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.
