Multichannel Electrosonic Actuation Microarray for Cell-Based Screening

用于细胞筛选的多通道电声驱动微阵列

• 批准号: 8260892
• 负责人: John Mark Meacham
• 金额: $ 8.58万
• 依托单位: OPENCELL TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8260892
• 关键词: Multichannel; Electrosonic; Actuation; Microarray; Cell

DESCRIPTION (provided by applicant): High-throughput, parallel delivery and transfection of biologically relevant material into cells remains a difficult task. The proposed work addresses this issue through integration of multiple Electrosonic Actuation Microarrays (EAMs) into a single MIST (Multiple Integrated Sample Treatment) device that is optimized for parallel drug and/or gene/nucleic acid delivery and transfection into several cell samples via precise control of biophysical action (i.e., concurrent sono/mechanoporation and electroporation). The device format is compatible with existing large-scale cell-handling equipment. The EAM is a microelectromechanical systems (MEMS)-enabled device that ejects a sample containing biological cells through microscopic (of order size of a single cell) nozzles with incorporated electroporation electrodes, thereby opening pores in the cell membrane via combined mechano/electroporation for uptake of nanomaterials. The high ultrasonic frequency of operation and array format enable fast processing of large cell populations (up to millions of cells per second). A single reservoir system can accommodate a wide range of prescribed volumes, from ~100 nL to arbitrarily large sample sizes. By arraying EAM together, MIST enables parallel and uniform treatment of several different samples simultaneously. Economical fabrication enables fluid handling components of the device to be made disposable, which eliminates cross-sample contamination. MIST is suited to basic/applied research, as well as diagnostic and therapeutic uses. Design, development and evaluation of cell treatment by MIST will build on experience gained through characterization of a multimode bench-top-scale prototype, STEAM (Single-sample Treatment via Electrosonic Actuation Microarray). A growing understanding of the bioeffects induced in certain cell types during EAM treatment will facilitate the transition to a multichannel device. MIST development has the potential to greatly accelerate discovery of cancer therapies by enabling simultaneous screening of synthetic small interfering RNA (siRNA) for different effects on cell function. Demonstrating the feasibility of our approach will open opportunities for commercialization of MIST in other markets. The primary objective of this Phase I SBIR is to expand our single-sample STEAM platform into the multi-sample MIST device and to demonstrate parallel delivery and transfection of multiple biomolecules into different cell samples. To achieve this objective, (1) a MIST platform that is optimized for cell treatment will be developed, and (2) safe handling with regard to viability and proliferation, uniformity of treatment, and the transfection capabilities of MIST will be evaluated using standard cell assays, confocal fluorescence microscopy and flow cytometry. In addition, MIST treatment will be compared with conventional lipofectamine-mediated and electroporation-based transfection. PUBLIC HEALTH RELEVANCE: Development of a MIST (Multiple Integrated Sample Treatment) platform will address the current need for high-throughput, parallel delivery and transfection of biologically relevant material into cells. Further, because the device format is compatible with existing large-scale cell handling equipment (e.g., 384-well plate-based systems), it has the potential to accelerate discovery of potential cancer therapies through efficient screening of large complementary DNA (cDNA) and small interfering RNA (siRNA) libraries. Simultaneous transfection of multiple different DNA plasmids encoding fluorescent proteins using MIST, will demonstrate its feasibility in a wide range of drug discovery and gene therapy applications.

描述（由申请人提供）：将生物学相关材料的高通量，平行输送和转染到细胞中仍然是一项艰巨的任务。提出的工作通过将多个电音性驱动微阵列（EAM）整合到单个雾（多个集成样品处理）设备中来解决此问题，该设备可针对平行药物和/或基因/核酸递送，通过精确控制生物物理作用（即同时的Sono/机械形成和电子构造），对平行药物和/或基因/核酸的递送和转染进行了优化。设备格式与现有的大规模细胞处理设备兼容。 EAM是一种启用微机电系统（MEMS）的设备，它通过微观电动电极的微观（单个单元的大小）喷嘴弹出包含生物细胞的样品，从而通过组合的机械/电饰品在细胞膜中打开孔中的孔，以供纳米物质摄取纳米物质。高超声操作频率和阵列格式可以快速处理大型细胞种群（每秒多达数百万个单元）。单个储层系统可以容纳各种规定的体积，从〜100 nl到任意大型样本量。通过将EAM逐渐成熟，雾可以同时同时对几个不同样品进行平行和均匀处理。经济的制造使设备的流体处理组件可以使设备的易位，从而消除了跨样品污染。 薄雾适合基础/应用研究，以及诊断和治疗用途。通过薄雾的设计，开发和评估细胞处理将建立在通过表征多模式台式原型，蒸汽（通过电体驱动微阵列的单样本处理）获得的经验。对EAM治疗期间在某些细胞类型中诱导的生物效应的越来越多的理解将促进过渡到多通道设备。通过同时筛选合成小型干扰RNA（siRNA），雾气发育有可能大大加速癌症疗法的发现，从而对细胞功能产生不同的影响。证明我们方法的可行性将为其他市场的雾气商业化开放。 该阶段I SBIR的主要目的是将我们的单样本蒸汽平台扩展到多样本雾设备中，并证明将多种生物分子的平行输送和转染到不同的细胞样品中。为了实现这一目标，（1）将开发出一种针对细胞处理的雾平台，（2）将使用标准细胞测定，共焦荧光显微镜和流式细胞仪来评估有关生存能力和增殖，治疗均匀性以及雾的转染能力的安全处理。此外，将雾化处理与常规的脂肪系胺介导的和基于电穿孔的转染进行比较。 公共卫生相关性：开发雾（多重综合样品处理）平台将解决当前对生物学相关材料进行高通量，平行输送和转染细胞的需求。此外，由于设备格式与现有的大规模细胞处理设备（例如384孔板的系统）兼容，因此它有可能通过有效筛选大型互补DNA（cDNA）和小型干扰RNA（SIRNA）图书馆来加速潜在的癌症治疗方法。同时转染使用雾气编码荧光蛋白的多种不同的DNA质粒，将证明其在广泛的药物发现和基因治疗应用中的可行性。