Bioorganic Research Institute 1 article published in JoVE Biology The 3D Culturing of Organoids from Murine Intestinal Crypts and a Single Stem Cell for Organoid Research Yuta Takase1, Kazuto Fujishima2,3, Toshio Takahashi1 1Suntory Foundation for Life Sciences, Bioorganic Research Institute, 2Institute for Integrated Cell-Material Sciences (KUIAS-iCeMS), Kyoto University, 3Faculty of Medicine, Osaka Medical and Pharmaceutical University We describe a protocol to isolate murine small intestinal crypts and culture intestinal 3D organoids from the crypts. Additionally, we describe a method to generate organoids from a single intestinal stem cell in the absence of a sub-epithelial cellular niche.