Single-Cell Stethoscopes for Functional Cardiac Cell Assessment and Sorting

 描述：诱导多能干细胞(IPSCs)的出现创造了前所未有的获得原代细胞类型的途径，如心肌细胞和神经元，这可能导致下一代药物、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不同类型的细胞混合物，需要对细胞进行分选和纯化才能进行测试或治疗。对于这些电活动的细胞，直接测试它们的电生理和功能是至关重要的 然而，目前的膜片钳测量是破坏性的，阻止了进一步的细胞使用，而且非接触式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，如作为替代的形态学或细胞表面标记。在这里，我们提出了一种新的非接触式电评估方法：“单细胞听诊器”，用于测量当动作电位激发时细胞发出的声波。这些压力波虽然很小，但可以用超灵敏的水听器进行测量。在这个项目中，我们将直接将测量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(5-10赫兹)的任何水听器更灵敏一个数量级，使测量能够向下延伸到单个细胞。我们将进一步揭示电动作电位和声学信号之间的相关性，并利用这些信号对IPSC来源的心肌细胞进行非破坏性分类。这些细胞群体将根据已知的细胞类型进行测试和基准，并对该技术的准确性进行定量评估。这项完成的技术将极大地影响IPSC来源的细胞在疾病建模、再生医学和药物发现方面的有效性和安全性。

 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.