Evaluation system of cardiac cells and tissues with combined micro-electrode array based electrical mapping and optical mapping system

基于组合微电极阵列的电测绘和光学测绘系统的心脏细胞和组织评估系统

• 批准号: 16200033
• 负责人: SAKUMA Ichiro
• 金额: $ 31.62万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 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/en/grant/KAKENHI-PROJECT-16200033/
• 关键词: micro-electrode array; optical mapping; regenerative medicine; arrhythmia; 細胞培養; 光学測定; 電極アレイ; 電気刺激; 再生医療

For progress of regenerative medicine in cardiology, such as bio-pacemaker based on cultured cardiac tissue for arrhythmia therapy, we need a system to evaluate function of cardiac cell, cardiac tissue, and a whole heart where the regenerated tissue is implanted. In this study, combined optical and electrical measurement system of cultured cardiac tissue and whole heart preparation has been developed.1) We have developed a cardiac electric stimulation system with real time feedback of action potential propagation dynamics. In this system, cardiac electrical propagation is visualized by optical mapping method with high spatial and temporal resolution. Propagated excitation wave front is determined in real time, and the location and timing of electrical stimulation is controlled depending on the propagation pattern. It can initiate Make/Break excitation by controlling the timing and location considering the phase (depolarized, repolarized, and resting phase) of the cardiac tissue. The function of the system was confirmed by experiments with Langendorff perfused rabbit heart preparations.2) We have also developed measurement system for cultured cardiac cells and stem cells where temporal and spatial electrical activation patterns are measured by an integrated micro-electrode arrays fabricated with photo lithography (64 channels, 30 μm X 30 μm, electrode spacing: 130 μm), and a fluorescence image of intracellular Ca is measured by a cooled CCD camera. Cardiac cells were separated from neonatal rats and cultured. mechanical contraction was observed after three days in vitro. It grows to contraction of extended areas of cultured cells. Synchronization of the electrical activities and mechanical contraction were also identified. Intracellular calcium concentration was measured by fluorescent dye (fluo-4) simultaneously. Synchronization of calcium dynamics and electrical activities were identified.

为了心脏病学再生医学的进展，例如基于培养心脏组织的生物起搏器用于心律失常治疗，我们需要一个系统来评估心脏细胞、心脏组织和植入再生组织的整个心脏的功能。本研究开发了体外培养心脏组织和全心制备的光电联合测量系统。1)研制了具有动作电位传播动态实时反馈的心脏电刺激系统。该系统采用高时空分辨率的光学成像方法，实现了心电传播的可视化。实时确定传播激励波前，并根据传播方向控制电刺激的位置和时间。它可以根据心脏组织的相（去极化、再极化和静息相），通过控制时间和位置来启动Make/Break激励。用兰根多夫灌注兔心脏制剂验证了该系统的功能。2)我们还开发了用于培养心脏细胞和干细胞的测量系统，其中通过光刻技术制造的集成微电极阵列（64通道，30 μm X 30 μm，电极间距：130 μm）测量时间和空间电激活模式，并通过冷却CCD相机测量细胞内Ca的荧光图像。将新生大鼠心肌细胞分离培养。体外3天后观察机械收缩。它生长为培养细胞的扩展区域的收缩。电活动和机械收缩的同步也被确定。同时用荧光染料（fluo-4）测定细胞内钙浓度。确定了钙动力学和电活动的同步性。

项目成果

Why Do Cardiac Arrhythmias Occur?--Experimental, In-Silico and Clinical Approach--

为什么会发生心律失常？--实验、计算机模拟和临床方法--

• 发表时间: 2006
• 期刊: CORONA PUBLITHING CO.,LTD.
• 作者: Hiroshi Inada;Itsuo Kodama;Ichiro Sakuma;Kazuo Nakazawa
• 通讯作者: Kazuo Nakazawa

心筋細胞活動電位シュミレーションを用いたQT延長メカニズムの検討

使用心肌细胞动作电位模拟检查 QT 延长的机制

• 发表时间: 2005
• 期刊: 生体医工学 43(in press)
• 作者: 山口;荒船;児玉;佐久間他
• 通讯作者: 佐久間他

光学マッピングにおける心臓興奮伝播の動態解析

使用光学映射对心脏兴奋传播进行动态分析

• 发表时间: 2005
• 期刊: 生体医工学 43
• 作者: ヘルランバンニコラス;高本眞一;土肥健純;他;Hongen Liao;Yutaka Tanizawa;Hongen Liao;廖 洪恩;ヘルランバン ニコラス;Yuhei Takata;Hongen Liao;Keri Kim;Keri Kim;Hongen Liao;D.Kim;A.Gooya;H.Yamashita;Akihiko Yagi;Nicholas Herlambang;J.Hong;Hongen Liao;徳国淳一;松居和広;ヘルランバン ニコラス;浦野克紀;八木昭彦;山下紘正;山下紘正;森川茂廣;小原 太;徳田淳一;ヘルランバン ニコラス;野口雅史;Masafumi Noguchi;Masafumi Noguchi;野口雅史;吉田大樹;高田祐平
• 通讯作者: 高田祐平

心筋細胞活動電位数値計算モデルを用いた動物種によるIKrブロッカーに対する感受性の違いに関する検証

使用心肌细胞动作电位数值计算模型验证不同动物种类对 IKr 阻滞剂的敏感性差异

• 发表时间: 2006
• 期刊: 生体医工学 44(in press)
• 作者: 山口;荒船;佐久間;寺澤;柴田;本荘;児玉;神谷
• 通讯作者: 神谷

The state of the art of nanobioscience in Japan

日本纳米生物科学的最新水平

• 发表时间: 2006
• 期刊: IEEE Transactions on Nanobioscience 5・1
• 作者: Eguchi Y;et al.;Sachiko Yamaguchi et al.;Takako Saotome et al.;Shoogo Ueno et al.
• 通讯作者: Shoogo Ueno et al.