Skin Biopsies from Atopic Dermatitis Donors
Drug Discovery Assay – reference number: B118
This experiment assesses whether test articles cause a reduction in inflammatory cytokine release in atopic dermatitis skin biopsies, with Betamethasone as a reference compound. It uses clinical skin biopsies obtained from atopic dermatitis donors to explore test article effect on the expression on inflammatory cytokines via ELISA and/or rtPCR.
|Tissue:||Skin Disease Biopsies|
|Study Type:||Ex vivo cultures|
The specific results that will be provided are on the anti-inflammatory effect of test articles across atopic dermatitis biopsies.
Test Article Requirements
Test article(s) to be provided by the Sponsor in storable aliquots at required test concentrations with information on diluent vehicle used. Stock solutions are prepared in deionized water unless otherwise requested. Sponsor to provide sufficient test article to run the entire study.
We suggest test conditions are assessed in singlet across 5–10 donors
Diagram showing a typical setup for our skin disease explant assays
Rationale and Experimental Design
Full-thickness skin biopsies are obtained from our clinical networks. Biopsies are taken at a size of 3mm2 and then transferred into fortified media, leaving the epidermis exposed to air. Biopsies are incubated in optimum conditions for a maximum of 24 hrs, then cultured in the presence of the test article for a further 24 hrs. Tissue media is collected for phenotype analysis by ELISA and/or the biopsy set is collected for rtPCR.
An example of the conditions assessed for 1 test article is detailed below (it is recommended a minimum of 5–10 donors should be used for each condition):
- Control (diseased biopsy)
- Test article (treated, disease biopsy)
- Control 2 (non-diseased biopsy)
No specific exclusion criteria are in place other than to reject macroscopically diseased/necrotic tissue.
Cytokine analysis: Supernatant samples can be analyzed for specific analytes of your choice by a multiplex ELISA platform. Each analyte will be quantified by interpolation against a standard curve generated on the same 96 well analysis plate.
Other forms of end point analysis are available such as gene expression or immunohistochemistry.