The therapeutic efficacy of mouse ES cell derived cardiac progenitor cell transplantation in the ischemic heart

本研究旨在探索并建立Flk 1阳性小鼠ES细胞向心肌祖细胞和心肌细胞分化的方法。然后，本研究通过细胞移植的方法，观察小鼠Flk 1阳性ES细胞移植到缺血心肌后的增殖分化情况，并观察其对心功能的改善情况，探讨并建立小鼠Flk 1阳性ES细胞向心肌祖细胞和心肌细胞增殖分化的方法。建立了从Flk 1阳性的ES细胞一次分化增殖300万心肌细胞的方法（FCV：Flk 1阳性、CXCR 4阳性、VE-Cadherin阴性），纯度达80%以上，进行了小鼠ES细胞来源的心肌细胞系的细胞移植研究：1）Flk 1阳性、CXCR 4阳性、VE-Cadherin阴性; 关于我们 胚胎干细胞来源的心脏干/祖细胞改善扩张型心肌病小鼠模型的心功能。2）在小鼠急性心肌梗死模型中，移植来源于胚胎干细胞的Flk 1阳性心肌细胞。结果：1.在此模型中，供体细胞存活率高，分化为心肌细胞，心肌梗死后左室扩张明显减少，左室短轴缩短率增加。我们尝试了细胞移植治疗，但由于梗死肌壁过薄，治疗难度较大。4）采用免疫缺陷大鼠建立陈旧性心肌梗死模型。诱导心肌梗死4周后，移植100万个来自小鼠Flk 1阳性ES细胞的心脏祖细胞，首先检测移植细胞的存活和分化情况，细胞移植2周后，处死受体大鼠并制成标本。我们通过GFP染色和向心肌细胞的分化证实了大量移植细胞的存活，通过检查心脏功能如超声心动图和心导管检查来评估细胞移植的效果。但对心功能的改善作用不明显，现增加该模型的数量，复查心功能改善情况。

This study was to investigate and establish the procedure to proliferate and differentiate the mouse Flk1 positive ES cells to cardiac progenitor cells and cardiomyocytes. Then, cell transplantation was done to investigate the proliferation and differentiation of mouse Flk1 positive ES cells after implanted into ischemic myocardium and to check the improvement of cardiac function.To investigate and establish the procedure to proliferate and differentiate the mouse Flk1 positive ES cells to cardiac progenitor cells and cardiomyocytes.1) We established the procedure to differentiate and proliferate 3million cardiomyocytes from mouse Flk1 positive ES cells at single approach.2) We established the procedure to differentiate and proliferate the 2million cardiac progenitor cells (FCV : Flk1 positive, CXCR4 positive, VE-Cadherin negative) from mouse ES cells, with the purity of 80% at single approach.Cell transplantation study using these mouse ES cell derived cardiac cell line.1) Flk1 positi … More ve cardiac stem/progenitor cells derived from embryonic stem cells improve cardiac function in a dilated cardiomyopathy mouse model.2) In the mouse acute myocardial infarction model, we transplanted Flk1 positive cardiomyocytes which derived from mouse ES cells. We identified the donor cells survivality, differentiation into cardiomyocytes, restrain the left ventricle dilatation after infarction, and increase the fractional shortening in this model.3) We performed the old myocardial infarction model using immunodeficiency mouse. We tried the cell transplantation therapy, but it was too difficult to do the therapy, due to the excessive thinness of the infarcted muscle wall.4) We performed the old myocardial infarction model using immunodeficiency rat. 4 weeks after the induction of myocardial infarction, we transplanted 1million cardiac progenitor cells from mouse Flk1 positive ES cell.First, we examined the transplanted cells survival and differentiation.2 weeks after cell transplantation, recipient rats were sacrificed and specimens were made. We confirmed a large mass of transplanted cells survivajity by GFP staining and the differentiation to cardiomyocytes.We evaluated the effect of cell transplantation by checking the cardiac function such as echocardiography and cardiac catheterization. But, there was no apparent effect of improvement of cardiac function.Now we increasing the number of this model, and rechecking the improvement of cardiac function Less