Recapitulation of the accessible interface of biopsy-derived canine intestinal organoids to study epithelial-luminal interactions

Abstract

Recent advances in canine intestinal organoids have expanded the option for building a better in vitro model to investigate translational science of intestinal physiology and pathology between humans and animals. However, the three-dimensional geometry and the enclosed lumen of canine intestinal organoids considerably hinder the access to the apical side of epithelium for investigating the nutrient and drug absorption, host-microbiome crosstalk, and pharmaceutical toxicity testing. Thus, the creation of a polarized epithelial interface accessible from apical or basolateral side is critical. Here, we demonstrated the generation of an intestinal epithelial monolayer using canine biopsy-derived colonic organoids (colonoids). We optimized the culture condition to form an intact monolayer of the canine colonic epithelium on a nanoporous membrane insert using the canine colonoids over 14 days. Transmission and scanning electron microscopy revealed a physiological brush border interface covered by the microvilli with glycocalyx, as well as the presence of mucin granules, tight junctions, and desmosomes. The population of stem cells as well as differentiated lineagedependent epithelial cells were verified by immunofluorescence staining and RNA in situ hybridization. The polarized expression of P-glycoprotein efflux pump was confirmed at the apical membrane. Also, the epithelial monolayer formed tight-and adherence-junctional barrier within 4 days, where the transepithelial electrical resistance and apparent permeability were inversely correlated. Hence, we verified the stable creation, maintenance, differentiation, and physiological function of a canine intestinal epithelial barrier, which can be useful for pharmaceutical and biomedical researches.

Three-Dimensional Regeneration of Patient-Derived Intestinal Organoid Epithelium in a Physiodynamic Mucosal Interface-on-a-Chip

“…Furthermore, the enclosed lumen in an organoid body considerably hampers access to the lumen, by which the co-culture with gut microbiome or the simulation of drug administration is extremely challenging. Thus, this limitation led to the innovation of an “opened” version of an organoid, where the recreation of a mucosal tissue interface of intestinal organoids has been emerged [ 6 , 48 , 49 ]. On top of the previous reports [ 20 , 50 ], we propose an advanced 3D convoluted microdevice that can manipulate multiaxial deformations and independently controlled fluid hydrodynamics in the mucosal interface.…”

Treatment With Hydrolyzed Diet Supplemented With Prebiotics and Glycosaminoglycans Alters Lipid Metabolism in Canine Inflammatory Bowel Disease

“…The present study showed a positive effect of IBD treatment using PRBs and GAGs in addition to the hydrolyzed diet alone. Mechanistic effects of glycerophospholipids or TGs could be tested in our recently established intestinal cell models to better recapitulate canine intestine (54,55).…”

Emerging Roles of Urine-Derived Components for the Management of Bladder Cancer: One Man’s Trash is Another Man’s Treasure

“…Patient-derived xenografts (PDX) is an approach whereby patient tumor fragments are implanted into immunocompromised mice to generate tumors that recapitulate genomic and phenotypical features of patient's original tumor 30,31 . PDX have value in both better understanding tumor biology and evaluating the efficacy of FDA-approved anticancer therapies or novel targeted treatments 37 . Although PDX models present an exciting opportunity for improving predictive value of preclinical studies, there are several hurdles to their translation into the clinic.…”

“…In addition, xenografted urine-derived organoids were able to cause tumorigenesis in immunodeficient mice demonstrating their ability to maintain oncogenic properties ex vivo. Characterization of organoids can be done through an array of cellular and molecular techniques including immunohistochemistry, RNA-Seq, proteomics, and others 37,38,39 . Leveraging our expertise in the culture and maintenance of canine organoids, our consortium has successfully cultured MIBC organoids from urinary samples of 6/6 human patients ( Fig.…”

“…Leveraging our expertise in the culture and maintenance of canine organoids, our consortium has successfully cultured MIBC organoids from urinary samples of 6/6 human patients ( Fig. 1) 37,38,39 . Immunohistochemistry staining for Gata 3, p63 and Pax 8 (Gata 3+, p63+, Pax 8-) confirmed the urothelial origin of 3D organoids ( Fig.…”