In-vitro generation of cardiomyocytes from MAPC

MAPC 体外生成心肌细胞

• 批准号: 6833335
• 负责人: WOUTER J VAN'T HOF
• 金额: $ 10万
• 依托单位: ATHERSYS, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6833335
• 关键词: bioassay; cardiac myocytes; cardiogenesis; cell differentiation; cell growth regulation; cell population study; cell proliferation; cell transplantation; drug discovery /isolation; flow cytometry; gene expression; genetically modified animals; green fluorescent proteins; growth factor; high throughput technology; laboratory mouse; mature animal; method development; pluripotent stem cells

DESCRIPTION (provided by applicant): The development and institution of stem cell and tissue engineering methodologies hold the potential to target many of the most significant cardiovascular diseases and have a dramatic impact on the treatment of one of the leading causes of death in the United States healthcare system. The current proposal is a collaborative effort between Athersys, Inc. and the University of Minnesota Stem Cell Institute within their cell therapy program. This project has a main focus on exploiting the Multipotent Adult Progenitor Cell (MAPC) technology as a novel platform for the in-vitro generation of cardiomyocytes for potential application in allogeneic cell therapy in heart disease. The overall objective of this proposal is to synergize the small molecule compound high-throughput screening (HTS) capability at Athersys with the MAPC platform. The working hypothesis is that this provides a unique opportunity to identify factors and pathways that can initiate specific and controlled differentiation of MAPCs into cardiomyocytes or progenitors. The Phase I effort will focus on the use of MAPCs acquired from transgenic mouse models in which the green fluorescent protein (GFP) reporter is specifically expressed in cardiac muscle through the activity of cardiac muscle specific promoter sequences. MAPCs will be isolated from these transgenic mice and expanded for use in HTS assays that measure the induction of GFP fluorescence as a quantitative read-out for differentiation of MAPCs into the cardiomyocyte lineage. The two specific aims in Phase I are; 1) to isolate and expand MAPC from transgenic mice with cardiac muscle specific expression of GFP, and; 2) to establish a sensitive and specific assay for quantification of MAPC differentiation into the cardiomyocyte lineage based on GFP fluorescence. The ultimate ambition of this proposal is to establish the conditions for ex vivo production of cardiomyocyte progenitors from mouse as well as human MAPCs by the end of Phase II. This will provide clinically and commercially relevant new sources for cardiac muscle cells, which are in great demand for in vitro toxicology studies as a near term product, or for the further development of allogeneic cellular therapies for heart disease.

描述（由申请人提供）：干细胞和组织工程方法的开发和机构具有靶向许多最重要的心血管疾病的潜力，并对美国医疗保健系统中主要死亡原因之一的治疗产生巨大影响。目前的提案是Athersys，Inc.和明尼苏达大学干细胞研究所在他们的细胞治疗计划。该项目的主要重点是利用多能成体祖细胞（MAPC）技术作为体外产生心肌细胞的新平台，用于心脏病的同种异体细胞治疗。本提案的总体目标是使Athersys的小分子化合物高通量筛选（HTS）能力与MAPC平台协同。工作假设是，这提供了一个独特的机会，以确定因素和途径，可以启动特定的和控制分化的MAPCs的心肌细胞或祖细胞。第一阶段的努力将集中在使用从转基因小鼠模型中获得的MAPC，其中绿色荧光蛋白（GFP）报告基因通过心肌特异性启动子序列的活性在心肌中特异性表达。将从这些转基因小鼠中分离MAPC并扩增以用于HTS测定，HTS测定测量GFP荧光的诱导作为MAPC分化成心肌细胞谱系的定量读数。I期的两个具体目标是：1）从具有GFP的心肌特异性表达的转基因小鼠中分离和扩增MAPC，以及2）建立用于基于GFP荧光定量MAPC分化成心肌细胞谱系的灵敏且特异的测定。该提议的最终目标是在II期结束时建立从小鼠以及人MAPC离体产生心肌细胞祖细胞的条件。这将为心肌细胞提供临床和商业上相关的新来源，这些心肌细胞作为近期产品在体外毒理学研究中或进一步开发心脏病的同种异体细胞疗法中有很大的需求。