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：诱导多能干细胞 (iPSC) 的出现为心肌细胞和神经元等原代细胞类型提供了前所未有的途径，这可能会导致下一代药物、再生医学和癌症治疗。分化过程产生细胞类型的异质混合物，需要细胞分选和纯化以进行测试或治疗。对于这些电活性细胞，直接测试其电生理和功能至关重要。 但目前的膜片钳测量具有破坏性，阻碍了细胞的进一步使用，并且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。在这里，我们提出了一种非接触式电评估的新方法：“单细胞听诊器”，用于测量动作电位激发时细胞发出的声波，这些压力波虽然很小，但可以用超灵敏的方法测量。根据该计划，我们将直接将测量到的声学信号与膜片钳电生理学相关联，并展示识别单个心肌细胞类型的能力，这些水听器将比现有的心肌细胞相关频率范围敏感一个数量级。 （5-10 Hz），能够测量到单个细胞，我们将揭示电子动作电位和声信号之间的相关性，并使用它们对 iPSC 衍生的心肌细胞进行非破坏性分类。该技术以已知细胞类型为基准，并进行了定量评估，其准确性将极大地影响 iPSC 衍生细胞在疾病建模、再生医学和药物发现方面的有效性和安全性。