Novel Method for Assessing Secretory Profile of Lineage Specific Stem Cells

描述(申请人提供)：干细胞代表独特的细胞群体，具有自我更新和分化的能力。尽管“成体”干细胞可能只发育成有限数量的细胞类型，但相比之下，早期重新繁殖的干细胞有能力启动特定谱系的分化计划，并在体外形成多种细胞类型。尽管已有多项研究表明转录因子在干细胞早期分化中起重要作用，但细胞外造血细胞因子调节信号似乎更具影响力。细胞因子的产生已被广泛地应用于常规的酶联免疫吸附试验和生物测定技术，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，这使得单细胞的分泌谱的表征变得困难。利用单细胞凝胶微滴(GMD)包埋技术和流式细胞术，本SBIR旨在建立一种多参数单细胞分析方法，同时检测单个可存活的造血干/祖细胞的细胞表面标志表达和细胞因子分泌。建议的单细胞分析将提供一种方法来评估谱系特异性干细胞的分泌谱，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.