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A Simplified and Efficient Method to Isolate Primary Human Keratinocytes from Adult Skin Tissue
Published on: August 25, 2018
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A Newly Developed Chemically Defined Serum-Free Medium Suitable for Human Primary Keratinocyte Culture and Tissue
Sergio Cortez Ghio1,2,3, Martin A Barbier1,2,3, Emilie J Doucet1,2,3
1The Tissue Engineering Laboratory (LOEX), a Université Laval's Research Center, QC G1V 0A6, Canada.
International Journal of Molecular Sciences
|February 11, 2023
Summary
A new serum-free keratinocyte culture medium, Surge SFM, enables efficient expansion of primary human keratinocytes and production of functional skin substitutes for research and clinical use.
Area of Science:
- Cell Biology
- Tissue Engineering
- Dermatology
Background:
- Conventional serum-containing media limit massive expansion and stratification of human keratinocytes.
- Existing defined, serum-free media are insufficient for large-scale keratinocyte expansion and skin substitute generation.
Purpose of the Study:
- To develop and evaluate a novel chemically defined, serum-free medium (Surge SFM) for massive expansion of human primary keratinocytes.
- To assess Surge SFM's capability in generating functional skin substitutes for potential clinical applications.
Main Methods:
- Development of Surge SFM for feeder-layer-supported keratinocyte culture.
- Evaluation of keratinocyte proliferation, stem cell retention (keratin-19+), and gene expression.
- Production and histological assessment of bilayered skin substitutes.
- Grafting of skin substitutes onto athymic mice for in vivo evaluation.
Main Results:
- Surge SFM efficiently supports massive expansion of both freshly isolated and cryopreserved primary human keratinocytes over multiple passages.
- Keratin-19-positive epithelial stem cells are retained during serial passaging in Surge SFM.
- Transcriptomic analysis shows gene expression similar to serum-containing media.
- Surge SFM facilitates the generation of bilayered skin substitutes with histology comparable to serum-containing media.
- Grafted skin substitutes demonstrated persistence for up to six months in vivo.
Conclusions:
- The developed Surge SFM is a promising chemically defined, serum-free medium for large-scale human keratinocyte expansion.
- Surge SFM enables the production of functional skin substitutes suitable for research and clinical translation.
- This advancement offers a viable alternative to serum-containing media in keratinocyte culture and tissue engineering.
