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

点击翻译按钮获取中文摘要

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.