Bioengineered human colon organoids with in vivo-like complexity and function

Human intestinal organoids offer remarkable prospects for modeling intestinal homeostasis and disease. Although it is now possible to recreate many physiological features of the human gut in vitro, current approaches to generate organoids produce closed cystic structures with limited cell-type variability, hampering their relevance and applicability. Here we integrated organoid and organ-on-a-chip technology to develop a primary human stem–cell-derived organoid model, called human ‘mini-colons’ which show remarkable cell-type diversity, tissue architecture and function. Human mini-colons were maintained in culture for 14 and 21 days, and control organoids cultured in matrigel or matrigel + 20% collagen, were dissociated to single cells and analyzed by scRNAseq in three biological rep (multiplexed with hashtags) and two technical replicates (different lanes of 10XG chip).