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Fibrosis pp 205-212 | Cite as

Isolation of Live Fibroblasts by Fluorescence-Activated Cell Sorting

  • Tripp Leavitt
  • Michael S. Hu
  • Michael T. Longaker
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

Fibroblasts Skin Flow cytometry Fluorescence-activated cell sorting Dermis Dermal tissue harvest Live cell isolation 

References

  1. 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. 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. 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 CrossRefPubMedPubMedCentralGoogle Scholar
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    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):179–246CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Tripp Leavitt
    • 1
    • 2
  • Michael S. Hu
    • 1
    • 3
    • 4
  • Michael T. Longaker
    • 1
    • 3
  1. 1.Hagey Laboratory for Pediatric Regenerative Medicine, Division of Plastic and Reconstructive Surgery, Department of SurgerySchool of Medicine, Stanford UniversityStanfordUSA
  2. 2.Boston University School of MedicineBostonUSA
  3. 3.Institute for Stem Cell Biology and Regenerative MedicineStanford UniversityStanfordUSA
  4. 4.Department of SurgeryJohn A. Burns School of Medicine, University of Hawai’iHonoluluUSA

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