Improved skin substitutes and uses thereof

Details Improved skin substitutes and uses thereof Improved skin substitutes and uses thereof

C - Chemistry, Metallurgy

12

N

C12N 5/10 (2006.01) C12N 5/071 (2010.01) A61K 35/36 (2006.01) C12N 15/63 (2006.01) C12N 15/85 (2006.01) C12N 15/12 (2006.01)

Patent

CA 2439899

The present invention relates to in vitro cultured skin substitutes, and in particular to in vitro cultured skin substitutes that have improved barrier function. In some embodiments, improved barrier function is a result of improved culture conditions, while in other embodiments, improved barrier function results from genetic modification of keratinocyts. Improved culture conditions to improve barrier function include organotypic culture in the presence of linoleic acid and/or linoleic acid at about 75% humidity. Suitable genetic modifications for improving barrier function includes transfection with a DNA construct capable of expressing GKLF. The present invention further provides improved methods of using such skin substitutes in the screening of compounds for irritancy activity, as well for identifying novel irritant responsive genes.

L'invention porte sur des substituts de peau cultivés in vitro, et plus particulièrement sur des substituts de peau cultivés in vitro qui ont amélioré leur fonction barrière. Dans certains modes de réalisation, la fonction barrière améliorée résulte de conditions de culture améliorées alors que dans d'autres modes de réalisation, la fonction barrière améliorée résulte de modifications génétiques des kératinocytes. Les conditions de culture améliorées améliorant la fonction barrière comprennent la culture organotypique en présence d'acide linoléique et/ou d'acide linoléique présentant environ 75 % d'humidité. Des modifications génétiques permettant d'améliorer la fonction barrière comprennent la transfection avec la construction d'ADN capable d'exprimer GKLF. L'invention concerne aussi des procédés améliorés d'utilisation de tels substituts de peau dans le criblage de composés pour l'activité d'irritation ainsi que pour l'identification de nouveaux gènes réceptifs à l'irritation.

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