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.

Assay Type: Skin
Tissue: Skin Disease Biopsies
Target: Study Dependant
Control Compound: Betamethasone
Study Type: Ex vivo cultures
Functional Endpoint: Inflammation

Testing Information

Introduction

The specific results that will be provided are on the anti-inflammatory effect of test articles across atopic dermatitis biopsies.

Turnaround Time

For a standard experiment with n=5–10, tissue procurement is estimated to be 8-10 weeks. These timelines are a guide based on standard tissue criteria; any client-imposed alterations to the criteria may alter these estimates.

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.

Suggested Testing

We suggest test conditions are assessed in singlet across 5–10 donors

The set up for a typical full-thickbness human skin tissue explant experiment


Study Outline

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)

Exclusion Criteria

No specific exclusion criteria are in place other than to reject macroscopically diseased/necrotic tissue.

Analysis

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.


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