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Physioxic Culture of Chondrogenic Cells

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Cartilage Tissue Engineering

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2598))

Abstract

Cartilage resides under a low oxygen tension within articulating joints. The oxygen tension within cartilage of the knee joint has been measured to be between 2% and 5% oxygen. Although the literature has historically termed this level of oxygen as hypoxia, particularly when doing experiments in vitro in this range, this is actually the physiological oxygen tension experienced in vivo and is more accurately termed physioxia. In general, culture of chondrogenic cells under physioxia has demonstrated a donor-dependent beneficial effect on chondrogenesis, with an upregulation in cartilage genes (SOX9, COL2A1, ACAN) and matrix deposition (sulfated glycosaminoglycans (sGAGs), collagen II). Physioxia also reduces the expression of hypertrophic markers (COL10A1, MMP13). This chapter will outline the methods for the expansion and differentiation of chondrogenic cells under physioxia using oxygen-controlled incubators and glove box environments, with the typical assays used for qualitative and quantitative assessment of chondrogenesis.

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Acknowledgements

This work was supported by the DFG Research network (FOR2407/1): ExCarBon, the European Union Horizon 2020 research and innovation program (EU No.: 814444 (MEFISTO), and a Shriners Hospitals for Children research grant.

Author information

Authors and Affiliations

  1. Experimental Trauma Surgery, Department of Trauma Surgery, University Medical Center of Regensburg, Regensburg, Germany

    Girish Pattappa & Denitsa Docheva

  2. Department of Orthopaedics & Rehabilitation, Oregon Health & Science University, Portland, OR, USA

    Brandon D. Markway & Brian Johnstone

  3. Department of Musculoskeletal Tissue Regeneration, Orthopaedic Hospital König-Ludwig-Haus, University of Wurzburg, Wurzburg, Germany

    Denitsa Docheva

Authors
  1. Girish Pattappa
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  2. Brandon D. Markway
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  3. Denitsa Docheva
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  4. Brian Johnstone
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Corresponding author

Correspondence to Brian Johnstone .

Editor information

Editors and Affiliations

  1. AO Research Institute Davos, Davos Platz, Switzerland

    Martin J. Stoddart

  2. AO Research Institute Davos, Davos Platz, Switzerland

    Elena Della Bella

  3. UCB Pharma, Slough, UK

    Angela R. Armiento

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Pattappa, G., Markway, B.D., Docheva, D., Johnstone, B. (2023). Physioxic Culture of Chondrogenic Cells. In: Stoddart, M.J., Della Bella, E., Armiento, A.R. (eds) Cartilage Tissue Engineering. Methods in Molecular Biology, vol 2598. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2839-3_5

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  • DOI: https://doi.org/10.1007/978-1-0716-2839-3_5

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2838-6

  • Online ISBN: 978-1-0716-2839-3

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