Undergraduate Thesis Or Project
 

MatTek™ EpiIntestinal™ a 3D Small Intestine Epithelial Cell Culture Model For In Vitro Screening in Dietary Chemopreventive Agents

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https://ir.library.oregonstate.edu/concern/undergraduate_thesis_or_projects/cf95jk59v

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  • Testing the many potential chemoprevention agents from dietary sources in animal or human models is expensive and takes a long time. In vitro models can be useful for screening the right compounds to use in larger studies. We tested the MatTek™ EpiIntestinal™ small intestine epithelial 3D cell culture model for its use as a screening tool. Two experiments were conducted using 80 µM and 200 µM benzo[a]pyrene (BaP) in the first. BaP was used to establish the presence of Phase I and Phase II enzymes in this model. The second experiment tested chemo preventative agents 3,3’-diindolylmethane (DIM) (7.5 µM), sulforaphane (SFN) (7.5 µM), DIM+SFN at 7.5 µM each, and tryptophan (Trp) (1mM). Cells were treated for 48 hr. Transepithelial electrical resistance (TEER) was measured at 0 and 48 hours to assess barrier integrity. Lactate dehydrogenase (LDH) and gene regulation of selected metabolizing enzymes and barrier proteins were measured at 48 hours. BaP (200 µM) decreased TEER (p=0.0275) at 48 hr but no other treatment had an effect compared to vehicle controls. BaP (80 µM) increased LDH leakage at 48 hours (p≤0.01) compared to vehicle control. CYP1A1 and CYP1B1 were significantly upregulated by BaP in the first experiment (p≤0.0001) and by DIM, SFN+DIM, and Trp (p≤0.01) in the second experiment. The barrier protein Claudin-8 was upregulated by DIM+SFN (p≤0.05) and Trp (p≤0.001). Gap junction protein-β was up-regulated by SFN (p≤0.05), DIM+SFN (p≤0.05), and Trp (p≤0.01). This 3D intestinal epithelial model would be a useful in vitro screening tool for prioritizing chemopreventive agents for further research.
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