specific phenotypes. Exp. Cell Res. 142:197–206; 1982. CrossRef
- Castagnola, P.; Moro, G.; Descalze-Cancedda, F., et al. Type X collagen synthesis during in vitro development of chick embryo tibial chondrocytes. J. Cell Biol. 102:2310–2317; 1986. CrossRef
- Ceylon, S.; Abbott, J.; Holtzer, S., et al. The loss of phenotypic traits by differentiated cells. VI. Behavior of the progeny of a single chondrocyte. J. Exp. Med. 130:417–442; 1969. CrossRef
- Coon, H. G. Clonal stability and phenotypic expression of chick cartilage cells in vitro. Proc. Natl. Acad. Sci. USA 55:66–73; 1966. CrossRef
- Delbruck, A.; Dresow, B.; Gurr, E., et al. In vitro culture of human chondrocytes from adult subjects. Conn. Tissue Res. 15:115–172; 1986. CrossRef
- Deshmukh, K.; Kline, W. G. Characterization of collagen and its precursors synthesized by rabbit-articular-cartilage cells in various culture systems. Eur. J. Biochem. 69:117–123; 1976. CrossRef
- Glowacki, J.; Trepman, E.; Folkman J. Cell shape and phenotypic expression in chondrocytes. Proc. Soc. Exp. Biol. Med. 172:93–98; 1983.
- Holtzer, H.; Abbott, J.; Lash, J., et al. The loss of phenotypic traits by differentiated cells in vitro, I. Dedifferentiation of cartilage cells. Proc. Natl. Acad. Sci. USA 46:1533–1542; 1960. CrossRef
- Horton, W. A.; Machado, M. M. Extracellular matrix alterations during endochondral ossification in humans. J. Orthop. Res. 6:793–803; 1988. CrossRef
- Horwitz, A. L.; Dorfman, A. The growth of cartilage cells in soft agar and liquid suspension. J. Cell Biol. 45:434–438; 1970. CrossRef
- Johnson, G. D.; Nagueira Araugo, G. M. de C. A simple method of reducing the fading of immunofluroescence during microscopy. J. Immunol. Methods 43:349–350; 1981. CrossRef
- Kuettner, K. E.; Pauli, B. U.; Gall, G., et al. Synthesis of cartilage matrix by mammalian chondrocytes in vitro. I. isolation, culture characteristics, and morphology. J. Cell Biol. 93:743–750; 1982. CrossRef
- Kuettner, K. E.; Memoli, V. A.; Pauli, B. U., et al. Synthesis of cartilage matrix by mammalian chondrocytes in vitro. II. Maintenance of collagen and proteoglycan phenotype. J. Cell Biol. 93:751–757; 1982. CrossRef
- Layman, D. L.; Sokoloff, L.; Miller, E. J. Collagen synthesis by articular chondrocytes in monolayer culture. Exp. Cell Res. 73:107–112; 1972. CrossRef
- Maor, G.; Von Der Mark, K.; Reddi, H., et al. Acceleration of cartilage and bone differentiation on collagenous substrata. Collagen Rel. Res. 7:351–370; 1987.
- Manning, W. K.; Bonner, W. M., Jr. Isolation and culture of chondrocytes from human adult articular cartilage. Arthritis and Rheumatism. 10:235–239; 1967. CrossRef
- Norby, D. P.; Malemud, C. J.; Sokoloff, L. Differences in the collagen types synthesized by lapine articular chondrocytes in spinner and monolayer culture. Arthritis Rheum. 20:709–716; 1977. CrossRef
- Prockop, D. J.; Pettengill, O.; Holtzer, H. Incorporation of sulfate and the synthesis of collagen by cultures of embryonic chondrocytes. Biochim. Biophys. Acta. 83:189–196; 1964.
- Roughley, P. J.; White, R. J. Age related changes in the structure of the proteoglycan subunits from human articular cartilage. J. Biol. Chem. 255:217–224; 1980.
- Sokoloff, L.; Malemud, C. J.; Srivastava, V. M. L., et al. In vitro culture of articular chondrocytes. Fed. Proc. 32:1499–1502; 1973.
- Solursh, M.; Jensen, K. L.; Reiter, R. S., et al. Environmental regulation of type X collagen production by cultures of limb mesenchyme, mesectoderm, and sternal chondrocytes. Dev. Biol. 117:90–101; 1986. CrossRef
- Sun, D.; Aydelotte, M. B.; Maldanado, B., et al. Clonal analysis of the population of chondrocytes from the swarm rat chondrosarcoma in agarose culture. J. Orthop. Res. 4:427–436; 1986. CrossRef
- Tacchetti, C.; Quarto, R.; Nitsch, L., et al. In vitro morphogenesis of chick embryo hypertrophic cartilage. J. Cell Biol. 105:999–1006; 1987. CrossRef
- Wiebkin, O. W.; Muir, H. Synthesis of cartilage-specific proteoglycans by suspension cultures of adult chondrocytes. Biochem. J. 164:269–272; 1977.
- Wiebkin, O. W.; Muir, H. Synthesis of proteoglycans by suspension and monolayer cultures of adult chondrocytes and de novo cartilage nodules. The effect of hyaluronic acid. J. Cell Sci. 27:199–211; 1977.
- Expression of the human chondrocyte phenotype in vitro
In Vitro Cellular & Developmental Biology
Volume 25, Issue 7 , pp 659-668
- Cover Date
- Print ISSN
- Online ISSN
- Additional Links
- human chondrocyte
- agarose culture
- Industry Sectors