Single-Cell Stethoscopes for Functional Cardiac Cell Assessment and Sorting

用于功能性心肌细胞评估和分选的单细胞听诊器

• 批准号: 9327802
• 负责人: NICHOLAS A MELOSH
• 金额: $ 23.7万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-15 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9327802
• 关键词: Ablation; Acoustics; Actinin; Action Potentials; Alpha Cell; Architecture; Behavior; Benchmarking; Cardiac; Cardiac Myocytes; Cardiology; Cell Culture Techniques; Cell Separation; Cell physiology; Cell surface; Cells; Collection; Devices; Disease model; Effectiveness; Electrophysiology (science); Fire - disasters; Frequencies; Head; Heart Diseases; Individual; Ion Channel; Lasers; Lead; Light; Measurement; Measures; Membrane; Mental Depression; Methods; Morphology; Nanotechnology; Neurons; Pharmaceutical Preparations; Phenotype; Population; Process; Proxy; Regenerative Medicine; Research; Resolution; Safety; Scanning; Scientist; Signal Transduction; Sorting - Cell Movement; Stethoscopes; Techniques; Technology; Testing; Therapeutic; Time; Troponin T; Tympanic membrane; base; blebbistatin; cancer therapy; cell type; drug discovery; experimental study; human embryonic stem cell; induced pluripotent stem cell; next generation; novel; patch clamp; pressure; prevent; programs; public health relevance; screening

 DESCRIPTION: The advent of induced pluripotent stems cells (iPSCs) has created unprecedented access to primary cell types such as cardiomyocytes and neurons, which may lead to the next-generation of drugs, regenerative medicine, and cancer treatments. Unfortunately, the iPS transformation and differentiation process produces a heterogeneous mixture of cell types, required cell sorting and purification for testing or therapy. For these electrically active cells it is critical to directly test their electrophysiological and functional behavior, yet current patch-clamping measurements are destructive, preventing further cell use, and non-contact methods do not have sufficient resolution to discriminate between different phenotypes. The field is thus limited to non-functional analysis techniques such as morphology or cell-surface markers as proxies. Here we propose a new method for non-contact electrical assessment: "Single Cell Stethoscopes" for measuring the acoustic waves given off by the cell when an action potential fires. While small, these pressure waves are measureable with ultra-sensitive hydrophones. Under this program, we will directly correlate the measured acoustic signals to patch clamp electrophysiology, and demonstrate the ability to identify individual cardiomyocyte cell types. These hydrophones will be an order of magnitude more sensitive than any existing at the relevant frequency ranges for cardiomyocytes (5-10 Hz), enabling measurements down to individual cells. We will further reveal the correlation between the electronic action potential and acoustic signals, and use these to non-destructively categorize and iPSC derived cardiomyocytes. These cell populations will be tested and benchmarked against known cell types, and the accuracy of the technique quantitatively assessed. This completed technology will dramatically impact the effectiveness and safety of iPSC-derived cells for disease modeling, regenerative medicine, and drug discovery.

 描述：诱导多能茎细胞（IPSC）的进步已经创造了对主要细胞类型的前所未有的访问，例如心肌细胞和神经元，这可能导致药物的下一代，再生医学和癌症治疗。不幸的是，IPS的转化和分化过程产生了细胞类型的异质混合物，所需的细胞分选和纯化进行测试或治疗。对于这些电活动电池，直接测试其电生理和功能至关重要 行为，但是当前的斑块夹测量是破坏性的，防止了进一步的细胞使用，非接触方法没有足够的分辨率来区分不同的表型。因此，该领域仅限于非功能分析技术，例如形态学或细胞表面标记作为代理。在这里，我们提出了一种用于非连接电评估的新方法：当动作电势发射时，用于测量细胞发出的声波的“单细胞听诊器”。虽然很小，但这些压力波是用超敏感的氢气测量的。在此程序下，我们将直接将测得的声学信号与贴片夹电物质生理学相关联，并证明鉴定单个心肌细胞类型的能力。这些言语将比在相关频率范围（5-10 Hz）的相关频率范围内的任何数量级更敏感，从而使测量值降低到单个细胞。我们将进一步揭示电子动作电位和声学信号之间的相关性，并将其用于非破坏性分类和IPSC衍生的心肌细胞。这些细胞群体将针对已知的细胞类型进行测试和基准测试，并定量评估该技术的准确性。这项完成的技术将极大地影响IPSC衍生细胞在疾病建模，再生医学和药物发现的有效性和安全性。