An Efficient Fully 3-Dimensional Cell Migration Assay

描述(申请人提供)：该SBIR项目的目标是设计和开发一种与中高通量/高含量分析平台兼容的高效、生理相关和成本效益高的96孔3-D细胞迁移检测方法。由于它在癌症及其相关转移过程中的重要性，细胞迁移受到了广泛的研究，体外细胞 迁移分析通常用于旨在识别新的癌症疗法的药物发现计划中。细胞在三维仿生基质中生长的细胞迁移研究目前主要使用跨膜孔板和微流控系统进行。这些方法昂贵、费力，而且提供的数据不一致。此外，这些分析还损害了体内环境的一个重要特征：细胞独特的空间排列，有助于它们表现出正常的生理表型和功能。有效筛选影响细胞迁移的候选癌症治疗方法的一个主要障碍是缺乏负担得起的、基于细胞的、可靠的、生理上相关的、可重复性的和成本效益高的检测方法。在此第一阶段项目中提出的Oris“3-D迁移分析建立在成功的Oris”Pro细胞迁移分析产品的基础上，该产品由Platypus开发并在之前的SBIR奖的支持下商业化。Oris“3-D测试将允许在更接近体内的微环境中监控细胞迁移，从而促进针对细胞反应的先导治疗化合物的识别和开发。建议的分析是完全自动化的，并与显微镜、多孔板阅读器和高含量成像仪的读数兼容。建议的测试的主要其他好处包括：i)在整个迁移测试过程中为细胞提供完全三维的微环境；(Ii)对贴壁和非贴壁细胞使用通用系统；iii)实时显示细胞运动；以及iv)执行多路二次筛选，通过同一孔内的细胞染色来阐明测试化合物的作用机制。后一种好处是一个非同寻常的优势，它通过节省试剂、细胞和其他资源来增加研究资金的知识产生能力，否则这些资源将在竞争格式所需的重复测试中消耗。

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.