Preparation of single-cell suspensions for cytofluorimetric analysis from different mouse skin regions

Achille Broggi, Clara Cigni, Ivan Zanoni, Francesca Granucci

Research output: Contribution to journalArticlepeer-review

Abstract

The skin is a barrier organ that interacts with the external environment. Being continuously exposed to potential microbial invasion, the dermis and epidermis home a variety of immune cells in both homeostatic and inflammatory conditions. Tools to obtain skin cell release for cytofluorimetric analyses are, therefore, very useful in order to study the complex network of immune cells residing in the skin and their response to microbial stimuli. Here, we describe an efficient methodology for the digestion of mouse skin to rapidly and efficiently obtain single-cell suspensions. This protocol allows maintenance of maximum cell viability without compromising surface antigen expression. We also describe how to take and digest skin samples from different anatomical locations, such as the ear, trunk, tail, and footpad. The obtained suspensions are then stained and analyzed by flow cytometry to discriminate between different leukocyte populations.

Original languageEnglish
Article numbere52589
JournalJournal of visualized experiments : JoVE
Volume2016
Issue number110
DOIs
Publication statusPublished - Apr 1 2016

Keywords

  • Collagenase
  • Dermis
  • Flow cytometry
  • Immune system
  • Issue 110
  • Leukocytes
  • Medicine
  • Single-cell preparation
  • Skin

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

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