Stem Cell/Organoid and Genome Editing Core

项目摘要 消化健康中心（DHC）多能干细胞和类器官核心的总体目标是 为DHC研究人员提供全面的综合服务，以促进对消化机制的研究， 通过使用天然和基因组编辑的干细胞和类器官的创新和可靠的模型系统来治疗疾病。 核心通过四个相辅相成的目标来实现这一目标。在第一个目标中，“为DHC调查人员提供 人类iPSC和iPSC衍生的类器官和培训，“核心向用户提供质量测试， 人多能干细胞（PSC）和PSC衍生的食管、胃底、胃肠道和胃肠道的多谱系细胞类器官 以及胃、肠、结肠和肝的胃窦区域。核心还促进了组织的产生- 衍生的类肠和肝和胆类器官。这些是新颖的3D微型器官， 与人类相关的生理学和病理学。核心领导和经验丰富的员工持有 通过正式的分层课程和实践培训、监督和支持进行技术转让。在 第二个目标是“为更便宜，可扩展，高质量的类器官提供下一代类器官技术”。 核心为研究人员提供了从健康或患病受试者中产生类器官的机会 使用新的、可靠的方案来更有效和可扩展地将人PSC定向分化为 类器官在第三个目标“为CRISPR-Cas9介导的人类PSC基因组编辑提供服务”中， 核心在技术和服务范围上不断发展，为DHC研究人员提供最先进的基因组 使用CRISPR/Cas9系统进行编辑。成功的项目使用敲入和敲除编辑来研究 疾病的机制。为了能够在实验测定中跟踪和可视化这些细胞，Core还 提供标记的PSC编辑和筛选服务，以建立基于 调查人员的需要。第四个目标是“储存并提供质量受控的iPSC生产线， 用于消化研究的类器官，”核心库良好的表型天然和编辑的hPSC系，以及 疾病特异性iPSCs，以催化基础研究成果转化为新的诊断和 治疗工具该服务符合高质量的指标，并符合法规要求， 身份识别和人类受试者保护。总的来说，这些新技术使DHC调查人员能够 使用多细胞实验系统研究疾病的机制，这些系统与正常细胞有直接的“谱系”， 和患病的人体组织（包括在不同的成熟阶段）。服务的提供是 精简和集中，并定位调查人员探索新的合作项目，以促进 这些发现将对消化系统疾病研究产生积极影响。

PROJECT SUMMARY The overall goal of the Pluripotent Stem Cell and Organoid Core of the Digestive Health Center (DHC) is to provide DHC investigators with fully integrated services to catalyze research on mechanisms of digestive diseases via innovative and reliable model systems using native and genome edited stem cells and organoids. The Core pursues this goal with four complementary aims. In the First Aim “to provide DHC investigators with human iPSCs and iPSC-derived organoids and training,” the Core makes available to users quality-tested human pluripotent stem cells (PSCs) and PSC-derived multilineage cellular organoids of the esophagus, fundic and antral regions of the stomach, intestine, colon, and liver. The Core also facilitates the production of tissue- derived enteroids and liver and biliary organoids. These are novel 3D-miniature organs that enable studies related to physiology and pathobiology relevant to humans. Core leaders and experienced staff hold technology transfer through formal tiered courses and hands-on training, supervision, and support. In the Second Aim “to provide next generation organoid technologies for cheaper, scalable, high-quality organoids,” the Core provides investigators with the opportunity to generate organoids from healthy or diseased subjects using novel, reliable protocols for more efficient and scalable directed differentiation of human PSCs into organoids. In the Third Aim “to provide services for CRISPR-Cas9 mediated genome editing of human PSCs,” the Core evolved in its scope of technology and service to offer DHC Investigators state-of-the-art genome editing using CRISPR/Cas9 system. Successful projects have used knockin and knockout editing to study mechanisms of disease. To be able to track and visualize these cells in experimental assays, the Core also provide reporter-labeled PSC-editing and screening services to establish novel cell lines based on investigators’ needs. And in the Fourth Aim “to bank and make available quality-controlled iPSC lines and organoids for digestive research,” the Core banks well phenotyped natural and edited hPSC lines as well as disease-specific iPSCs to catalyze the translation of basic research findings into novel diagnostic and therapeutic tools. The service is aligned with high quality metrics and meets regulatory requirements for de- identification and human subject protection. Collectively, these novel technologies empower DHC investigators to study mechanisms of disease using multi-cellular experimental systems that have direct “lineage” to normal and diseased human tissues (including at different stages of maturation). The delivery of services is streamlined and centralized, and positions investigators to explore new collaborative projects to catalyze discoveries that will positively impact digestive disease research.