Interferon-responsive intestinal BEST4/CA7 cells are targets of bacterial diarrheal toxins.

BEST4/CA7 cells of the human intestine were recently identified by single-cell RNA sequencing. While their gene expression profile predicts a role in electrolyte balance, BEST4/CA7 cell function has not been explored experimentally owing to the absence of BEST4/CA7 cells in mice and the paucity of human in vitro models. Here, we establish a protocol that allows the emergence of BEST4/CA7 cells in human intestinal organoids.

Lactate controls cancer stemness and plasticity through epigenetic regulation.

Tumors arise from uncontrolled cell proliferation driven by mutations in genes that regulate stem cell renewal and differentiation. Intestinal tumors, however, retain some hierarchical organization, maintaining both cancer stem cells (CSCs) and cancer differentiated cells (CDCs). This heterogeneity, coupled with cellular plasticity enabling CDCs to revert to CSCs, contributes to therapy resistance and relapse.

Description and functional validation of human enteroendocrine cell sensors.

Enteroendocrine cells (EECs) are gut epithelial cells that respond to intestinal contents by secreting hormones, including the incretins glucagon-like peptide 1 (GLP-1) and gastric inhibitory protein (GIP), which regulate multiple physiological processes. Hormone release is controlled through metabolite-sensing proteins. Low expression, interspecies differences, and the existence of multiple EEC subtypes have posed challenges to the study of these sensors.

Intestinal organoid cocultures with microbes.

Adult-stem-cell-derived organoids model human epithelial tissues ex vivo, which enables the study of host-microbe interactions with great experimental control. This protocol comprises methods to coculture organoids with microbes, particularly focusing on human small intestinal and colon organoids exposed to individual bacterial species. Microinjection into the lumen and periphery of 3D organoids is discussed, as well as exposure of organoids to microbes in a 2D layer.

Dawn of the organoid era: 3D tissue and organ cultures revolutionize the study of development, disease, and regeneration.

Novel and updated approaches of culturing cells in 3D are rapidly advancing our understanding of development, health, and disease. As tissues have been found to behave more realistically in 3D than in 2D cultures, organoid technology in combination with recent advances in the isolation and generation of stem cells, has rapidly become a promising concept in developmental and regenerative research. The development of all kinds of tissues can now be studied "in a dish," allowing more detailed observations of stem cell maintenance, morphogens, and differentiation.