Establishing a High Throughput Epidermal Spheroid Culture System to Model Keratinocyte Stem Cell Plasticity

Epithelial dysregulation is a node for a variety of human conditions and ailments, including chronic wounding, inflammation, and over 80% of all human cancers. As a lining tissue, the skin epithelium is often subject to injury and has evolutionarily adapted by acquiring the cellular plasticity neces...

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Published inJournal of visualized experiments no. 167
Main Authors Woappi, Yvon, Ezeka, Geraldine, Vercellino, Justin, Bloos, Sean M., Creek, Kim E., Pirisi, Lucia
Format Journal Article
LanguageEnglish
Published United States 30.01.2021
Subjects
Biomarkers - metabolism
Cell Culture Techniques - methods
Cell Plasticity
Cell Proliferation
Cell Separation
Cell Tracking
Cells, Cultured
Colony-Forming Units Assay
Epidermal Cells - cytology
epithelial cells
epithelium
Humans
inflammation
keratinocytes
Keratinocytes - cytology
Male
plasticity
Spheroids, Cellular - cytology
stem cells
Stem Cells - cytology
Transcription, Genetic
Online AccessGet full text
ISSN1940-087X
1940-087X
DOI10.3791/62182

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Summary:Epithelial dysregulation is a node for a variety of human conditions and ailments, including chronic wounding, inflammation, and over 80% of all human cancers. As a lining tissue, the skin epithelium is often subject to injury and has evolutionarily adapted by acquiring the cellular plasticity necessary to repair damaged tissue. Over the years, several efforts have been made to study epithelial plasticity using in vitro and ex vivo cell-based models. However, these efforts have been limited in their capacity to recapitulate the various phases of epithelial cell plasticity. We describe here a protocol for generating 3D epidermal spheroids and epidermal spheroid-derived cells from primary neonatal human keratinocytes. This protocol outlines the capacity of epidermal spheroid cultures to functionally model distinct stages of keratinocyte generative plasticity and demonstrates that epidermal spheroid re-plating can enrich heterogenous normal human keratinocytes (NHKc) cultures for integrinα6 /EGFR keratinocyte subpopulations with enhanced stem-like characteristics. Our report describes the development and maintenance of a high throughput system for the study of skin keratinocyte plasticity and epidermal regeneration.
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ISSN:1940-087X
1940-087X
DOI:10.3791/62182