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

小鼠ES细胞来源的心脏祖细胞移植对缺血性心脏的治疗效果

基本信息

批准号：
16390394
负责人：
KOMEDA Masashi
金额：
$ 8.19万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390394/
关键词：
ES cell cell transplantation old myocardial infarction myocardial regeneration Flk1 慢性心筋梗塞 FACS 急性心筋梗塞陳旧性心筋梗塞

项目摘要

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

本研究旨在探讨和建立小鼠Flk1阳性胚胎干细胞向心脏祖细胞和心肌细胞的增殖和分化过程。然后进行细胞移植，观察小鼠Flk1阳性ES细胞植入缺血心肌后的增殖和分化情况，并观察其对心功能的改善情况。研究建立小鼠Flk1阳性胚胎干细胞向心脏祖细胞和心肌细胞增殖分化的方法。1)我们建立了从小鼠Flk1阳性ES细胞单次分化和增殖300万个心肌细胞的方法。2)建立了从小鼠ES细胞中分化增殖200万个心脏祖细胞（FCV: Flk1阳性，CXCR4阳性，VE-Cadherin阴性）的方法，单次纯化纯度为80%。利用这些小鼠胚胎干细胞衍生的心脏细胞系进行细胞移植研究。在扩张型心肌病小鼠模型中，来自胚胎干细胞的心肌干细胞/祖细胞可改善心功能。2)在小鼠急性心肌梗死模型中，我们移植了来源于小鼠ES细胞的Flk1阳性心肌细胞。我们观察了供体细胞的存活，向心肌细胞的分化，抑制了梗死后左心室的扩张，增加了左心室缩短的分数。3)采用免疫缺陷小鼠建立老年心肌梗死模型。我们尝试了细胞移植治疗，但由于梗死肌壁过于薄，治疗难度太大。4)采用免疫缺陷大鼠建立老年心肌梗死模型。在心肌梗死诱导4周后，移植100万个小鼠Flk1阳性ES细胞的心脏祖细胞。首先，我们检查了移植细胞的存活和分化。细胞移植2周后处死受体大鼠，制作标本。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。我们通过超声心动图、心导管检查心功能来评价细胞移植的效果。但对改善心功能无明显作用。现在我们增加该模型的数量，并重新检查心功能的改善情况