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Development of in-vitro systems using human immortalised cell lines for testing skin irritancy


The testing of cosmetological (and pharmaceutical) preparations will have to be performed in a near future by procedures alternative to animal experiments to comply with the Sixth Amendment (93-35-EEC) to the EC Cosmetic Directive (976- 768-EEC). In vitro recombined skin using human cells represents an elegant procedure for this purpose.
The presently available models suffer imperfections that will be addressed in the present project to aim at the preparation of an in-vitro test for skin irritancy. It will display meaningful endpoints, should prove reproducible by the use of chemically defined culture products and immortalised cell lines and be easy-to-use by introducing adequate markers of the irritant reaction. The programme will be a shared activity of six academic research units representing a large part of the scientific european potential in the domain and associated industrial laboratories experts in the use of reconstructed skin for irritant testing.
The project will be developed in four partially overlapping steps addressing to defined problems. Step 1- improvement of the quality of the stratum corneum in-vitro, introduction of endothelial cells in the dermal compartment and determination of in-vivo related endpoints of irritancy. Step 2 - replacement of the human cell strains by immortalised human cell lines and determination of the conditions required to obtain near physiological functions and the above determined selected end-points of the irritant reaction. Step 3 introduction of the markers of the irritant reaction by infection or transfection of constructions made of a reporter gene driven by the promoter of a protein participating in the process in the different lines of cells (keratinocytes, fibroblasts and endothelial cells).
Step 4 - preparation of a model suitable for prevalidation testing in collaboration with industrial partners.
The benefits of the project is scientific by the european collaboration required to address to the specific know-how of the different types of cells included in the model, economical by helping the industry to comply to the EEC directives and ethical by a contribution to the faster suppression of testing in animals by an alternative procedure.

Funding Scheme

CSC - Cost-sharing contracts


Université de Liège

4000 Liège

Participants (5)

Institut National de la Santé et de la Recherche Médicale
Avenue Claude Vellefaux
75475 Paris
Istituto di Ricerche Farmacologiche Mario Negri
Via Eritrea 62
20157 Milano
Rijksuniversiteit Leiden
2300 RC Leiden
Stiftung Deutsches Krebsforschungszentrum
280,Im Neuenheimer Feld
69120 Heidelberg
Universität Köln
9,Joseph Stelzmann Straße
50931 Köln