Isolation and Characterization of Intestinal Stem Cells

肠干细胞的分离和表征

• 批准号: 8332246
• 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
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8332246
• 关键词: 3-Dimensional; Address; Animal Model; Cell Proliferation; Cell Separation; Cell Survival; Cells; Colitis; Controlled Environment; Development; Digestive System Disorders; Doxycycline; Epithelial; Gene Expression Profiling; Genes; Genetic; Goals; Growth; In Vitro; Injection of therapeutic agent; Instruction; Intestinal Diseases; Intestines; Kidney; Knowledge; Label; Life; Malignant Neoplasms; Membrane; Membrane Proteins; Methods; Molecular; Organism; Outcome; Patients; Play; Preclinical Drug Evaluation; Prevention; 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; repaired; self-renewal; stem cell biology; stem cell division; tissue regeneration

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 charactize ISCs is critical for therapeutic advancements, especially tissue replacement, and for understanding cancer development, prevention and cure.

小肠为成体干细胞的研究提供了一个最优雅的系统 - 这些细胞具有很大的作用 再生医学的承诺。遗传学研究揭示了几个发挥作用的关键信号通路 肠道干细胞（ISC）的调节。这为理解 ISC 的更新、自我修复和分化为多种肠道细胞类型的能力。然而， 由于无法分离和移植 ISC 以进行体内高级研究，进一步的进展受到阻碍 （在活组织内）或体外（在活生物体外部的受控环境内）。这项研究 该项目建议解决 ISC 研究的这一障碍，包括开发新颖可靠的 ISC 分离、体外培养和体内功能表征的方法。 ISC 分离将通过使用荧光标记蛋白和其他膜表面蛋白来完成 根据已知在肠道干细胞中表达的基因对细胞进行分类和验证。体外培养将 通过将支持 ISC 增殖重要的关键因子添加到 Matrigel 或 3- 地穴球体的次元文化。功能表征将通过注入隔离的 ISC 来完成 进入受辐射的隐窝球，然后将嵌合球移植到体内微环境中 描述 ISC 生存、自我更新、增殖和谱系承诺的特征。 如果这一目标能够实现，它将不仅为推进 ISC 行为的研究开辟道路，也为 治疗肠道疾病（其中 ISC 驱动的组织再生至关重要）以及筛选靶向药物 关于癌症干细胞。识别、分离和表征 ISC 的能力对于治疗至关重要 进步，特别是组织替代，以及了解癌症的发展、预防和治疗。