Differentiation induction of adult somatic stem cells into cardiomyocytes

成体干细胞向心肌细胞的分化诱导

• 批准号: 14370417
• 负责人: KYO Syunei
• 金额: $ 5.38万
• 依托单位: Saitama Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370417/
• 关键词: Stem_Cell; Cardiomyocyte; Regenerative Medicine; Heart Failure; Cell Transplantation

The phenomenon of regeneration is of growing interest to medical researchers. One of the object is restoration of function to a failing heart through cell transplantation, and there have been many reports seeking donor sources of stem cells, i.e. : stem cells in marrow or skeletal muscle and ES cells. In particular, reports of mesenchymal stem cell differentiation into nerve cell, myocardial cell, skeletal muscle cell, and vascular endothelial cell series have drawn attention to cell plasticity. However, it is still unclear how the stem cells can differentiate or transdifferentiate into mature cells. Moreover, we cannot maintain the stemness of the cells during cell proliferation.We examined whether gene transfer could affect both differentiation process and proliferation control. Bone marrow-derived mesenchymal stem cells, which we isolated in previous study, were transduced with Nkx2.5 and GATA4 genes that are involved in the commitment to cardiomyocytes at the initial stage. The efficacy of the in vitro differentiation to cardiomyocytes increased up to 100 fold times, compaired with non-transfectant. But, the in vivo experiment failed to demonstrate the effect of gene transfer by pathological analyses. We are thinking that the rare survival rate after cell grafting into the heart made the quantitative analyses difficult. Next, Telomerase/E6/E7/Bmi-1 were transduced into the human mesenchymal stem cells to increase the prolifelation activity. The transduced human mesenchymal stem cells proliferated over 80 population doublings without interfering with cardiomyogenic differentiation. The cells clearly exhibited differentiated cardiomyocyte phenotypes in vitro as revealed by immunocytochemistry, RT-PCR, and action potential recording. Human mesenchymal stem cells with an extended life span can be used to produce a good experimental model of cardiac cell transplantation and may serve as a highly useful cell source for cardiomyocyte transplantation.

再生现象越来越引起医学研究人员的兴趣。目的之一是通过细胞移植恢复衰竭心脏的功能，并且已经有许多报道寻求干细胞的供体来源，即：骨髓或骨骼肌中的干细胞和ES细胞。特别是，间充质干细胞分化为神经细胞、心肌细胞、骨骼肌细胞和血管内皮细胞系列的报道引起了人们对细胞可塑性的关注。然而，干细胞如何分化或转分化为成熟细胞仍不清楚。此外，我们不能保持细胞的干细胞在细胞增殖过程中，我们检查基因转移是否可以影响分化过程和增殖控制。我们在前期研究中分离的骨髓间充质干细胞中，转导了Nkx2.5和GATA 4基因，这些基因在初始阶段参与了心肌细胞的定向分化。与未转染细胞相比，转染细胞体外分化为心肌细胞的效率提高了100倍。但是，体内实验未能通过病理分析证明基因转移的效果。我们认为，细胞移植到心脏后的存活率很低，这使得定量分析变得困难。然后，将端粒酶/E6/E7/Bmi-1转导到人间充质干细胞中以增加增殖活性。转导的人骨髓间充质干细胞增殖超过80个群体，而不干扰心肌分化。通过免疫细胞化学、RT-PCR和动作电位记录显示，细胞在体外清楚地表现出分化的心肌细胞表型。具有延长寿命的人间充质干细胞可用于产生良好的心脏细胞移植实验模型，并可作为心肌细胞移植的高度有用的细胞来源。

项目成果

Yasutomi K., Y.Itokawa, H.Asada, T.Kishida, F.D.Cui, S.Ohashi, S.Gojo, Y.Ueda, T.Kubo, H.Yamagishi, J.Imanishi, T.Takeuchi, O.Mazda: "Intravascular Insulin Gene Delivery as Potential Therapeutic Intervention of the Diabetes Mellitus"Biochem Biophys. Res.

Yasutomi K.、Y.Itokawa、H.Asada、T.Kishida、F.D.Cui、S.Ohashi、S.Gojo、Y.Ueda、T.Kubo、H.Yamagishi、J.Imanishi、T.Takeuchi、O.Mazda

Yasutomi K., Y.Itokawa, H.Asada, T.Kishida, F.D.Cui, S.Ohashi, S.Gojo, Y.Ueda, T.Kubo, H.Yamagishi, J.Imanishi, T.Takeuchi, O.Mazda: "Intravascular insulin Gene Delivery as Potential Therapeutic Intervention of the Diabetes Mellitus"Biochem.Biophys.Res.Co

Yasutomi K.、Y.Itokawa、H.Asada、T.Kishida、F.D.Cui、S.Ohashi、S.Gojo、Y.Ueda、T.Kubo、H.Yamagishi、J.Imanishi、T.Takeuchi、O.Mazda

Yasutomi K., Y.Itokawa, H.Asada, T.Kishida, F.D.Cui, S.Ohashi, S.Gojo, Y.Ueda, T.Kubo, H.Yamagishi, J.Imanishi, T.Taketichi, O.Mazda: "Intravascular Insulin Gene Delivery as Potential Therapeutic Intervention of the Diabetes Mellitus"Biochem.Biophys.Res.C

Yasutomi K.、Y.Itokawa、H.Asada、T.Kishida、F.D.Cui、S.Ohashi、S.Gojo、Y.Ueda、T.Kubo、H.Yamagishi、J.Imanishi、T.Taketichi、O.Mazda

Kishida T, Asada H, Gojo S, Ohashi S, Shin-Ya M, Yasutomi K, Terauchi R, Takahashi KA, Kubo T, Imanishi J, Mazda O.: "Sequence-specific gene silencing in murine muscle induced by electroporation-mediated transfer of short interfering RNA."J.Gene Med.. 6・1

Kishida T、Asada H、Gojo S、Ohashi S、Shin-Ya M、Yasutomi K、Terauchi R、Takahashi KA、Kubo T、Imanishi J、Mazda O.：“电穿孔介导诱导的小鼠肌肉中的序列特异性基因沉默短干扰RNA的转移。“J.Gene Med..6・1

Satoshi Gojo, Akihiro Umezawa: "Plasticity of Mesenchymal Stem Cells -Regenerative Medicine for Diseased Hearts-"Human Cell. 16・1. 23-30 (2003)

Satoshi Gojo、Akihiro Umezawa：“间充质干细胞的可塑性-患病心脏的再生医学-”人类细胞16・1。