Human Enteroid Core

人肠类核心

• 批准号: 8855933
• 负责人: NOAH Freeman SHROYER
• 金额: $ 15.06万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-15 至 2020-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8855933
• 关键词: 3-Dimensional; Animal Model; Apical; Biological Models; Categories; Cause of Death; Cell Line; Cells; Cessation of life; Child; Coculture Techniques; Colon; Conditioned Culture Media; Development; Disease; Duodenum; Endoscopic Biopsy; Engineering; Ensure; Enteral; Face; Freezing; Functional disorder; Genes; Goals; Growth; Growth Factor; Human; Immune; Immune system; Individual; Intestines; Measures; Mesenchymal; Mesenchyme; Modeling; Modification; Operative Surgical Procedures; Patients; Physiological; Physiology; Population; Pre-Clinical Model; Reagent; Reproducibility; Research Personnel; Research Project Grants; Sampling; Services; Severities; Small Intestines; Specimen; Surface; System; Technology Transfer; Testing; Tissues; Training; Validation; Villus; aged; cell type; cost effective; gastrointestinal epithelium; human disease; ileum; improved; intestinal epithelium; jejunum; meetings; member; microorganism; novel; novel strategies; pathogen; prevent; response; three-dimensional modeling

Project Summary – Core B Ex-vivo 3-D “mini-intestine” culture systems, termed enteroids, generated from human surgical specimens or endoscopic biopsies, have been found to recapitulate the multiple cell types that comprise the crypt-villus axis of the normal intestinal epithelium. The development of these enteroids has provided an exciting opportunity to advance our understanding of how pathogenic as well as beneficial microorganisms, many of which can prevent or lessen the severity of pathogen-induced diarrheal illness, interact with and induce responses from the intestinal epithelium. Therefore, the long range objectives of the Human Enteroid Core are to (1) characterize and compare human enteroids generated from three regions of the small intestine (duodenum, jejunum, ileum) and colon, from different patients to establish their use as models of human diarrheal diseases, (2) test new approaches to improve the enteroids as a relevant model of normal intestinal epithelium, and (3) to provide these enteroids and specialized growth reagents for use in the other projects (Core C, Projects 1, 2, and 3) proposed in this application. The goal of the Human Enteroid Core is to provide a centralized facility to meet the enteroid needs of all of the investigators involved in the NAMSED proposal. The specific functions of the Core will have two functions: a Service component, which will (1) Produce enteroid cultures from our existing bank for all projects (2) Establish enteroids from select new patient samples (3) Maintain frozen stocks of all enteroids (4) Passage and differentiate enteroids per requests (5) Maintain WNT, noggin, and r-spondin producing cells lines and standardized conditioned media for enteroid culture (6) Routinely test the enteroid lines for differentiation status (7) Technology transfer through training group members; a Development component, which will (1) Develop and standardize physiological measures as indicators of enteroid responses (2) Modify enteroids to enhance or deplete the presence of specific cell types or genes (3) Co-culture with mesenchymal cells to test for substitution of growth factors and develop novel structural components (e.g., inversion of the enteroids such that the apical surface faces outward) (4) Incorporate immune cells to improve the relevance of the enteroid model. The Core will be responsive to needs of the individual Projects, which may change as the research projects proceed, as the overall field evolves and as new platforms are engineered in Project 3. New activities will be developed to meet the needs of our and other NASMED project investigators. Our goal is complementary and collaborative in these exciting efforts to develop the enteroids as models to study enteric disease.

项目总结 – 核心 B 离体 3-D“迷你肠”培养系统，称为肠类，由人体手术产生 标本或内窥镜活检，已被发现概括了多种细胞类型 包括正常肠上皮的隐窝绒毛轴。这些的发展 Enteroids 提供了一个令人兴奋的机会来加深我们对致病性的理解 以及有益微生物，其中许多可以预防或减轻严重程度 病原体引起的腹泻疾病，与肠道相互作用并诱导反应 上皮。因此，人类肠类核心的长期目标是（1） 表征并比较小肠三个区域产生的人类肠类物质 来自不同患者的（十二指肠、空肠、回肠）和结肠，以建立它们作为模型的用途 人类腹泻病的研究，(2) 测试改善肠类相关疾病的新方法 正常肠上皮模型，以及（3）提供这些肠样和专门的生长 用于本申请中提出的其他项目（核心 C、项目 1、2 和 3）的试剂。 人类肠类核心的目标是提供一个集中设施来满足肠类 参与 NAMSED 提案的所有研究人员的需求。具体职能 核心将具有两个功能：一个服务组件，它将 (1) 产生肠菌培养物 从我们现有的银行中用于所有项目 (2) 从选择的新患者样本中建立肠类 (3) 维持所有 Enteroid 的冷冻库存 (4) 根据请求传代和区分 Enteroid (5) 维持WNT、noggin和r-spondin产生细胞系并标准化条件化 用于肠样培养的培养基 (6) 定期测试肠样细胞系的分化状态 (7) 通过培训小组成员进行技术转让；开发组件，它将 (1) 开发和标准化生理测量作为肠样反应的指标 (2) 修改 肠类增强或消除特定细胞类型或基因的存在 (3) 与 间充质细胞测试生长因子的替代并开发新的结构 组件（例如，肠体反转，使顶端表面朝外）(4) 合并免疫细胞以提高肠样模型的相关性。核心将是 响应各个项目的需求，这些需求可能会随着研究项目的变化而变化 随着整个领域的发展以及项目 3 中新平台的设计，我们将继续前进。 新 将开展活动以满足我们和其他 NASMED 项目研究人员的需求。 我们的目标是在这些令人兴奋的开发肠类药物的努力中进行互补和协作 作为研究肠道疾病的模型。