High throughput Electrophysiological Purification Array (HEPA) for cell based therapies

描述（由申请人提供）：高通量电生理净化阵列（HEPA）的细胞为基础的治疗摘要：患者特异性重编程体细胞（诱导多能干细胞或iPSC）提出了希望的革命性再生治疗心脏，肺和血液疾病和药物开发应用。例如，这些iPS细胞可以分化为心肌细胞，并具有再生能力，并在心肌梗死后进行广泛的修复。然而，分化效率低，分化的心肌细胞是异质的，主要包含具有不同成熟状态的心室心肌细胞。由于这些iPSC在体外分化成不同谱系的混合物，因此分化细胞的纯化以及它们与肿瘤形成祖细胞的分离是必需的。但是，建立有效的分离方法来分离分化的细胞并排除导致畸胎瘤形成的细胞是将干细胞生物学进展转化为组织替代疗法的主要挑战。传统的分离技术，如显微镜辅助的手动分离是耗时的，而荧光激活的细胞分选和磁激活的细胞分选需要密集的劳动、外源标记或遗传修饰，因此不容易适应临床应用。因此，为了解决这些问题，Biopico Systems与加州大学欧文分校合作开发高通量电生理纯化阵列（HEPA）系统，用于基于诱导多能干细胞及其分化后代对电刺激的反应进行无标记细胞分选。第一阶段的努力 将为获得患者特异性细胞提供基础，用于第II阶段的潜在临床应用，目标是在高活力、高通量和分选能力下获得100%特异性 用于治疗应用的多种细胞表型。为了证明所提出的平台的可行性，我们将联合收割机结合电极阵列中的基于流动的场电位感测技术与高速信号处理和高通量细胞分选，以快速检测、识别和分选数百万个特定细胞，这将允许成功地翻译细胞。 将干细胞生物学的进展应用于心脏疾病的治疗。

DESCRIPTION (provided by applicant): High throughput Electrophysiological Purification Array (HEPA) for cell based therapies Abstract: Patient-specific reprogrammed somatic cells (induced pluripotent stem cells or iPSCs) have raised hope for revolutionary regenerative treatments for heart, lung and blood diseases and drug development applications. For example, these iPS cells can be differentiated into cardiomyocytes and have the capability to regenerate and undergo extensive repair after myocardial infarction. However, the differentiation efficiency is low and differentiated cardiomyocytes are heterogeneous, containing mainly ventricular cardiomyocytes with varying maturation states. Since these iPSCs differentiate in vitro into a mixture of different lineages, purification of the differentiated cells, as well as their separatio from tumor-forming progenitors, is essential. But establishment of effective separation methods to isolate differentiated cells and exclude cells that lead to teratoma formation is the major challenge in translating advances in stem cell biology into tissue replacement therapies. Conventional separation techniques, such as microscope-assisted manual isolation is time consuming whereas fluorescence-activated cell sorting, and magnetic-activated cell sorting, require intensive labor, exogenous labeling or genetic modification and, as such, are not readily adaptable to clinical applications. To address these issues, therefore, Biopico Systems teams with the University of California, Irvine to develop High throughput Electrophysiological Purification Array (HEPA) system for label-free cell sorting of induced pluripotent stem cells and their differentiated progeny based on their response to electrical stimulation. This Phase I effort will provide the foundation for deriving patient specific cells for potential clinical use in phaseII with a goal of obtaining 100% specificity at high viability, high throughput and a capacity to sort multiple cell phenotypes for therapeutic applications. In order to demonstrate the feasibility of the proposed platform, we will combine the technologies of flow- based field potential sensing in an electrode array with high speed signal processing and high throughput cell sorting to rapidly detect, identify, and sort millions of specific cells that will allow the successful translation of advances in stem cell biology into therapies for cardiac diseases.