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高效、生理相关且具有成本效益的96孔3-D细胞迁移试验，该试验与中高通量/高含量试验平台兼容。由于其在癌症和相关转移过程中的重要性，细胞迁移被深入研究，并且在体外细胞迁移中， 迁移分析通常用于旨在鉴定新的癌症治疗剂的药物发现计划中。细胞迁移的细胞生长在3-D仿生矩阵的研究，目前主要使用跨膜孔插入和微流体系统。这些方法都是昂贵的，费力的，并提供不一致的数据。此外，这些测定在体内环境的一个重要特征上妥协：细胞的独特空间排列，这有助于它们表现出正常生理表型和功能的能力。影响细胞迁移的候选癌症治疗剂的有效筛选的主要障碍是缺乏可负担得起的基于细胞的测定，所述测定是稳健的、生理学相关的、可再现的和成本有效的。Oris第一阶段计划中的3D细胞迁移分析仪是在Oris Pro细胞迁移分析仪的基础上开发的，Oris Pro细胞迁移分析仪是Platypus在SBIR奖项的支持下开发并商业化的。Oris的3D分析将允许在更接近体内的微环境中监测细胞迁移，从而促进识别和开发针对细胞反应的领先治疗化合物。所提出的测定是完全自动化的，并且与通过显微镜、多孔板读取器和高含量成像器的读出兼容。所提出的测定法的主要额外益处包括以下能力：i）在迁移测定法的整个持续时间内为细胞提供完全3-D微环境;（ii）使用用于粘附和非粘附细胞的通用系统; iii）实时可视化细胞移动和iv）进行多重二次筛选以通过同一孔内的细胞染色阐明测试化合物的作用机制。后一个好处是一个非凡的优势，它通过节省试剂、细胞和其他资源来增加研究资金的知识生成能力，否则这些资源将被消耗在竞争形式所需的重复测试中。