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

摘要：患者特异性重编程体细胞（诱导多能干细胞或iPSCs）为心脏、肺和血液疾病的革命性再生治疗和药物开发应用带来了希望。例如，这些iPS细胞可以分化为心肌细胞，并且在心肌梗死后具有再生和广泛修复的能力。但分化效率低，分化的心肌细胞异质性大，主要包括成熟状态各异的心室心肌细胞。由于这些多能干细胞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不可少的。但是，建立有效的分离方法来分离分化细胞和排除导致畸胎瘤形成的细胞是将干细胞生物学的进展转化为组织替代疗法的主要挑战。传统的分离技术，如显微镜辅助人工分离是耗时的，而荧光激活细胞分选和磁激活细胞分选需要密集的劳动，外源标记或基因修饰，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了高通量电生理纯化阵列（HEPA）系统，用于根据诱导多能干细胞对电刺激的反应对其分化后代进行无标记细胞分选。第一阶段的工作

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.