Isolation of Stem Cells and Progenitors from Mouse Epidermis

Lana Kostic1, Egor Sedov1, Despina Soteriou1, Yahav Yosefzon1, Yaron Fuchs1

1 Laboratory of Stem Cell Biology and Regenerative Medicine, Department of Biology and Lokey Interdisciplinary Center for Life Sciences and Engineering, Technion Israel Institute of Technology, Haifa
Publication Name:  Current Protocols in Stem Cell Biology
Unit Number:  Unit 1C.20
DOI:  10.1002/cpsc.26
Online Posting Date:  May, 2017
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The epidermis consists of several distinct compartments including the interfollicular epidermis (IFE), sweat glands, sebaceous glands (SGs), and the hair follicle (HF). While the IFE and SGs are in a constant state of self‐renewal, the HF cycles between phases of growth, destruction, and rest. The hair follicle stem cells (HFSCs) that fuel this perpetual cycle have been well described and are located in a niche termed the bulge. These bulge SCs express markers such as CD34 and Keratin 15 (K15), enabling the isolation of these cells. Here, we describe a powerful method for isolating HFSCs and epidermal progenitors from mouse skin utilizing fluorescence activated cell‐sorting (FACS). Upon isolation, cells can be expanded and utilized in various in vivo and in vitro models aimed at studying the function of these unique cells. © 2017 by John Wiley & Sons, Inc.

Keywords: cell sorting; epidermis; FACS; hair follicle; progenitors; skin; stem cells

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Table of Contents

  • Basic Protocol 1: Isolation of Hair Follicles, Stem Cells, and Keratinocytes from Adult Mouse Epidermis
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
  • Tables
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Basic Protocol 1: Isolation of Hair Follicles, Stem Cells, and Keratinocytes from Adult Mouse Epidermis

  • Isoflurane
  • Mice (50 to 80 days old; C57BL/6)
  • 70% ethanol
  • PBS without Ca2+ and Mg2+
  • 0.25% trypsin/EDTA
  • Staining buffer (see recipe)
  • DAPI
  • Integrin β1 antibody
  • Integrin α6 antibody
  • Sca‐1 antibody
  • CD34 antibody
  • Chelated fetal bovine serum (see recipe)
  • Anesthesia machine with induction box
  • CO 2 chamber (for euthanasia)
  • Electric shaver
  • Dissecting pad
  • Scissors
  • Pins
  • Blunt scalpel
  • Dissection forceps (flat and curved)
  • Culture dishes
  • Electric pipet filler
  • 50‐ml Falcon tubes
  • 70‐μm cell strainers
  • 40‐μm cell strainers
  • Centrifuge
  • FACS tubes
  • FACS tubes with cell strainer caps
  • Aluminum foil
  • Additional reagents and equipment for euthanizing mice (Donovan & Brown, )
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Literature Cited

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