An efficient directed cell migration assay

项目摘要 该SBIR项目的目标是设计和开发一种高效、自动化的- 亲和性定向细胞迁移(DCM)96孔法区分趋化动力学与 趋化行为。目前使用跨膜孔嵌入物来检测细胞迁移， 微流控系统和各种不同版本的划痕分析。这些方法价格昂贵，而且 提供不一致的数据。由于它在癌症和相关转移过程中的重要性， 细胞迁移被广泛研究，体外细胞迁移试验被常规用于药物。 旨在确定新的癌症疗法的发现计划。的一个重要特点 癌细胞是指它们检测化学诱导剂梯度并对其做出反应的能力。 有效筛选影响定向细胞的候选癌症治疗药物的主要障碍 迁移是指缺乏负担得起的、基于细胞的、可靠的、与生理相关的、 可重复性好，性价比高。鸭嘴兽科技有限责任公司已经开发出 将两种不同的研究细胞迁移的产品商业化。Oris“DCM”试验 在此第一阶段项目中提出的建议建立在Platypus Oris“Pro cell Migration”的成功基础上 化验产品。具体来说，Oris“DCM检测将允许测量定向细胞 对化学引诱剂作出反应的迁移，从而促进识别和发展 针对细胞对化学诱导剂的反应的先导治疗化合物。建议数 化验易于自动化，并可通过显微镜、多孔板读取器和 高内容成像器。建议的化验格式的主要其他好处包括能够 1)实时可视化细胞移动；2)执行多路传输次级屏幕，以 在同一孔内通过细胞染色阐明受试化合物的作用机制 进行原代趋化实验。后一种好处是一种非同寻常的优势， 通过节约试剂增加研究经费的知识生成能力 包括化合物、细胞等，以及以其他方式重复消耗的资源 竞争格式要求的测试。

Project Summary The goal of this SBIR project is to design and develop an efficient, automation- compatible Directed Cell Migration (DCM) 96-well Assay that distinguishes chemokinetic from chemotactic behavior. Cell migration is currently assayed using trans-membrane well inserts, microfluidic systems and various versions of scratch assays. These methods are expensive and provide inconsistent data. 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. An important characteristic of cancer cells is their ability to detect chemoattractant gradients and migrate in response to them. A major barrier to efficient screening of candidate cancer therapeutics affecting directed cell migration is the lack of affordable cell based assays that are robust, physiologically relevant, reproducible and cost-effective to perform. Platypus Technologies LLC has developed and commercialized two distinct products for the study of cell migration. The Oris" DCM assay proposed in this Phase I project builds on the success of the Platypus Oris" Pro cell migration assay product. Specifically, the Oris" DCM assay will allow measurement of directed cell migration in response to chemoattractants, thereby facilitating identification and development of lead therapeutic compounds that target cellular responses to chemoattractants. The proposed assay is amenable to automation and to readout by microscopy, multi-well plate readers, and high content imagers. Major additional benefits of the proposed assay format include ability to 1) visualize cell movement in real-time and 2) perform multiplexed secondary screens to elucidate mechanism of action of the test compound via cytostaining within the same well that the primary chemotaxis assay was run. This latter benefit is an extraordinary advantage that increases the knowledge-generating power of the research dollar by conserving reagents including compounds, cells, etc and resources that would otherwise be consumed in repetitive testing required by competing formats.