Effects of undigested protein-rich ingredients on polarised small intestinal organoid monolayers

Abstract

Here, we describe the use of monolayers of intestinal epithelial cells derived from intestinal organoids and transcriptomics to investigate the direct effects of dietary protein sources on epithelial function. Mechanically dissociated 3D organoids of mouse duodenum were used to generate a polarized epithelium containing all cell types found in the tissue of origin. The organoid-derived cell monolayers were exposed to 4% (w/v) of 'undigested (non-hydrolysed)-soluble' fraction of protein sources used as feed ingredients [soybean meal (SBM) and casein], or alternative protein sources (spray dried plasma protein, and yellow meal worm), or controls for 6 h prior to RNA isolation and transcriptomics. All protein sources altered expression of unique biological processes in the epithelial cells. Exposure of intestinal organoids to SBM downregulated expression of retinol and retinoid metabolic processes as well as cholesterol and lipid biosynthetic pathways, consistent with the reported hypotriglyceridaemic effect of soy protein in vivo. These findings support the use of intestinal organoids as models to evaluate complex interactions between dietary ingredients and the intestinal epithelium and highlights some unique host effects of alternative protein sources in animal feed and potentially human food.

Congruence of Transcription Programs in Adult Stem Cell-Derived Jejunum Organoids and Original Tissue During Long-Term Culture

“…The spatial nature of 3-dimensional organoids prevents easy access to the apical surface, but can be overcome by injection or monolayer formation. We have recently shown that apical stimulation of small intestinal organoids with various protein rich ingredients elicit specific metabolic responses, like altered lipid metabolism (Kar et al, 2020). In that study, predicted putative pathways were verified using protein-detection assays, showing the ability to measure gene-predicted responses in organoids.…”

Organoids: a promising new in vitro platform in livestock and veterinary research

“…Furthermore, the niche factors and MG contribute substantially to the high costs of organoid culture. For the culture of mouse intestinal organoids it was shown that epidermal growth factor can be replaced by lysophosphatidic acid [ 112 ]. Similarly, for the culture of chicken intestinal organoids, the expensive agonists R-spondin 1 and Noggin may be replaced by prostaglandin E2 [ 92 ].…”

“…In the area of animal nutrition, intestinal organoids may be used to study the gastro-intestinal epithelial response to feed ingredients (as has been done with mouse organoids [ 112 ]). Performing such studies could aid nutritionist to determine “non-strict-nutritional” properties e.g.…”

“…To enable access to the apical membrane of epithelial organoids, which is necessary to study e.g. interaction with microbes or transport of nutrients, 2D organoid models have been developed [41][42][43][110][111][112]. It is necessary for farm animals that such 2D models are developed and validated.…”

502. Organoids as a research tool in animal breeding and nutrition

“…These structures, called organoids, contain multiple cell types, resemble the in vivo tissue, and recapitulate key function(s) of the organ. It has been demonstrated that organoids can be dissociated and seeded onto Matrigel-or collagen-coated Transwell membranes to form two-dimensional (2D) epithelial monolayers (Foulke-Abel et al, 2014;Van der Hee et al, 2020). These 2D cultures allow the study of interactions with apically applied exogenous stimuli, such as pathogens or compounds, i.e.…”