Abstract
Culturing keratinocytes to form coherent epithelial tissue sheets has improved the treatment of extensively burned patients. Keratinocyte culture is also used to investigate various cellular and molecular mechanisms involved in different skin pathologies. To preserve stem cells during epithelial cell culture, reliable methods and conditions are of the utmost importance. Properly cultured keratinocytes will exhibit a consistent cuboid morphology and can proliferate for many passages. This chapter details materials needed and methods for all aspects of efficient keratinocyte culture for clinical applications, namely tissue sampling and transportation, isolation, routine culture, subculture, and cryopreservation.
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Acknowledgments
The authors would like to thank the current and former members of the LOEX who have contributed to develop these protocols. This work was supported by the Canadian Institutes for Health Research (CIHR) grants MOP-12087 and FDN-143213 (L.G.), the Fondation des Pompiers du Québec pour les Grands Brûlés (FPQGB), the Fonds de Recherche du Québec-Santé (FRQS), and the Réseau de thérapie cellulaire et tissulaire (ThéCell) du FRQS. L.G. is the holder of a Tier 1 Canadian Research Chair from CIHR on Stem Cells and Tissue Engineering, and a Research Chair from the Fondation de l’Université Laval on Tissue-Engineered Organs and Translational Medicine.
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Cortez Ghio, S., Le-Bel, G., Lavoie, A., Larouche, D., Germain, L. (2019). Isolation and Culture of Human Keratinocytes. In: Böttcher-Haberzeth, S., Biedermann, T. (eds) Skin Tissue Engineering. Methods in Molecular Biology, vol 1993. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9473-1_1
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DOI: https://doi.org/10.1007/978-1-4939-9473-1_1
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