Isolation and Characterization of Intestinal Stem Cells

肠干细胞的分离和表征

基本信息

批准号：
7934678
负责人：
LINHENG LI
金额：
$ 35.78万
依托单位：
STOWERS INSTITUTE FOR MEDICAL RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7934678
关键词：
3-Dimensional Address Cell Proliferation Cell Separation Cell Survival Cells Colitis Controlled Environment Development Digestive System Disorders Epithelial Genes Genetic Goals Growth In Vitro Injection of therapeutic agent Intestinal Diseases Intestines Kidney Knowledge Label Life Malignant Neoplasms Membrane Membrane Proteins Methods Molecular N-Cadherin Organism Outcome Patients Plan B Play Population Preclinical Drug Evaluation Prevention Principal Investigator Proteins Public Health Regenerative Medicine Regulation Research Research Project Grants Role Signal Pathway Simulate Small Intestines Sorting - Cell Movement Stem Cell Research Stem cells Subcutaneous Tissue Surface System Therapeutic Tissues Transplantation Treatment Efficacy adult stem cell base cancer stem cell capsule cell behavior cell type cost improved in vivo insight matrigel novel public health relevance repaired self-renewal stem cell biology stem cell division tissue regeneration

项目摘要

DESCRIPTION (provided by applicant): The small intestine provides a most elegant system for the study of adult stem cells -- cells which hold great promise for regenerative medicine. Genetic studies have revealed several key signaling pathways that play a role in the regulation of intestinal stem cells (ISCs). This has provided important insight into understanding the capacity of ISCs for renewal and self-repair and differentiation into multiple intestinal cell types. However, further progress is impeded by the inability to isolate and transplant ISCs for advanced study either in vivo (within living tissue) or in vitro (within a controlled environment outside of a living organism). This research project proposes to address this barrier to ISC research, including the development of novel and reliable methods for ISC isolation, in vitro culture, and in vivo functional characterization. ISC isolation will be accomplished by using fluorescent label protein and other membrane surface proteins to sort and verify cells by genes known to be expressed in intestinal stem cells. In vitro culture will be accomplished by adding key factors that are important for supporting ISC proliferation into Matrigel or 3-dimensional culture of crypt sphere. Functional characterization will be accomplished by injecting isolated ISCs into the irradiated crypt sphere and then transplanting the chimeric sphere to an in vivo microenvironment to characterize ISC survival, self-renewal, proliferation, and lineage commitment. If this goal can be achieved, it will open avenues not only for advancing the study of ISC behavior, but also for treating intestinal disorders in which ISC-driven tissue regeneration is essential and for screening drugs to target on cancer stem cells. The ability to identify and isolate and characterize ISCs is critical for therapeutic advancements, especially tissue replacement, and for understanding cancer development, prevention and cure. PUBLIC HEALTH RELEVANCE: The cost of digestive diseases in the U.S. collectively exceeds $85 billion in direct and $23 billion in indirect expenses. Evidence indicates intestinal stem cells (ISCs) are involved in many cases -- from Crohn's to colitis to cancer. This research will increase knowledge of ISC biology, which can be applied to enhance treatment efficacies, improve patient outcomes, and ultimately, impact the public health burden of digestive diseases

描述（由申请人提供）：小肠为研究成体干细胞提供了最优雅的系统，这些细胞在再生医学方面具有广阔的前景。遗传学研究揭示了一些在肠道干细胞（ISC）调节中发挥作用的关键信号通路。这为了解 ISC 的更新、自我修复和分化为多种肠道细胞类型的能力提供了重要的见解。然而，由于无法在体内（活体组织内）或体外（在活体外部的受控环境内）分离和移植 ISC 进行高级研究，进一步的进展受到阻碍。该研究项目旨在解决 ISC 研究的这一障碍，包括开发用于 ISC 分离、体外培养和体内功能表征的新颖可靠的方法。 ISC 分离将通过使用荧光标记蛋白和其他膜表面蛋白根据已知在肠干细胞中表达的基因来分类和验证细胞来完成。体外培养将通过添加对支持 ISC 增殖重要的关键因子到基质胶或隐窝球的 3 维培养物中来完成。将分离的 ISC 注射到受辐射的隐窝球中，然后将嵌合球移植到体内微环境中，以表征 ISC 的存活、自我更新、增殖和谱系定型，从而完成功能表征。如果这一目标能够实现，它不仅将为推进 ISC 行为的研究开辟道路，而且还将为治疗肠道疾病（其中 ISC 驱动的组织再生至关重要）以及筛选针对癌症干细胞的药物开辟道路。识别、分离和表征 ISC 的能力对于治疗进展（尤其是组织替代）以及了解癌症的发展、预防和治疗至关重要。公共卫生相关性：美国消化系统疾病的直接费用总计超过 850 亿美元，间接费用超过 230 亿美元。有证据表明肠道干细胞（ISC）参与许多病例——从克罗恩病到结肠炎再到癌症。这项研究将增加对 ISC 生物学的了解，可用于提高治疗效果、改善患者的治疗效果，并最终减轻消化系统疾病的公共卫生负担