Nanion Syncro Patch 96

描述(申请人提供)：对电可兴奋细胞的电生理学研究，如心肌细胞、骨骼肌成肌细胞、神经元、成骨细胞，对于从各种干细胞来源获得这些细胞及其在再生医学、基础生物学研究和疾病研究中的应用至关重要。例如，只有电生理测量才能确定是否已经产生了有功能的心肌细胞和神经元，因为标志物的表达是建立细胞表型所必需的，但不是充分的。这些测量需要以高速(改变镀液的短时间常数)和高通量的方式进行，以便在大的参数空间中提供关于电池属性的真实信息。这项请求是为了购买SyncroPatch 96(Nanion Technologies)，这是一种自动化的千兆密封膜片钳(APC)系统，用于研究目前可用的最高吞吐量的单个细胞的电生理特性。这一核心仪器将通过快速生成高质量的膜片钳数据，为哥伦比亚大学两个校区的七个由NIH资助的不同实验室提供先进的技术能力，以准确地表征电兴奋细胞的药理和生理特性。通过快速交换浴液(在100毫秒内)，以96孔板的形式在单个电池中快速测量全剂量响应曲线。该系统将提供哥伦比亚大学医学中心、哥伦比亚大学晨兴校区或纽约大都会地区任何其他大学目前无法提供的独特功能。如B节中对单个研究项目的详细描述，SyncroPatch 96将极大地加速我们在七个不同领域正在进行的研究：(I)高通量筛选来自携带遗传性心律失常患者的iPS-心肌细胞，(Ii)确定起搏和牵拉对来源于人iPS和胚胎干细胞(HESCs)的心肌细胞成熟的影响，(Iii)筛选治疗心力衰竭细胞内钙泄漏的新药，(Iv)调节电压依赖性钾通道；(V)确定关键的平滑肌钾通道亚单位间的结构界面，(Vi)鉴定新的遗传编码的钙通道阻滞剂，以及(Vii)通过患者来源的iPS细胞运动神经元(iPS-MN)的小分子筛选来治疗肌萎缩侧索硬化症(ALS)的新疗法。使用SyncroPatch 96收集了所有五个拟议研究领域的初步数据。该设备的所有主要用户都是经验丰富的NIH PI，在细胞响应分析方面拥有丰富的专业知识。我们希望哥伦比亚大学的调查人员和纽约市地区的合作者充分利用这一工具，促进合作。

DESCRIPTION (provided by applicant): Electrophysiological studies of electrically excitable cells, such as cardiac myocytes, skeletal myoblasts, neurons, osteoblasts, are critically important for the derivation of these cells from various stem cell sources and for their applicatio in regenerative medicine, fundamental biological research, and study of disease. For example, only electrophysiological measurements can determine if functional cardiomyocytes and neurons have been generated, as marker expression is necessary but not sufficient for the establishment of cell phenotype. These measurements need to be done at high speed (short time constants for changes of bath solutions) and in high-throughput fashion to provide realistic information about the cell properties in a large parameter space. This request is to purchase a SyncroPatch 96 (Nanion Technologies), an automated giga- seal patch clamp (APC) system for studies of electrophysiological properties of single cells with the highest throughput presently available. This core instrument would provide advanced technical capabilities to seven diverse NIH-funded laboratories across both campuses of Columbia University, by rapidly generating high quality patch clamp data for accurate pharmacological and physiological characterization of electrically excitable cells. Full dose response curves are rapidly measured in individual cells with a fast exchange of the bath solutions (within 100 ms), in a 96-well plate format. The system would provide unique capabilities not presently available either at the Columbia University Medical Center, the Columbia University Morningside Campus, or at any other University in the New York Metropolitan area. As detailed in the descriptions of the individual research projects in Section B, the SyncroPatch 96 would greatly accelerate our ongoing research in seven different areas: (i) High throughput screening of iPS-cardiomyocytes derived from patients carrying heritable arrhythmias, (ii) Determination of the impact of pacing and stretch on the maturation of cardiac myocytes derived from human iPS and embryonic stem cells (hESCs), (iii) Screening for novel drugs to treat intracellular calcium leak in heart failure, (iv) Modulation of voltage-dependent potassium channels; (v) Determination of the structural inter-subunit interface in a key smooth muscle potassium channel, (vi) Identification of new genetically encoded Ca channel blockers, and (vii) Novel therapies for amyotrophic lateral sclerosis (ALS) through small molecule screens of patient-derived iPS cell motor neurons (iPS-MNs). Preliminary data were collected using the SyncroPatch 96 in all five areas of proposed research. All Major Users of this equipment are seasoned NIH PIs with extensive expertise in analysis of cellular responses. We expect the instrument to be used at its full capacity, by investigators throughout the Columbia University and collaborators in the New York City area, promoting collaboration.