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

基本信息

  • 批准号:
    8000971
  • 负责人:
  • 金额:
    $ 7.29万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2010
  • 资助国家:
    美国
  • 起止时间:
    2010-07-01 至 2010-12-31
  • 项目状态:
    已结题

项目摘要

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):将生物相关材料高通量、平行递送和转染到细胞中仍然是一项艰巨的任务。拟议的工作通过将多个电声驱动微阵列(EAM)集成到单个 MIST(多集成样品处理)装置中来解决这个问题,该装置针对平行药物和/或基因/核酸输送进行了优化,并通过精确控制转染到多个细胞样品中生物物理作用(即同时进行的声/机械穿孔和电穿孔)。该设备格式与现有的大型细胞处理设备兼容。 EAM 是一种支持微机电系统 (MEMS) 的设备,可通过带有电穿孔电极的微观(单细胞大小)喷嘴喷射含有生物细胞的样品,从而通过机械/电穿孔组合在细胞膜上打开孔以进行吸收的纳米材料。高超声波操作频率和阵列格式能够快速处理大量细胞群(每秒高达数百万个细胞)。单个储液器系统可以容纳各种规定体积,从约 100 nL 到任意大的样本量。通过将 EAM 排列在一起,MIST 可以同时并行且均匀地处理多个不同的样品。经济的制造使得设备的流体处理组件成为一次性的,从而消除了样品交叉污染。 MIST 适合基础/应用研究以及诊断和治疗用途。 MIST 细胞处理的设计、开发和评估将建立在通过多模式台式规模原型 STEAM(通过电声驱动微阵列进行单样品处理)表征所获得的经验的基础上。对 EAM 治疗期间某些细胞类型诱导的生物效应的不断了解将有助于向多通道设备的过渡。 MIST 的开发能够同时筛选合成小干扰 RNA (siRNA) 对细胞功能的不同影响,从而有可能大大加速癌症疗法的发现。证明我们方法的可行性将为 MIST 在其他市场的商业化提供机会。 第一阶段 SBIR 的主要目标是将我们的单样本 STEAM 平台扩展到多样本 MIST 设备中,并演示多个生物分子并行输送和转染到不同的细胞样本中。为了实现这一目标,(1) 将开发针对细胞处理进行优化的 MIST 平台,(2) 将使用标准细胞评估 MIST 的活力和增殖、处理均匀性以及转染能力方面的安全处理分析、共聚焦荧光显微镜和流式细胞术。此外,MIST 治疗还将与传统的 lipofectamine 介导和电穿孔转染进行比较。 公共健康相关性:MIST(多重集成样品处理)平台的开发将满足当前对高通量、平行输送和将生物相关材料转染到细胞中的需求。此外,由于该设备格式与现有的大规模细胞处理设备(例如基于 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) 和小干扰 RNA (siRNA) 文库。使用 MIST 同时转染多个编码荧光蛋白的不同 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。

项目成果

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John Mark Meacham其他文献

John Mark Meacham的其他文献

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{{ truncateString('John Mark Meacham', 18)}}的其他基金

Acoustic platform for separation, isolation, and enrichment in biomedical research
用于生物医学研究中分离、隔离和富集的声学平台
  • 批准号:
    10445614
  • 财政年份:
    2022
  • 资助金额:
    $ 7.29万
  • 项目类别:
Acoustic platform for separation, isolation, and enrichment in biomedical research
用于生物医学研究中分离、隔离和富集的声学平台
  • 批准号:
    10681223
  • 财政年份:
    2022
  • 资助金额:
    $ 7.29万
  • 项目类别:
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
  • 财政年份:
    2010
  • 资助金额:
    $ 7.29万
  • 项目类别:
Electrosonic Ejector Microarray for Development of Cancer Therapies
用于开发癌症疗法的电声喷射器微阵列
  • 批准号:
    7611743
  • 财政年份:
    2009
  • 资助金额:
    $ 7.29万
  • 项目类别:
Electrosonic Actuation Microarray: High-Throughput Tool for Transfection of Diffi
电声驱动微阵列:用于 Diffi 转染的高通量工具
  • 批准号:
    8058167
  • 财政年份:
    2009
  • 资助金额:
    $ 7.29万
  • 项目类别:
Electrosonic Actuation Microarray: High-Throughput Tool for Transfection of Diffi
电声驱动微阵列:用于 Diffi 转染的高通量工具
  • 批准号:
    8267021
  • 财政年份:
    2009
  • 资助金额:
    $ 7.29万
  • 项目类别:

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