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

用于基于细胞的高通量电生理净化阵列 (HEPA)

• 批准号: 8787704
• 负责人: John Collins
• 金额: $ 24.73万
• 依托单位: BIOPICO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-21 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8787704
• 关键词: Address; Adverse effects; Algorithms; Amplifiers; Asses; Biological Assay; California; Cardiac Myocytes; Cell Separation; Cell Therapy; Cells; Cellular Morphology; Clinical; Clinical Laboratory Techniques; Computer software; Databases; Derivation procedure; Detection; Development; Differentiation Antigens; Disease; Electric Stimulation; Electrodes; Feasibility Studies; Fluorescence-Activated Cell Sorting; Fluorescent Dyes; Foundations; Gene Expression; Genetic; Geometry; Glass; Goals; Health; Heart Diseases; Height; Hematological Disease; Human; In Vitro; Individual; Label; Laboratories; Lead; Lung diseases; Magnetism; Manuals; Measurement; Mechanics; Methods; Metric; Microfluidics; Microscope; Modification; Morphologic artifacts; Myocardial Infarction; Natural regeneration; Noise; Patients; Performance; Phase; Phenotype; Protocols documentation; Pump; Regenerative Medicine; Replacement Therapy; Response to stimulus physiology; Risk; Sampling; Signal Transduction; Somatic Cell; Sorting - Cell Movement; Specificity; Speed; Stem cells; Sterility; Stimulus; Syringes; System; Techniques; Technology; Teratoma; Therapeutic; Time; Tissues; Transgenes; Translating; Translations; Undifferentiated; Universities; Validation; Ventricular; Width; abstracting; base; clinical application; design; drug development; electric impedance; epigenetic marker; fluorexon; human disease; induced pluripotent stem cell; novel; pluripotency; prevent; progenitor; prototype; regenerative; remyelination; repaired; research study; response; sensor; signal processing; simulation; stem cell biology; stem cell differentiation; tool; tumor; verification and validation

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.

描述（由申请人提供）：用于细胞治疗的高通量电生理纯化阵列 (HEPA) 摘要：患者特异性重编程体细胞（诱导多能干细胞或 iPSC）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，并且具有再生能力并在心肌梗塞后进行广泛的修复。然而，分化效率低且分化的心肌细胞异质，主要包含成熟状态不同的心室心肌细胞。由于这些 iPSC 在体外分化成不同谱系的混合物，因此纯化分化细胞以及将其与肿瘤形成祖细胞分离至关重要。但建立有效的分离方法来分离分化细胞并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，例如显微镜辅助的手动分离非常耗时，而荧光激活细胞分选和磁激活细胞分选则需要大量劳动力、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以用于 II 期潜在临床应用奠定基础，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，我们将把电极阵列中基于流的场电位传感技术与高速信号处理和高通量细胞分选相结合，以快速检测、识别和分选数百万个特定细胞，从而成功翻译 干细胞生物学在心脏病治疗中的进展。