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The Organoid and Cell Culture Core

类器官和细胞培养核心

基本信息

批准号：
10443137
负责人：
Hiroshi Nakagawa
金额：
$ 17.11万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-30 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10443137
关键词：
3-Dimensional Adult Bioinformatics Biological Models CRISPR screen CRISPR/Cas technology Catalogs Cell Communication Cell Culture System Cell Culture Techniques Cell Line Cells Childhood Clinical Coculture Techniques Collaborations Communities Consult Cryopreservation Data Development Digestive System Disorders Disease Disease model Doctor of Philosophy Drug Screening Educational workshop Emerging Technologies Epithelial Cells Equipment Experimental Designs Feedback Gene Expression Genes Genome Grant Growth Guide RNA Hepatobiliary Homeostasis Human Human Resources Internet Laboratories Leadership Lentivirus Vector Libraries Link Liver diseases Longevity Measures Mediating Medicine Mission Modeling Molecular Morphology Mouse Cell Line Mus Mycoplasma Mycoplasma Infections Organoids Pancreatic Diseases Patients Protocols documentation Quality Control RNA Interference Reagent Reproducibility Research Research Personnel Science Scientist Series Services Standardization Surveys System Techniques Technology Testing Tissues Training Training and Education Transgenic Mice Translational Research Translations Universities Visualization Work base bioimaging cell repository cost effective cost effective measures cost effectiveness drug testing experimental study expression vector gastrointestinal genetically modified cells high resolution imaging induced pluripotent stem cell member new technology novel operation pediatric patients pre-clinical programs repository single cell analysis tool transcriptome sequencing

项目摘要

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.

概括类器官和细胞培养核心 (OCCC) 将为哥伦比亚大学消化系的使命做出贡献和肝脏疾病研究中心（CU-DLDRC），服务会员与人类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、球体和类器官以及 3D 共培养。 OCCC 平台将使广大临床和基础 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。由博士执导。 Hiroshi Nakakawa（医学博士、博士）和 Kelley Yan（医学博士、博士）是两位长期从事研究的科学家 OCCC 将提供先进细胞培养和类器官方面的互补专业知识、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收件人。 OCCC 将提供丰富的细胞系以及人类和小鼠类器官资源库，这些资源都非常适合注释身份、传代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 研究人员从他们的人类和小鼠组织以及诱导多能干（iPS）细胞，以分析生长，通过共培养实验进行形态学、基因表达和细胞间相互作用。 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细胞和类器官系统中的干扰和诱导型逆转录病毒/慢病毒基因表达载体。这些方法通过与临床生物样本协调的多核工作流程得到进一步增强。患者和疾病特异性类器官的研究核心；具有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活动来促进 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 和国家疾病存储库特定的人类和小鼠 3D 类器官库（目标 2）；并提高新技术和新产品的技术能力通过细胞和类器官培养模型的培训建立了 CU-DLDRC 研究人员以及关于新颖发展的教育研讨会（目标 3）。 OCCC 将得到高度利用，占 CU-DLDRC 的 71% 表示使用的会员，大多数使用多种 OCCC 服务。总之，OCCC 将提供高度 CU-DLDRC 成员目前不存在的高效且具有成本效益的服务和独特模式实验室，因此构成显着的效益。通过其服务，OCCC 将促进转化 CU-DLDRC 的科学与合作，并为其促进消化疾病科学的使命做出贡献。