Assaying proliferation and differentiation capacity of stem cells using disaggregated adult mouse epidermis
Abstract
In this protocol, we describe how to isolate keratinocytes from adult mouse epidermis, fractionate them into different sub-populations on the basis of cell surface markers and examine their function in an in vivo skin reconstitution assay with disaggregated neonatal dermal cells. We also describe how the isolated keratinocytes can be subjected to clonal analysis in vitro and in vivo and how to enrich for hair follicle-inducing dermal papilla cells in the dermal preparation. Using these approaches, it is possible to compare the capacity of different populations of adult epidermal stem cells to proliferate and to generate progeny that differentiate along the different epidermal lineages. Isolating, fractionating and grafting cells for the skin reconstitution assay is normally spread over 2 d. Clonal growth in culture is assessed after 14 d, while evaluation of the grafts is carried out after 4-5 weeks.
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Rapamycin increases survival and repopulating capacity of epithelial progenitors, protecting from radiation-induced loss of this tissue regenerative cell population.
“these findings indicate that rapamycin increases the survival and repopulating capacity of epithelial progenitors, hence protecting from radiation-induced loss of this tissue regenerative cell population.”
From mTOR inhibition prevents epithelial stem cell senescence and protects from radiation-induced mucositis (2012) · cites this paper as Jensen et al., 2010
Each large colony is expected to grow from a single surviving self-renewing epithelial stem cell.
“Since each large colony is expected to grow from a single surviving self-renewing epithelial stem cell (Jensen et al., 2010)”
From mTOR inhibition prevents epithelial stem cell senescence and protects from radiation-induced mucositis (2012) · cites this paper as Jensen et al., 2010
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References 39 · Cited by 178
- Authors
- Jensen, K. B., Driskell, R. R., & Watt, F. M.
- Affiliations
- Wellcome/MRC Cambridge Stem Cell Institute (GB)
- Corresponding author
- Watt, Fiona M
- Venue
- Nature Protocols
- Published
- 2010-04-22
- Volume
- 5
- Issue
- 5
- Pages
- 898-911
- ISSN
- 1750-2799 (ISSN-L)
- 1754-2189
- OpenAlex
- W1969737028
- Cited by
- 178
- References
- 39
- Citations by year
- 2026: 4 · 2025: 5 · 2024: 6 · 2023: 14 · 2022: 12 · 2021: 12 · 2020: 12 · 2019: 13 · 2018: 14 · 2017: 15 · 2016: 11 · 2015: 10 · 2014: 12 · 2013: 16 · 2012: 13
- FWCI
- 16.57
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Protocols (publishedVersion)
- PubMed (publishedVersion)
- Research Portal (King's College London) (submittedVersion)
- Funders
- Wellcome Trust
- Cancer Research UK
- Topics
- Health Sciences › Medicine › Urology › Hair Growth and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Skin and Cellular Biology Research
- Health Sciences › Medicine › Hematology › Hematopoietic Stem Cell Transplantation
- Keywords
- epidermal stem cells, keratinocytes, dermal papilla cells, hair follicle induction, cell surface markers, subcellular fractionation, clonal analysis, cell proliferation, cell differentiation, mouse epidermis
- MeSH terms
- Epidermal Cells, Animals, Cells, Cultured, Flow Cytometry, Mice, Adult Stem Cells
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