|Tissue:||Human GI Mucosal Explants (IBD)|
|Target:||p38 MAP Kinase|
|Study Type:||Ex vivo Cultures|
This experiment assesses whether test articles cause a reduction in TNF-alpha release with BIRBR796 as a reference compound.
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).
The specific results that will be provided are the effects of test articles on the levels of TNF-alpha release in human IBD tissue.
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.
A minimum of triplicate per condition with a maximum of 18 biopsies in total.
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):
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.