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Human Cell Culture Protocols pp 301–336Cite as

Human Chondrocyte Cultures as Models of Cartilage-Specific Gene Regulation

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 806))

Abstract

The human adult articular chondrocyte is a unique cell type that has reached a fully differentiated state as an end point of development. Within the cartilage matrix, chondrocytes are normally quiescent and maintain the matrix constituents in a low-turnover state of equilibrium. Isolated chondrocytes in culture have provided useful models to study cellular responses to alterations in the environment such as those occurring in different forms of arthritis. However, expansion of primary chondrocytes in monolayer culture results in the loss of phenotype, particularly if high cell density is not maintained. This chapter describes strategies for maintaining or restoring differentiated phenotype by culture in suspension, gels, or scaffolds. Techniques for assessing phenotype involving primarily the analysis of synthesis of cartilage-specific matrix proteins as well as the corresponding mRNAs are also described. Approaches for studying gene regulation, including transfection of promoter-driven reporter genes with expression vectors for transcriptional and signaling regulators, chromatin immunoprecipitation, and DNA methylation are also described.

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Acknowledgments

Dr. Goldring’s research related to this project was supported in part by grants from the National Institutes of Health, AG022021, AR054887, and AR045378, and the Arthritis Foundation. The authors are grateful to Lujian Tan, Haibing Peng, Lii-Fang Suen, James Birkhead, Merrilee Flannery, James Robbins, Makoto Osaki, and Bob Choy for supplying technical expertise and data, and to Dr. Thomas Sculco at The Hospital for Special Surgery and Dr. Benjamin Bierbaum at the New England Baptist Hospital for supplying cartilage samples. We also wish to acknowledge Dr. Helmtrud (Trudy) I. Roach, recently deceased, for her contributions to our understanding of epigenetics in chondrocytes and for transferring the technology to our laboratory.

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  1. Laboratory for Cartilage Biology, The Hospital for Special Surgery, Weill Cornell Medical College, New York, NY, USA

    Miguel Otero, Marta Favero, Cecilia Dragomir, Karim El Hachem, Ko Hashimoto, Darren A. Plumb & Mary B. Goldring

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  1. Miguel Otero
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  2. Marta Favero
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Correspondence to Mary B. Goldring .

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  1. Institute of Liver Studies, King’s College Hospital, Denmark Hill, London, SE5 9RS, United Kingdom

    Ragai R. Mitry

  2. School of Medicine, Inst. Liver Studies, King's College London, Bessemer Rd., London, SE5 9RS, United Kingdom

    Robin D. Hughes

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Otero, M. et al. (2012). Human Chondrocyte Cultures as Models of Cartilage-Specific Gene Regulation. In: Mitry, R., Hughes, R. (eds) Human Cell Culture Protocols. Methods in Molecular Biology, vol 806. Humana Press. https://doi.org/10.1007/978-1-61779-367-7_21

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