Gough Wendy, Hulkower Keren I, Lynch Renee, McGlynn Patrick, Uhlik Mark, Yan Lei, Lee Jonathan A
Department of Quantitative and Structural Biology, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
J Biomol Screen. 2011 Feb;16(2):155-63. doi: 10.1177/1087057110393340.
Cell migration is a key phenotype for a number of therapeutically important biological responses, including angiogenesis. A commonly used method to assess cell migration is the scratch assay, which measures the movement of cells into a wound made by physically scoring a confluent cell monolayer to create an area devoid of cells. Although this method has been adequate for qualitative characterization of migration inhibitors, it does not provide the highly reproducible results required for quantitative compound structure-activity relationship evaluation because of the inconsistent size and placement of the wound area within the microplate well. The Oris™ Cell Migration Assay presents a superior alternative to the scratch assay, permitting formation of precisely placed and homogeneously sized cell-free areas into which migration can occur without releasing factors from wounded or dead cells or damaging the underlying extracellular matrix. Herein the authors compare results from the scratch and Oris™ cell migration assays using an endothelial progenitor cell line and the Src kinase inhibitor dasatinib. They find that using the Acumen™ Explorer laser microplate cytometer in combination with the Oris™ Cell Migration Assay plate provides a robust, efficient, and cost-effective cell migration assay exhibiting excellent signal to noise, plate uniformity, and statistical validation metrics.
细胞迁移是包括血管生成在内的许多具有重要治疗意义的生物学反应的关键表型。评估细胞迁移的一种常用方法是划痕试验，该试验通过对汇合的细胞单层进行物理划痕以创建一个无细胞区域，然后测量细胞向该伤口的移动情况。尽管这种方法足以对迁移抑制剂进行定性表征，但由于微孔板孔内伤口区域的大小和位置不一致，它无法提供定量化合物构效关系评估所需的高度可重复的结果。Oris™细胞迁移试验是划痕试验的一种更好的替代方法，它允许形成精确放置且大小均匀的无细胞区域，细胞可以向其中迁移，而不会从受伤或死亡的细胞中释放因子，也不会破坏下面的细胞外基质。在本文中，作者使用内皮祖细胞系和Src激酶抑制剂达沙替尼比较了划痕试验和Oris™细胞迁移试验的结果。他们发现，将Acumen™ Explorer激光微孔板细胞仪与Oris™细胞迁移试验板结合使用，可以提供一种强大、高效且经济高效的细胞迁移试验，该试验具有出色的信噪比、板内均匀性和统计验证指标。
