Endoderm Induction of Cardiac Myocytes from ES Cells

ES 细胞内胚层诱导心肌细胞

• 批准号: 7243498
• 负责人: John W Lough
• 金额: $ 32.32万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7243498
• 关键词: Address; Adult; Biochemical; Cardiac; Cardiac Myocytes; Cell Count; Cell Cycle Kinetics; Cell Death; Cell Line; Cells; Clinical; Condition; Conditioned Culture Media; Daily; Development; Disease; Effectiveness; Embryo; Embryonic Stem Cell Transplantation; Endoderm; Fibroblast Growth Factor; Fingerprint; Gene Expression; Goals; Growth Factor; Heart; Histocompatibility Testing; Human; In Situ; In Vitro; Individual; Infarction; LIF gene; Laboratories; Laboratory Finding; Membrane; Mesoderm; Methods; Modeling; Mus; Muscle Cells; Myocardial Infarction; Myocardium; Natural regeneration; Numbers; Performance; Relative (related person); Reporting; Research Personnel; Role; Serum; Survival Rate; Therapeutic; Tissues; Transplantation; Undifferentiated; Work; base; cardiogenesis; cell growth; cell type; cohort; cytokine; embryo tissue; embryonic stem cell; human embryonic stem cell; human embryonic stem cell line; improved; interest; leukemia inhibitory factor; mouse model; programs; repaired; research study; satisfaction; stem

DESCRIPTION: The potential of pluripotent embryonic stem (ES) cells to regenerate cardiac tissue has caused extraordinary interest in their therapeutic application. Clinical utilization of ES cells requires satisfaction of several issues, including how cardiac myocytes can be induced from ES cells. Toward this end, mechanisms that regulate cardiac myocyte differentiation in the embryo have provided clues. Using an approach based on earlier findings from this laboratory, we recently reported that medium conditioned by embryonic precardiac endoderm/mesoderm (E/M-cm) induces cardiogenesis in virtually all embryoid bodies (EBs) derived from mouse ES cells, and that individual EBs are >80% enriched in cardiac myocytes. The proposed work will extend these findings via performance of four Specific Aims. First, whether medium conditioned by cell-lines derived from embryonic tissues, which can be obtained in volumes sufficient for extensive biochemical and functional analysis, can mimic the cardiogenic effect of embryonic explants is being determined. It will also be determined whether medium conditioned by embryonic explants or by cell-lines derived from them can similarly induce cardiogenesis in human ES cells (line #WA01, WiCell, Madison Wl). Second, in order to identify a chemically defined cardiogenic "cocktail", medium conditioned by cardiogenic cell-lines will be characterized via a variety of cellular and biochemical methods. In the third Aim, the number and phenotypic identity of ES-derived cardiac myocytes (hereafter ES-myocytes) that are induced by conditioned medium will be precisely determined, with the objective of providing optimal cohorts for transplantation studies. Lastly, the ability of transplanted ES-myocytes to structurally and functionally repair infarcted myocardium will be evaluated in terms of differentiation state (pre-differentiated v. pluripotent), optimal donor cell numbers, proliferative potential, and survival rate. This work will help elucidate how ES cells may be utilized to repair myocardium that is damaged by disease or infarction.

描述：多能胚胎茎（ES）细胞再生心脏组织的潜力引起了对其治疗应用的非凡兴趣。 ES细胞的临床利用需要满足几个问题，包括如何从ES细胞诱导心肌细胞。为此，调节胚胎中心肌细胞分化的机制提供了线索。使用基于该实验室的早期发现的方法，我们最近报道说，由胚胎公承重性内胚层/中胚层（E/M-CM）调节的培养基在几乎所有源自小鼠ES细胞的胚胎体（EBS）中会引起心脏病，并且该单个EB在心肌肌中> 80％富集。拟议的工作将通过表现四个特定目标来扩展这些发现。首先，是否由源自胚胎组织衍生的细胞线来调节，可以在足以进行广泛的生化和功能分析的足量中获得，这是否可以模仿胚胎植体的心源性效应。还将确定是由胚胎外植体调节的培养基还是由它们得出的细胞线来诱导人类ES细胞中的心脏病发生（Line＃WA01，Wicell，Wicell，Wicell，Madison WL）。其次，为了确定一种化学定义的心源性“鸡尾酒”，将通过多种细胞和生化方法来表征以心源细胞线为条件的培养基。在第三个目标中，将精确确定由条件培养基诱导的ES衍生心肌细胞（以下称为ES-肌细胞）的数量和表型认同，目的是为移植研究提供最佳的同类群体。最后，将根据分化状态（预分化v。多能），最佳供体细胞数，增殖潜力和生存率评估，将评估受移植的ES肌细胞在结构和功能上修复梗塞心肌上的能力。这项工作将有助于阐明如何利用ES细胞来修复受疾病或梗塞破坏的心肌。