A Quantitative, Facile, and High-Throughput Image-Based Cell Migration Method Is a Robust Alternative to the Scratch Assay

被引:58
|
作者
Gough, Wendy [1 ,2 ]
Hulkower, Keren I. [4 ]
Lynch, Renee [2 ]
McGlynn, Patrick [3 ]
Uhlik, Mark [3 ]
Yan, Lei [3 ]
Lee, Jonathan A. [2 ]
机构
[1] Eli Lilly & Co, Lilly Corp Ctr, Dept Quantitat Biol, Lilly Res Labs, Indianapolis, IN 46285 USA
[2] Eli Lilly & Co, Lilly Res Labs, Dept Biol Struct, Indianapolis, IN 46285 USA
[3] Eli Lilly & Co, Lilly Res Labs, Angiogenesis Tumor Microenvironm, Indianapolis, IN 46285 USA
[4] Platypus Technol LLC, Madison, WI USA
关键词
cellular imaging; cell migration; dasatinib; high-content imaging; high-throughput screening; multiplexed assay; phenotypic assay; phenotypic mechanism of action; scratch assay;
D O I
10.1177/1087057110393340
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 T 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 T cell migration assays using an endothelial progenitor cell line and the Src kinase inhibitor dasatinib. They find that using the Acumen T Explorer laser microplate cytometer in combination with the Oris T 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. (Journal of Biomolecular Screening 2011;16:155-163)
引用
收藏
页码:155 / 163
页数:9
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