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.

项目摘要-核心B 离体3-D“微型肠”培养系统，称为肠样，由人类外科手术产生。 已经发现，在组织标本或内窥镜活组织检查中， 包括正常肠上皮的隐窝-绒毛轴。发展这些 肠杆菌提供了一个令人兴奋的机会，以促进我们的理解，如何致病 以及有益的微生物，其中许多可以防止或减轻严重性 病原体引起的肠道疾病，与肠道相互作用并诱导肠道反应， 上皮因此，人类肠样核心的长期目标是（1） 表征和比较从小肠的三个区域产生的人类肠 （十二指肠，空肠，回肠）和结肠，从不同的病人建立其作为模型 人类肠道疾病，（2）测试新的方法，以改善肠道作为一个相关的 正常肠上皮的模型，和（3）提供这些肠样组织和特化生长 用于本申请中拟定的其他项目（核心C，项目1、2和3）的试剂。 人类肠样核心的目标是提供一个集中的设施，以满足肠样 参与NAMSED提案的所有研究人员的需求。的具体职能 核心将有两个功能：一个服务组件，将（1）生产肠样培养物 从我们现有的所有项目库中提取（2）从选定的新患者样本中建立肠样蛋白 (3)维持所有肠状组织的冷冻库存（4）根据请求通过并区分肠状组织 (5)维持WNT、头蛋白和r-spondin产生细胞系，并进行标准化条件培养。 用于肠样细胞培养的培养基（6）对肠样细胞系的分化状态进行细胞培养试验（7） 通过培训小组成员进行技术转让;开发部分，将（1） 制定和标准化生理指标作为肠样反应的指标（2）修改 （3）共培养，以增强或消除特定细胞类型或基因的存在 间充质细胞来测试生长因子的替代，并开发新的结构 组件（例如，肠胚倒置，使顶面朝外）（4） 整合免疫细胞以提高肠模型的相关性。芯会 响应个别项目的需求，这可能会随着研究项目的变化而变化。 随着整个油田的发展和项目3中新平台的设计，新 将开展活动，以满足我们和其他NASMED项目调查人员的需求。 我们的目标是互补和合作，在这些令人兴奋的努力，发展肠 作为研究肠道疾病的模型。