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Pig Taste Cell-Derived Organoids Synthesize Insulin

ABSTRACT: Previously we showed that taste receptor cells in situ in taste buds synthesize insulin. Here we describe a model of pig taste organoid culture in which we have promoted insulin expression by induction of quiescence. The cellular heterogeneity of the lingual epithelium is maintained in the organoids, and stem cell type and organoid architecture can be controlled through changes in media composition and/or use of static versus dynamic culture. Pig taste organoids were maintained long term and organoids cultured in low sheer stress dynamic exhibited an architecture and expression profile akin to the native tissue. Porcine taste organoids also contained insulin, and the insulin critical transcription factors MAFA and PAX4. These results provide a pig model of taste organoid culture that can be used universally and bring us closer to the use of the taste tissue as a new renewable source of beta cells

ORGANISM(S): Sus scrofa

PROVIDER: GSE252240 | GEO | 2024/01/31

REPOSITORIES: GEO

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Project description:In Rspondin-based three-dimensional cultures, Lgr5 stem cells from multiple organs form ever-expanding epithelial organoids that retain their tissue identity. Here we report the establishment of tumor organoid cultures from 20 consecutive colorectal carcinoma (CRC) patients. For most, organoids were also generated from adjacent normal tissue. Organoids closely resemble the original tumor. The spectrum of genetic changes within the 'living biobank' agrees well with previous large-scale mutational analyses of CRC. Gene expression analysis indicates that the major CRC molecular subtypes are represented. Tumor organoids are amenable to high-throughput drug screens allowing detection of gene-drug associations. As an example, a single organoid culture was exquisitely sensitive to Wnt secretion (porcupine) inhibitors and carried a mutation in the negative Wnt feedback regulator RNF43, rather than in APC. Organoid technology may fill the gap between cancer genetics and patient trials, complement cell line- and xenograft-based drug studies and allow personalized therapy design. Self-renewal of the intestinal epithelium is driven by Lgr5 stem cells located in crypts. We have recently developed a long-term culture system that maintains basic crypt physiology. Wnt signals are required for the maintenance of active crypt stem cells. Indeed, the Wnt agonist R-spondin1 induces dramatic crypt hyperplasia in vivo. R-spondin-1 is the ligand for Lgr5. Epidermal growth factor (EGF) signaling is associated with intestinal proliferation, while transgenic expression of Noggin induces a dramatic increase in crypt numbers. The combination of R-spondin-1, EGF, and Noggin in Matrigel sustains ever-expanding small intestinal organoids, which display all hallmarks of the original tissue in terms of architecture, cell type composition, and self-renewal dynamics. We adapted the culture condition for long-term expansion of human colonic epithelium and primary colonic adenocarcinoma, by adding nicotinamide, A83-01 (Alk inhibitor), Prostaglandin E2 and the p38 inhibitor SB202190. Of note, a two-dimensional culture method for cells from normal and malignant primary tissue has been described by Schlegel and colleagues. Here, we explore organoid technology to routinely establish and phenotypically annotate ‘paired organoids’ derived from adjacent tumor and healthy epithelium from CRC patients.

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Pig Taste Cell-Derived Organoids Synthesize Insulin

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