Institut for Biomedicin

Lars Bolund

High-frequency transfection of cultured human epidermal basal cells that differentiate to form a multilayered tissue

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  • Plantepatologi og Entomologi
  • Institut for Antropologi, Arkæologi og Lingvistik
  • Institut for Human Genetik

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.

OriginalsprogEngelsk
TidsskriftExperimental Cell Research
Vol/bind189
Nummer2
Sider (fra-til)163-8
Antal sider6
ISSN0014-4827
StatusUdgivet - aug. 1990

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