An Efficient Fully 3-Dimensional Cell Migration Assay

高效的全 3 维细胞迁移测定

• 批准号: 8713184
• 负责人: Timothy George Burland
• 金额: $ 14.74万
• 依托单位: PLATYPUS TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2015-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8713184
• 关键词: 3-Dimensional; Affect; Award; Behavior; Biochemical; Biocompatible; Biological Assay; Biological Sciences; Biomimetics; Cell Culture Techniques; Cell Line; Cell physiology; Cells; Characteristics; Chemicals; Collagen; Collagen Type I; Cues; D Cells; Data; Deposition; Detection; Development; Dimensions; Ensure; Environment; Exhibits; Extracellular Matrix; Figs - dietary; Gel; Goals; Health; Image; In Vitro; Knowledge; Laboratories; Lead; Malignant Neoplasms; Measurement; Membrane; Methods; Microfluidics; Microscope; Microscopy; Migration Assay; Monitor; Ornithorhynchus anatinus; Phase; Phenotype; Physiological; Predictive Value; Procedures; Proteins; Qualifying; Reader; Reagent; Research; Research Personnel; Resources; Robotics; Small Business Innovation Research Grant; Specificity; Staging; Staining method; Stains; Surface; System; Technology; Testing; Therapeutic; Time; Toxicology; Water; base; cell motility; cell type; cellular targeting; cost effective; design; drug discovery; flexibility; improved; in vivo; inhibitor/antagonist; innovation; instrumentation; metastatic process; migration; novel therapeutics; polymerization; programs; promoter; public health relevance; response; screening; success; surface coating; technological innovation; two-dimensional

DESCRIPTION (provided by applicant): The goal of this SBIR project is to design and develop an efficient, physiologically relevant and cost-effective 96-well 3-D Cell Migration Assay that is compatible with mid-high throughput/high content assay platforms. Due to its importance in cancer and associated metastatic processes, cell migration is intensively studied and in vitro cell migration assays are routinely used in drug discovery programs aimed at identifying new cancer therapeutics. Cell migration studies of cells grown in 3-D biomimetic matrices are currently performed primarily using trans-membrane well inserts and microfluidic systems. These methods are expensive, laborious and provide inconsistent data. Moreover, these assays compromise on an important characteristic of the in vivo environment: the unique spatial arrangement of cells that facilitates their ability to exhibit normal physiological phenotypes and functions. A major barrier to efficient screening of candidate cancer therapeutics affecting cell migration is the lack of affordable cell based assays that are robust, physiologically relevant, reproducible and cost-effective to perform. The Oris" 3-D migration assay proposed in this Phase I project builds on the successful Oris" Pro cell migration assay product that Platypus developed and commercialized with the support of a prior SBIR award. The Oris" 3-D assay will allow monitoring of cell migration in a more closely in vivo-like microenvironment, thereby facilitating identification and development of lead therapeutic compounds that target cellular responses. The proposed assay is fully automatable and is compatible with readout by microscopy, multi-well plate readers, and high content imagers. Major additional benefits of the proposed assay include ability to i) provide cells with a fully 3-D microenvironment in the entire duration of the migration assay; (ii) use a universal system for both adherent and non-adherent cells; iii) visualize cell movement in real-time and iv) perform multiplexed secondary screens to elucidate mechanism of action of the test compound via cytostaining within the same well. This latter benefit is an extraordinary advantage that increases the knowledge-generating power of the research dollar by conserving reagents, cells, and other resources that would otherwise be consumed in the repetitive testing required by competing formats.

描述（由申请人提供）：该SBIR项目的目标是设计和开发一种有效，生理相关且具有成本效益的96孔3-D细胞迁移测定法，该测定与中高吞吐量/高含量分析平台兼容。由于其在癌症和相关转移过程中的重要性，细胞迁移经过深入研究和体外细胞 迁移测定通常用于旨在识别新癌症治疗剂的药物发现计划中。目前，主要使用跨膜井插入物和微流体系统进行的3-D仿生基质中生长的细胞的细胞迁移研究。这些方法昂贵，费力，并提供不一致的数据。此外，这些测定在体内环境的重要特征上妥协：细胞的独特空间排列促进其表现出正常的生理表型和功能的能力。有效筛查影响细胞迁移的候选癌症治疗剂的主要障碍是缺乏可负担的基于可负担的细胞测定，这些测定能够稳健，生理上相关，可重现和成本效益。在此阶段I项目中提出的ORIS“ 3-D迁移测定法基于成功的Oris” Pro细胞迁移分析产品，Platypus在先前的SBIR奖励的支持下开发和商业化了。 ORIS“ 3-D分析将允许在更紧密的体内微环境中监测细胞迁移，从而促进靶向细胞反应的铅治疗化合物的识别和开发。所提出的测定完全可自动化。与Microscopy一起提供了与Mirti Moscopy的读数相兼容，并提供了三台读取者的读数。在整个迁移分析的持续时间（ii）中的微环境； iii均使用通用系统来实时可视化细胞的细胞运动，并阐明多重细胞的多种筛选机制，从以及其他资源，这些资源将在竞争格式所需的重复测试中消耗。