A simple reconstructed human epidermis: preparation of the culture model and utilization in in vitro studies

Yves Poumay, Frederic Dupont, Sarah Marcoux, Michèle Smekens, Michel Herin, Alain Coquette

Résultats de recherche: Contribution à un journal/une revueArticle

Résumé

The preparation of a reconstructed human epidermis is described with examples of its utilization in in vitro studies. The model was obtained by culturing normal human keratinocytes at high cell density for 14 days in serum-free and high calcium (1.5 mM) medium on an inert polycarbonate filter at the air-liquid interface. These stratified cultures showed histological features similar to those observed in vivo in the epidermis: a proliferating basal layer and differentiating spinous, granular, and cornified layers. Electron microscopy illustrated lamellar bodies, junctions and keratohyalin granules. Immunofluorescent localization of epidermal markers (keratins 14 and 10, involucrin and filaggrin) revealed typical differentiation. This in vitro reconstructed tissue was used in studies of toxic effects of chemicals. The modelled tissue showed progressive cytotoxicity of a skin irritant (benzalkonium chloride) and a sensitizer (dinitrochlorobenzene) as assessed by MTT assay. Moreover, differential release of interleukin-1α and interleukin-8 were measured after 20 h of incubation allowing the irritant to be distinguished from the sensitizer. Permeation studies indicated efficient barrier function of the reconstructed epidermis, as well as metabolizing properties towards hormones. This model can be custom-made and is potentially useful for studies involving keratinocytes in the epidermis, in basic science, dermatology or toxicology.
langue originaleAnglais
Pages (de - à)203-211
Nombre de pages9
journalArchives of Dermatological Research
Volume296
Les DOIs
Etat de la publicationPublié - 2004

Empreinte digitale

Examiner les sujets de recherche de « A simple reconstructed human epidermis: preparation of the culture model and utilization in in vitro studies ». Ensemble, ils forment une empreinte digitale unique.

Contient cette citation