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
- 来源:
- 关键词:AddressBiocompatible MaterialsBiologicalBiological AssayBiological SciencesBiologyCell LineCell ProliferationCell SizeCell membraneCell physiologyCellsCellular biologyCharacteristicsChemicalsClinicalCollaborationsColorComplementary DNADNADNA deliveryDU145DataDevelopmentDevice or Instrument DevelopmentDevicesDiagnosticElectrodesElectroporationElementsEmbryoEquipmentEvaluationExhibitsFlow CytometryFluorescence MicroscopyFoundationsFrequenciesFutureGenesGreen Fluorescent ProteinsHumanIndividualInvestigationLaboratoriesLifeLipofectamineLiquid substanceManufacturer NameMarketingMechanicsMediatingMethodsMicroscopicNucleic AcidsPerformancePharmaceutical PreparationsPhasePlasmidsPolymersPopulationPositioning AttributePreparationPreventionProcessProteinsPumpRNA libraryReagentResearchResearch InstituteRobotRoboticsSample SizeSamplingScreening procedureSmall Business Innovation Research GrantSmall Interfering RNASystemTechniquesTechnologyTherapeutic UsesTimeTransducersTransfectionUltrasonicsUniversitiesVariantWorkbasecancer therapycell typeclinical applicationcommercializationconventional therapydesigndrug discoveryexperiencefluorexongene therapyhigh throughput screeninghuman diseaseimprovedkidney cellnanomaterialsnoveloperationplasmid DNAprototypepublic health relevancesample collectionsealsmall moleculesonoporationtherapy developmenttooluptake
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):将生物学相关材料高通量、平行递送和转染到细胞中仍然是一项艰巨的任务。所提出的工作通过将多个电声致动微阵列(EAM)集成到单个MIST(多个集成样品处理)装置中来解决这个问题,该MIST装置被优化用于通过精确控制生物物理作用(即,同时声/机械穿孔和电穿孔)。该设备格式与现有的大规模细胞处理设备兼容。 EAM是微机电系统(MEMS)使能的装置,其通过具有并入的电穿孔电极的微观(单个细胞的量级大小)喷嘴来激发含有生物细胞的样品,从而经由组合的机械/电穿孔来打开细胞膜中的孔以摄取纳米材料。高超声操作频率和阵列格式能够快速处理大细胞群(每秒高达数百万个细胞)。单个储液器系统可容纳范围广泛的规定体积,从约100 nL到任意大的样品量。通过将EAM排列在一起,MIST可以同时并行和统一处理几个不同的样品。经济的制造使得装置的流体处理部件能够被制成一次性的,这消除了交叉样品污染。 MIST适用于基础/应用研究以及诊断和治疗用途。MIST的细胞处理的设计、开发和评估将建立在通过表征多模式台式规模原型STEAM(通过电声致动微阵列进行单样本处理)获得的经验基础上。对EAM治疗期间某些细胞类型中诱导的生物效应的不断了解将促进向多通道设备的过渡。MIST的开发有可能通过同时筛选合成的小干扰RNA(siRNA)对细胞功能的不同影响来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品STEAM平台扩展到多样品MIST设备,并展示多个生物分子平行递送和转染到不同的细胞样品中。为了实现这一目标,(1)将开发一种针对细胞处理进行优化的MIST平台,以及(2)将使用标准细胞测定、共聚焦荧光显微镜和流式细胞术评价MIST在活力和增殖、处理均匀性和转染能力方面的安全处理。此外,将MIST治疗与常规脂质体介导的和基于电穿孔的转染进行比较。
公共卫生相关性:MIST(多重集成样品处理)平台的开发将满足当前对高通量、平行递送和转染生物相关材料进入细胞的需求。此外,由于装置格式与现有的大规模细胞处理设备(例如,384-基于孔板的系统),其具有通过有效筛选大互补DNA(cDNA)和小干扰RNA(siRNA)文库来加速发现潜在癌症疗法的潜力。使用MIST同时转染编码荧光蛋白的多个不同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
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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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
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