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Organoids are cell-derived in vitro 3D organ models and allow the study of biological processes, such as cell behaviour, tissue repair and response to drugs or mutations, in an environment that mimics endogenous cell organisation and organ structures. Starting as a major technological breakthrough they are now firmly established as an essential tool in biological research and also have important implications for clinical use. A major advantage is that organoids can be grown from a limited supply of starting material, e.g. biopsies, and used for drug screening to develop individual therapies. They have further shown potential in the modelling of diseases, gene editing and transplantations and not least helped to answer many important biological questions. Efforts are underway to setup cryopreserved biobanks of human organoids as a resource for researchers and clinicians.
This protocol describes the generation of early-developing cardiac organoids from human pluripotent stem cells. Geometric confinement of the hiPSCs drives spatial organization of the cells from a 2D layer into 3D cardiac microchambers.
This protocol describes procedures for building the SpinΩ bioreactor for 3D tissue culture and differentiating human iPSCs into different brain region–specific organoids resembling developing human dorsal forebrain, midbrain and hypothalamus.
This protocol uses a three-layer system to organize rat primary testicular cells into organoids that can both establish and maintain germ cells in an environment containing a functional blood–testis barrier.
In this protocol, mouse or human colorectal cancer organoids are transplanted into the cecal epithelium. The transplanted organoids grow into a single primary tumor mass within the intestinal tract and tumor cells are capable of metastasis.
Colonoscopy-guided mucosal injection is used to induce site-directed tumors and to transplant tumors into the distal colons of mice. Tumors for engraftment are obtained from cancer organoids derived from mouse or human tissue and can be genetically modified before use.
This protocol describes the synthesis and application of hydrogel matrices comprising a poly(ethylene glycol) backbone, functionalized with cell adhesion cues and laminin-111. Uses include expanding stem cells and differentiating them into organoids.
This protocol describes how to recapitulate biliary development by differentiation of hPSCs into endoderm, foregut progenitor cells, hepatoblasts, cholangiocyte progenitors and mature 3D cholangiocyte-like cell organoids.
This protocol describes how to differentiate human pluripotent stem cells into nephron progenitor cells with subsequent generation of 2D and 3D kidney organoids.
This protocol describes the long-term culture of liver and pancreas 3D organoids from human and mouse, and differentiation of liver organoids in vitro and in vivo. Methodology for genetic manipulation of these self-renewing organoids is also detailed.
This protocol describes stepwise differentiation of human pluripotent stem cells into 3D kidney organoids that contain segmented nephrons connected to collecting ducts, which are surrounded by renal interstitial cells and an endothelial network.
This protocol describes a strategy for generating 3D prostate organoid cultures from healthy mouse and human prostate cells (either bulk or FAC-sorted single luminal and basal cells), metastatic prostate cancer lesions and circulating tumor cells.