In vitro culture conditions to study keratinocyte differentiation using the HaCaT cell line
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In vitro models to study the process of keratinocyte differentiation have been hindered by the stringent culture requirements and limitations imposed by the inherent properties of the cells. Primary keratinocytes only have a finite life span, while transformed cell lines exhibit many phenotypic features not found in normal cells. The spontaneously immortalized HaCaT cell line has been a widely employed keratinocyte model due to its ease of propagation and near normal phenotype, but protocols for differentiation and gene delivery into HaCaT cells vary widely in the literature. Here we report culture conditions for maintaining HaCaT cells in a basal-like state, for efficient differentiation of these cells, and for delivery of transgenes by transfection or adenoviral infection. This technological report will provide guidance to a large audience of scientists interested in investigating mechanisms of differentiation and skin morphogenesis.
KeywordsHaCaT Differentiation Keratinocyte Transfection Infection Calcium
We greatly thank Dr. Bokoch (La Jolla, California) for providing us with HaCaT, a naturally immortalized, human keratinocyte cell line. We also acknowledge Dr. Davis (Texas A&M Health Science Center, Department of Pathology and Laboratory Medicine) for supplying the Ad5-GFP vector. The work was supported by a grant from the National Cancer Institute (R01 CA089298)to V.G.W. and a Life Sciences Training fellowship to A.F.D.
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