Enrichment and isolation of in vitro differentiated ES cell derived cardiomyocytes and cardiac precursor cells for transplantation into the infarcted myocardium

急性心肌梗死（AMI）导致心肌组织的不可逆损失。最近发表的数据显示人ES细胞分化为心肌细胞（CM）。基于我们自己关于使用心肌特异性MLC-2 v启动子选择来自小鼠ES细胞的CM的工作，我们现在计划通过特异性表达标记基因分离来自人ES细胞的人心室CM。工作计划旨在建立一个温和的纯化可移植CM的方案。其基于使用磁珠偶联的a-CD 4抗体的方法。在建立方案期间，嵌合CD 4-EGFP标记蛋白的细胞内EGFP部分将通过标记分子的直接可视化促进对所实现的CM纯化的评价。将对分离的细胞进行表型、电化学、电生理学和电化学表征。在使用携带实验诱导的AMI的免疫缺陷NOD/SCID小鼠的临床前研究中，将研究分离群体的功能。这包括血液动力学以及组织学和免疫组织化学分析。

Acute myocardial infarction (AMI) leads to an irreversible loss of myocardial tissue. Recently published data have shown the differentiation of human ES-cells into cardiomyocytes (CM). Based on our own work concerning the selection of CM derived from murine ES-cells using the heart-muscle specific MLC-2v promoter, we now plan to isolate human ventricular CM derived from human ES-cells via specific expression of a marker gene. The working schedule intends the establishment of a protocol for a gentle purification of transplantable CM. It is based on a method using magnetic bead coupled a-CD4-antibodies. During the establishment of the protocol the intracellular EGFP part of a chimeric CD4-EGFP marker protein will facilitate the evaluation of the achieved CM purification via direct visualisation of the marker molecule. The isolated cells will be phenotypically, immunohistochemically, electrophysiologically and pharmacologically characterised. In a preclinical study using immunodeficient NOD/SCID-mice bearing an experimentally induced AMI the isolated populations will be investigated with respect to their functionality. This comprises hemodynamic as well as histological and immunohistochemical analyses.