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Isolation of Live Fibroblasts by Fluorescence-Activated Cell Sorting

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Fibrosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1627))

Abstract

Flow cytometry is a powerful tool in cell biology in that it allows real-time characterization of cellular phenotypes. Additionally, through the process of fluorescence-activated cell sorting (FACS), living cells can be isolated for future in vitro experiments, including single cell analysis. Here we describe the isolation of live fibroblasts from the dermis of the skin in mice using FACS. This method circumvents the need for ex vivo expansion in cell culture, which can alter phenotypic and functional characteristics of cells.

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References

  1. Walmsley GG, Rinkevich Y, Hu MS et al (2015) Live fibroblast harvest reveals surface marker shift in vitro. Tissue Eng Part C Methods 21(3):314–321. doi:10.1089/ten.TEC.2014.0118

  2. Walmsley GG, Maan ZN, Hu MS et al (2016) Murine dermal fibroblast isolation by FACS. J Vis Exp 107. doi:10.3791/53430

  3. Rinkevich Y, Walmsley GG, Hu MS et al (2015) Skin fibrosis. Identification and isolation of a dermal lineage with intrinsic fibrogenic potential. Science 348(6232):aaa2151. doi:10.1126/science.aaa2151

    Article  PubMed  PubMed Central  Google Scholar 

  4. Crissman HA, Mullaney PF, Steinkamp JA (1975) Methods and applications of flow systems for analysis and sorting of mammalian cells. Methods Cell Biol 9(0):179–246

    Article  CAS  PubMed  Google Scholar 

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Author information

Authors and Affiliations

  1. Hagey Laboratory for Pediatric Regenerative Medicine, Division of Plastic and Reconstructive Surgery, Department of Surgery, School of Medicine, Stanford University, Stanford, CA, USA

    Tripp Leavitt, Michael S. Hu & Michael T. Longaker

  2. Boston University School of Medicine, Boston, MA, USA

    Tripp Leavitt

  3. Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, Stanford, CA, USA

    Michael S. Hu & Michael T. Longaker

  4. Department of Surgery, John A. Burns School of Medicine, University of Hawai’i, Honolulu, HI, USA

    Michael S. Hu

Authors
  1. Tripp Leavitt
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  2. Michael S. Hu
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  3. Michael T. Longaker
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Corresponding author

Correspondence to Michael T. Longaker .

Editor information

Editors and Affiliations

  1. Department of Dermatology, University of Michigan Medical School, Ann Arbor, Michigan, USA

    Laure Rittié

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Leavitt, T., Hu, M.S., Longaker, M.T. (2017). Isolation of Live Fibroblasts by Fluorescence-Activated Cell Sorting. In: Rittié, L. (eds) Fibrosis. Methods in Molecular Biology, vol 1627. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7113-8_13

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  • DOI: https://doi.org/10.1007/978-1-4939-7113-8_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7112-1

  • Online ISBN: 978-1-4939-7113-8

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