Authors

A Ward¹; J Trigg¹; K Barnes¹; G Lovell¹; T Dale¹; 
¹ Sartorius, UK

Discussion

Cell migration is a multistep process that enables cell movement in response to an environmental stimulus, this contributes to physiological and pathological processes, including wound healing and tumor metastasis. The rapid rearrangement of actin filaments leads to a cycle of leading edge protrusion and lagging edge retraction, summating in the cellular morphological and kinetic changes for migration. Here we exemplify a robust assay for the visual assessment of cell migration and its modulation.

Incucyte® Live-Cell Analysis enables automated image-based measurements of cell migration in vitro via label-free or dual fluorescence readouts. Cells were seeded, incubated until confluent, wounded and treated with compounds. To simultaneously monitor cell cycle dynamics, cells expressing the Incucyte® Cell Cycle Lentivirus reagent were used. Images were acquired using the Incucyte® Live-Cell Analysis System and analyzed using integrated software. In HT-1080 fibrosarcoma cells and MDA-MB-231 breast cancer cells, the pharmacological profiles of PP242 (mTOR inhibitor) and cytochalasin D (actin polymerization inhibitor) were determined. Cytochalasin D caused complete inhibition of migration in both cell lines, whereas PP242 exerted a greater antimigratory effect on MDA-MB-231 than HT-1080 cells (57% vs 98% Relative Wound Density, respectively at 1.25 µM after 24h). For HT-1080 cells treated with 0.31 µM cytochalasin D, we found inhibition of migration and cell cycle where a high percentage of cells in the wound are in G1 phase (red) compared to vehicle (86% vs 45% Red population, respectively at 16h). Data is consistent with research as cytochalasin D arrests the cell cycle at the G1-S transition via activating p53-dependent pathways.

This data exemplifies that live-cell analysis, utilizing reproducible methods and intuitive analysis software, is a high throughput approach to quantify cell migration, which is amenable to a range of cell types and pharmacological agents