Apical-Out Chicken Enteroids as an In-Vitro Model to Study Effects of an Alternative Growth Promoter on Indicators of Gut Health
Date
2023
Authors
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Publisher
University of Delaware
Abstract
Recent restrictions on antibiotic use in food animals has prompted the investigation of natural alternatives for promotion of growth efficiency and gut health. While in-vivo research has traditionally been used for investigation of such compounds, a shift toward in-vitro research is desirable for reduction of live animal involvement and associated costs. A novel in-vitro method, apical-out chicken enteroids, is characterized and evaluated here for its use in investigating changes in gene expression, gut barrier function, reactive oxygen species production, and apoptosis using RT-qPCR and functional fluorescence-based assays when challenged after pre-treatment with a potential antibiotic growth promoter alternative. Optimization of methods and results indicated various limitations with the apical-out chicken enteroids including inconsistent RNA yields and quality, high inter-well variation in enteroid size and number, and lack of replicability among fluorescence assay results. This served as valuable insight regarding improvements for the novel method of culture, but disallowed conclusive results to be drawn about the abilities of the potential antibiotic growth promoter alternative (AVT Weaning Oil). Further, the lack of assay replicability observed was hypothesized to be associated with inadequate methods of normalization for variable enteroid size and number, thus normalization to cumulative enteroid area opposed to enteroid counts was also assessed and deemed more desirable for future work with this chicken intestinal model.
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Keywords
Cell culture, Chicken enteroids, Gut health, Organoids, Intestinal model