Single Cell Migration Assay Using Human Breast Cancer MDA-MB-231 Cell Line

Bio Protoc. 2020 Apr 20;10(8):e3586. doi: 10.21769/bioprotoc.3586.

Abstract

Cell migration is a fundamental cellular process that plays a crucial role in many physioglogical and pathological processes such as wound healing or cancer metastasis. Many assays have been developed to examine cell migration, such as the wound healing or scratch assay, Boyden Chamber or transwell assay, and the method we will describe here, single cell migration assay. In this assay, cells are plated sparsely on a collagen coated plate and live cell imaging is performed over a period of 2 h at 1 frame per minute. After imaging is completed, cells are tracked manually using ImageJ by tracking movement of the centroid of the cell. These data points are then exported and overall distance travelled from frame to frame is determined and divided by total time imaged to determine speed of the cell. This method provides a quick way to examine effect of cellular manipulation on cell migration before proceeding to perform more complex assays.

Keywords: 2D migration; Cell tracking; Live cell imaging; Random migration; Single cell migration.