MDA-MB-231 cell morphology influences chemotactic sensing of CXCL12 gradients in type 1 bovine collagen matrix

Chemotaxis plays a critical role in the metastatic progression of breast cancer. The chemokine CXCL12 is well recognized as an essential component of chemotactic migration in triple-negative breast cancer (TNBC) cells in vivo . The purpose of this study is to determine how the highly metastatic TNBC cell line, MDA-MB-231, migrates in response to well-defined CXCL12 gradients in vitro . Traditional 2D transwell migration assays were optimized to gauge the MDA-MB-231 cells’ responsiveness to various CXCL12 concentrations. The optimum chemoattractant concentrations were applied to a commercially available 3D chemotaxis assay as stable linearly diffused gradients. Cells were embedded in type 1 bovine collagen at two different collagen concentrations, and individual unlabeled cells were monitored for 24 hours using brightfield microscopy. Time-lapse videos were used to track cell movement and shape. Quantitative data analysis was performed using an automated tracking software to measure chemotactic parameters based on cell morphology. MDA-MB-231 cells were responsive to CXCL12 concentrations greater than 200 ng/mL in 2D and 3D systems. In 3D systems, significant directed migration was observed in denser collagen matrices. It was observed that in 3D matrices a range of cell morphologies was present. Therefore, chemotaxis was evaluated as a function of cell shape revealing some differences between sub cellular populations. Our findings show the cell’s shape influences the chemotactic sensing towards CXCL12 gradients.