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Isolation of Mouse Epidermal Keratinocytes and Their In Vitro Clonogenic Culture
Published on: August 10, 2019
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Global gene expression and comparison between multiple populations in the mouse epidermis.
Anders Patrik Gunnarsson1, Rikke Christensen2, Jian Li3
1Department of Clinical Genetics, Aarhus University Hospital, 8200 Aarhus, Denmark; Institute of Clinical Medicine, Aarhus University, 8000 Aarhus, Denmark.
Stem Cell Research
|July 25, 2016
Summary
This study identifies novel stem cell populations in mouse hair follicles using surface markers and transcriptome profiling. Cultured cells lose original characteristics, revealing discrepancies between isolated and cultured cells.
Area of Science:
- Stem cell biology
- Dermatology
- Molecular biology
Background:
- Mouse hair follicles harbor stem cell populations identified by surface markers.
- Previous studies have characterized some of these populations.
Purpose of the Study:
- To determine if a multicolor panel of CD34, Sca-1, Integrin-α6, and Plet-1 can sufficiently separate known hair follicle stem cell populations.
- To characterize these populations using global transcriptome profiling.
- To analyze transcriptome profiles of cultured populations.
Main Methods:
- Utilized flow cytometry with a multicolor panel of four surface markers (CD34, Sca-1, Integrin-α6, Plet-1).
- Performed global transcriptome profiling on isolated cell populations.
- Conducted transcriptome analysis on selected populations after two weeks of culturing.
Main Results:
- Identified eight distinct cell populations, including two previously undescribed subsets: one expressing only Integrin-α6 and another expressing all markers except CD34.
- Both novel subsets demonstrated high clonogenic potential.
- Transcriptome profiling revealed Hspa2 expression in a population negative for all tested markers, identified as inner root sheath keratinocytes via immunostaining.
- All cultured cell populations lost their original characteristics and could not be distinguished by gene expression profiles.
Conclusions:
- Multicolor flow cytometry panels can identify additional stem cell subsets within the epidermis.
- Significant discrepancies exist in gene expression between directly isolated cells and tissue-cultured cells.
- Further research is needed to understand the behavior and characteristics of hair follicle stem cell populations in vitro.
