Novel Method for Assessing Secretory Profile of Lineage Specific Stem Cells

描述（由申请人提供）：干细胞代表具有自我更新和分化能力的独特细胞群。虽然“成体”干细胞可能只发育成有限数量的细胞类型，但相反，早期再生干细胞具有启动谱系特异性分化程序并在体外形成多种细胞类型的能力。虽然一些研究已经证明，转录因子在早期干细胞分化中发挥重要作用，但细胞外造血细胞因子调节信号似乎更有影响力。已经使用常规ELISA和生物测定技术广泛研究了细胞因子的产生，然而，这些方法通常测量细胞培养上清液中的细胞因子分泌，使得难以表征单细胞的分泌谱。使用单细胞凝胶微滴（GMD）包封技术和流式细胞术，该SBIR旨在开发多参数单细胞测定法，以同时检测单个活造血干/祖细胞上的细胞表面标志物表达和细胞因子分泌。提出的单细胞测定将提供用于评估谱系特异性干细胞的分泌谱的方法，并有助于增加我们对干细胞分化的理解，用于研究和临床应用，包括细胞疗法的开发。

DESCRIPTION (provided by applicant): Stem cells represent unique cell populations that have the ability to undergo both self-renewal and differentiation. Although "adult" stem cells may develop into only a limited number of cell types, in contrast, early repopulating stem cells have the ability to initiate lineage-specific differentiation programs and to form numerous cell types in vitro. Although several studies have demonstrated that transcription factors play a significant role in early stem cell differentiation, extracellular hematopoietic cytokine regulatory signals appear to be more influential. Cytokine production has been extensively investigated using conventional ELISA and bioassay techniques, however, these methods typically measure cytokine secretion in cell culture supernatants, making characterization of secretory profiles of single cells difficult. Using single cell gel microdrop (GMD) encapsulation technology and flow cytometry, this SBIR aims to develop a multi- parameter single cell assay to detect cell surface marker expression and cytokine secretion simultaneously on single, viable hematopoietic stem/progenitor cells. The proposed single cell assay will provide a method for assessing the secretory profile of lineage specific stem cells and help to increase our understanding of stem cell differentiation for research and clinical applications including development of cellular therapies.