Method for Analyzing and Isolating Stem Cells

描述（由申请人提供）：近年来，已经清楚的是，大多数细胞表面标记是由许多类型的干细胞和祖细胞共享的。因此，使用当前方法分离独特细胞亚群的能力是有限的。虽然已知干细胞分泌特定因子进入细胞外空间，但很难表征异质群体中单个活细胞的分泌活性。分泌因子与特定细胞类型的关联提供了一个额外的标记，可以用来表征细胞，并允许细胞分离，同时保持活力。结合免疫表型与分泌谱是研究和最终治疗应用的有力方法。基于分泌谱结合细胞表面抗原的表达，开发新的、可靠的方法来分析和恢复单个的、有活力的干细胞是相当有趣的。GMD干细胞分析解决了功能性、多参数、单细胞分析的需求，不损害细胞活力、生理状态和功能。第一阶段的研究优化了该检测方法，可用于几种实验性造血模型。二期研究将扩大该试验在其他模型中的应用，包括内皮祖细胞。该检测格式可广泛用于免疫学研究，包括干细胞研究和临床应用。通过提供一种可靠的方法来分析和恢复单个的、有活力的干细胞，基于分泌谱结合细胞表面抗原的表达，这种分析格式将有助于干细胞研究和临床应用。

DESCRIPTION (provided by applicant): In recent years, it has become clear that the majority of cell surface markers are shared by many types of stem and progenitor cells. Thus the ability to separate unique cell sub-populations using current methods is limited. Although it is known that stem cells secrete specific factors into the extracellular space, it is difficult to characterize secretory activity of individual viable cells within a heterogeneous population. Association of the secreted factor(s) with a specific cell type provides an additional marker that can be used to characterize the cell and permit separation of cells while maintaining viability. Combining immunophenotyping with secretory profile is powerful approach for both research and eventual therapeutic applications. Development of new, reliable methods for analyzing and recovering single, viable stem cells based on secretory profile combined with expression of cell surface antigens is of considerable interest. The GMD Stem Cell Assay addresses the need for a functional, multiparameter, single cell assay that does not compromise cell viability, physiological state, and functionality. Phase I research optimized the assay for use with several experimental hematopoietic models. Phase II research will expand use of the assay with other models, including endothelial progenitor cells. This assay format can be widely used for immunological studies, including stem cell research and clinical applications. By providing a reliable method for analyzing and recovering single, viable stem cells based on secretory profile combined with expression of cell surface antigens, this assay format will be useful for stem cell research and clinical applications.