Stem Cell Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Republic of Korea.
Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology, Daejeon, Republic of Korea.
Methods Mol Biol. 2020;2126:57-71. doi: 10.1007/978-1-0716-0364-2_6.
For current and future applications of human intestinal organoids (hIOs) to various aspects of in vivo research and their potential clinical use, an efficient noninvasive system is needed to directly visualize the stage of intestinal differentiation and graft-host interactions and for further safety monitoring and efficacy. Here, we describe a detailed method for monitoring and histologically identifying implanted hIO-expressing eGFP and mCherry fluorescence under the kidney capsule of immunodeficient mice with fluorescence imaging (FLI). We then describe the orthotropic transplantation method of hIOs and methods to confirm successful engraftment in the small intestines of immunodeficient mice. These methods provide an approach for tracking the location of intestinal cells in hIOs in vivo and ex vivo using a fluorescent reporter system from the beginning of engraftment to various subsequent experiments.
为了将人类肠道类器官（hIOs）应用于体内研究的各个方面，并将其潜在的临床应用，需要一种有效的非侵入性系统来直接观察肠道分化和移植物-宿主相互作用的阶段，以及进一步的安全性监测和疗效。在这里，我们描述了一种详细的方法，用于通过荧光成像（FLI）监测和组织学识别在免疫缺陷小鼠肾包膜下表达 eGFP 和 mCherry 荧光的植入 hIO，并描述了 hIO 的异位移植方法，以及在免疫缺陷小鼠小肠中确认成功植入的方法。这些方法提供了一种使用荧光报告系统从植入开始到各种后续实验跟踪 hIO 中肠道细胞在体内和体外位置的方法。
