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Drug Discovery Assay Services Assay: IBD culture study Crohn's or ulcerative colitis (UC)

Code: B106
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Tissue:Human GI Mucosal Explants (IBD)
Target:p38 MAP Kinase
Control Compound:BIRB796
Study Type:Ex vivo Cultures
Functional Endpoint:Inflammation

Assay Description

This experiment assesses whether test articles cause a reduction in TNF-alpha release with BIRBR796 as a reference compound.

Example assay graph

Image Legend

Figure 1: Graph demonstrating a reduction in TNF-alpha release in stimulated full thickness mucosal biopsies from Crohn's and UC donors (n = 16). The mean reduction = 56.49 % ± 5.76 (mean ± S.E.M).

Testing Information

Introduction

The specific results that will be provided are the effects of test articles on the levels of TNF-alpha release in human IBD tissue.

Turnaround Time

6 weeks from sample receipt for n = 3 donors.

Timelines are a guide based on REPROCELL's standard tissue criteria, any client-imposed deviations to the standard tissue criteria can alter the project timelines.

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 distilled water unless otherwise requested. Sponsor to provide sufficient test article to run the entire study.

Suggested Testing

A minimum of triplicate per condition with a maximum of 18 biopsies in total.

Study Outline

Rationale and Experimental Design

Full thickness gut mucosal biopsies are dissected from fresh Crohn's or UC tissue samples. Biopsies are taken at a size of 5mm2 and transferred to a plate containing specially fortified media before being incubated in optimum conditions for a maximum of 24 hrs. Biopsies are cultured in the presence of an inflammatory stimulant to aid in the normalisation of cytokine levels.

An example of the conditions assessed for 3 test articles are detailed below (it is recommended a minimum of triplicates should be used for each condition):

  • Non-stimulated control
  • Stimulated control
  • Test article 1
  • Test article 2
  • Test article 3
  • Positive control

Exclusion Criteria

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

End point analysis

Cytokine analysis: Supernatant samples can be analysed 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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