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TGF-β1- and CCN2-Stimulated Sirius Red Assay for Collagen Accumulation in Cultured Cells

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CCN Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1489))

Abstract

A simple method for the determination of relative levels of insoluble collagen accumulation in fibroblast cultures is presented. Confluent cell cultures are provided with sodium ascorbate which is then permissive for collagen deposition. At intervals, cultures are fixed and stained successively with sirius red and then crystal Violet to, respectively, assess for relative changes in collagen accumulation in response to factors such as TGF-β1 or matricellular CCN2 and changes in DNA content as an index of changes in cell density.

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Acknowledgements

Edwin C. K. Heng and Vianca E. Cabrera are acknowledged for their help to optimize the Sirius Red method for determination of collagen accumulation by fibroblast cultures. This work was supported by NIH/NIDCR R01 DE11004.

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Authors and Affiliations

  1. Department of Molecular and Cell Biology, Henry M. Goldman School of Dental Medicine, Boston University, 700 Albany Street, W-201, Boston, MA, 02118, USA

    Philip C. Trackman Ph.D., Debashree Saxena & Manish V. Bais

Authors
  1. Philip C. Trackman Ph.D.
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  2. Debashree Saxena
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  3. Manish V. Bais
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Corresponding author

Correspondence to Philip C. Trackman Ph.D. .

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Editors and Affiliations

  1. Advanced Research Center for Oral and Craniofacial Sciences, Okayama University Dental School/Graduate School of Medicine Dentistry and Pharmaceutical Sciences, Okayama, Japan

    Masaharu Takigawa

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Trackman, P.C., Saxena, D., Bais, M.V. (2017). TGF-β1- and CCN2-Stimulated Sirius Red Assay for Collagen Accumulation in Cultured Cells. In: Takigawa, M. (eds) CCN Proteins. Methods in Molecular Biology, vol 1489. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6430-7_39

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  • DOI: https://doi.org/10.1007/978-1-4939-6430-7_39

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6428-4

  • Online ISBN: 978-1-4939-6430-7

  • eBook Packages: Springer Protocols

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