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Cell Cycle Synchronization of Primary and Cultured Articular Chondrocytes

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Cell-Cycle Synchronization

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2579))

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Abstract

Cell cycle synchronization allows cells in a culture, originally at different stages of the cell cycle, to be brought to the same phase. It is normally performed by applying cell cycle arresting chemical agents to cells cultured in monolayer. While effective, isolated chondrocytes tend to dedifferentiate when cultured in monolayer and typically require 3D culturing methods to ensure phenotypic stability. Here, we describe both the conventional cell cycle synchronization method for cells in monolayer culture and an adapted method of synchronizing primary chondrocytes directly during the cell isolation process to limit potential dedifferentiation. Different methods including serum-starvation and treatment with thymidine, nocodazole, aphidicolin, and RO-3306 can synchronize the chondrocytes at different discrete phases. A cell purity of more than 90% in the S phase can be achieved with simultaneous cell isolation and synchronization using double thymidine treatment, generating a population of synchronized chondrocytes that show increased matrix synthesis when subsequently cultured in 3D.

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Authors and Affiliations

  1. Department of Chemical Engineering, Ryerson University, Toronto, ON, Canada

    Loraine L. Y. Chiu, Omar D. Subedar & Stephen D. Waldman

  2. Li Ka Shing Knowledge Institute, St. Michael’s Hospital, Toronto, ON, Canada

    Loraine L. Y. Chiu, Omar D. Subedar & Stephen D. Waldman

Authors
  1. Loraine L. Y. Chiu
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  2. Omar D. Subedar
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  3. Stephen D. Waldman
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Corresponding author

Correspondence to Stephen D. Waldman .

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Editors and Affiliations

  1. Department of Medical Genetics, University of Alberta, Edmonton, AB, Canada

    Zhixiang Wang

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Chiu, L.L.Y., Subedar, O.D., Waldman, S.D. (2022). Cell Cycle Synchronization of Primary and Cultured Articular Chondrocytes. In: Wang, Z. (eds) Cell-Cycle Synchronization. Methods in Molecular Biology, vol 2579. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2736-5_9

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  • DOI: https://doi.org/10.1007/978-1-0716-2736-5_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2735-8

  • Online ISBN: 978-1-0716-2736-5

  • eBook Packages: Springer Protocols

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