The Organoid and Cell Culture Core

总结 类器官和细胞培养核心（OCCC）将有助于哥伦比亚大学消化系统的使命 和肝脏疾病研究中心（CU-DLDRC）通过服务成员的需求与人类和小鼠细胞 用于胃肠、肝胆和胰腺疾病建模的培养系统。这些系统包括 原代培养、基因工程细胞系、球状体和类器官以及3D共培养。OCCC 平台将使大多数临床和基础CU-DLDRC研究者受益，为研究提供强大的工具 上皮细胞及其在消化系统内稳态和疾病中的相互作用，CU-DLDRC的中心主题。 由Hiroshi Nakagawa博士，医学博士，博士和Kelley Yan博士，医学博士，博士，两位科学家长期以来 OCCC将在先进细胞培养和类器官方面提供互补的专业知识，提供服务、技术， CU-DLDRC研究人员的质量控制和成本效益，包括试点和可行性补助金 受惠人士OCCC将提供丰富的细胞系和人类和小鼠类器官库， 注释鉴别、传代次数和无支原体感染状态，从而提供质量控制， 严谨性和可重复性。OCCC还将协助CU-DLDRC研究人员从 它们的人类和小鼠组织以及诱导性多能干细胞（iPS），以分析生长， 形态学、基因表达和细胞-细胞相互作用。OCCC将整合 通过CRISPR/Cas9、RNA操纵基因组和基因表达的先进细胞培养技术 干扰，以及细胞和类器官系统中的诱导型逆转录病毒/慢病毒基因表达载体。 这些方法通过与临床生物样本和 针对患者和疾病特异性类器官的研究核心;生物信息学和单细胞分析核心 用于基于板RNAseq的CRISPR和药物筛选;以及用于功能可视化的生物成像核心 细胞和类器官。OCCC将通过以下相互关联的方式促进CU-DLDRC的使命 具体目标：为CU-DLDRC成员及其实验室提供细胞系、先进的细胞培养 平台和技术（目标1）;建立和扩大独特的CU-DLDRC和国家疾病资料库， 特异性人类和小鼠3D类器官文库（Aim 2）;以及增强新的和 通过对细胞和类器官培养模型的培训，建立了CU-DLDRC研究人员， 关于新发展的教育研讨会（目标3）。OCCC将得到高度利用，CU-DLDRC将达到71% 会员表示使用，他们中的大多数人使用多种OCCC服务。总而言之，OCCC将提供高度的 高效和具有成本效益的服务以及CU-DLDRC成员目前不存在的独特模式 实验室，因此具有很大的优势。通过其服务，OCCC将促进翻译 在CU-DLDRC的科学和合作，并有助于其使命，以促进消化系统疾病的科学。

SUMMARY The Organoid and Cell Culture Core (OCCC) will contribute to the mission of the Columbia University Digestive and Liver Disease Research Center (CU-DLDRC) by serving members’ needs related to human and murine cell culture systems for modeling of gastrointestinal, hepatobiliary and pancreatic diseases. These systems include primary culture, genetically-engineered cell lines, spheroids and organoids as well as 3D co-culture. OCCC platforms will benefit the majority of clinical and basic CU-DLDRC investigators, providing powerful tools to study epithelial cells and their interactions in digestive homeostasis and disease, the central theme of the CU-DLDRC. Directed by Drs. Hiroshi Nakagawa, MD, PhD and Kelley Yan, MD, PhD, two scientists with long-standing complementary expertise in advanced cell culture and organoids, the OCCC will provide services, technologies, quality control and cost-effectiveness to the CU-DLDRC investigators, including the Pilot and Feasibility grant recipients. The OCCC will provide a rich repository of cell lines and human and mouse organoids that are well annotated for identity, passage number, and Mycoplasma infection-free status, thereby providing quality control, rigor and reproducibility. The OCCC will also assist CU-DLDRC investigators to generate 3D organoids from their human and mouse tissues as well as from inducible pluripotent stem (iPS) cells, to analyze growth, morphology, gene expression, and cell-cell interactions via co-culture experiments. The OCCC will integrate advanced cell culture technologies for manipulating genome and gene expression via CRISPR/Cas9, RNA interference, and inducible retroviral/lentiviral gene expression vectors in cell and organoid-based systems. These approaches are further enhanced by coordinated multi-core workflows with the Clinical Biospecimen and Research Core for patient- and disease-specific organoids; with the Bioinformatics and Single Cell Analysis Core for the plate-RNAseq-based CRISPR and drug screens; and with the Bioimaging Core for functional visualization of cells and organoids. The OCCC will promote the mission of the CU-DLDRC through the following interrelated Specific Aims: To provide CU-DLDRC members and their laboratories with cell lines, advanced cell culture platforms and technologies (Aim 1); to build and expand unique CU-DLDRC and national repositories of disease- specific human and mouse 3D organoid libraries (Aim 2); and to enhance the technical capabilities of new and established CU-DLDRC investigators through training on cell and organoid culture models and as well as educational seminars on novel developments (Aim 3). The OCCC will be highly utilized with 71% of CU-DLDRC members indicating use, most of them use of multiple OCCC services. In summary, the OCCC will provide highly efficient and cost-effective services and unique models that do not currently exist in CU-DLDRC members’ laboratory and therefore constitute significant benefits. Through its services, OCCC will stimulate translational science and collaborations in the CU-DLDRC and contribute to its mission to promote digestive disease science.