The Organoid and Cell Culture Core

摘要 器官和细胞培养核心(OCCC)将为哥伦比亚大学消化中心的使命做出贡献 和肝病研究中心(CU-DLDRC)，服务于成员与人和小鼠细胞相关的需求 用于胃肠道、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、球体和有机体以及3D联合培养。OCCC 平台将使大多数临床和基础CU-DLDRC研究人员受益，为研究提供强大的工具 上皮细胞及其在消化稳态和疾病中的相互作用，这是CU-DLDRC的中心主题。 由中川浩博士和严凯利博士执导，这两位科学家长期从事 作为先进细胞培养和有机化合物方面的补充专业知识，OCCC将提供服务、技术、 CU-DLDRC调查员的质量控制和成本效益，包括试点和可行性补助金 收件人。OCCC将提供丰富的细胞系和人类和小鼠器官类物质 对身份、通道编号和支原体无感染状态进行注释，从而提供质量控制， 严密性和可重复性。OCCC还将协助CU-DLDRC调查人员从 他们的人类和小鼠组织以及可诱导的多能干细胞(IPS)，以分析生长情况， 通过共培养实验的形态、基因表达和细胞与细胞的相互作用。OCCC将整合 通过CRISPR/Cas9、RNA调控基因组和基因表达的先进细胞培养技术 干扰和诱导逆转录病毒/慢病毒基因在细胞和有机物系统中的表达载体。 这些方法通过与临床生物医学和生物医学中心协调的多核心工作流程得到进一步增强 针对患者和疾病特定有机物的研究核心；具有生物信息学和单细胞分析核心 用于基于平板RNAseq的CRISPR和药物筛选；以及用于功能可视化的生物成像核心 细胞和细胞器。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和国家疾病储存库-- 特定的人和鼠3D有机类库(目标2)；并加强新的和 通过细胞和有机物培养模型的培训建立了CU-DLDRC调查人员，以及 关于新发展的教育研讨会(目标3)。OCCC将得到高度利用，CU-DLDRC的利用率将达到71% 会员表示使用，其中大部分使用多项OCCC服务。总之，OCCC将提供高度的 高效、经济实惠的服务和独特的模式，这是CU-DLDRC成员目前不存在的 实验室，因此构成了重大的好处。通过其服务，OCCC将促进翻译 该中心致力于促进该中心的科学和合作，并为其促进消化疾病科学的使命作出贡献。

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.