High-frequency transfection of cultured human epidermal basal cells that differentiate to form a multilayered tissue.
Primary cultures of human keratinocytes form a multilayered tissue. By incubating the tissue cultures in Ca2(+)-free medium the differentiated cell layers can be stripped off leaving a basal cell monolayer. We have developed a method for high-frequency transfection of these epidermal basal cells with genes inserted into Epstein-Barr virus-based expression vectors. Using the Escherichia coli lac z gene as a marker gene, the transient and long-term expression and the fate of the transfected cells were studied. During regeneration of the multilayered tissue most of the transfected basal cells enlarge and undergo differentiation, but a minor population remains as basal cells. Incubation with the tumor promotor 12-O-tetradecanoylphorbol-13-acetate results in an increase in the proportion of transfected keratinocytes that are small, suggesting a relative expansion of the immature cell pool.
Jensen
T G
TG
Institute of Human Genetics, University of Aarhus, Denmark.
Jensen
P K
PK
Nørgård
J O
JO
Bolund
L
L
D000328:Adult; D002454:Cell Differentiation; D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods; D000078404:Epidermal Cells; D005814:Genes, Viral; D005822:Genetic Vectors; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000187:drug effects; D007700:Kinetics; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection
D013755:Tetradecanoylphorbol Acetate