Figure 1.

Invasive breast adenocarcinoma MDA-MB-231 and MDA-MB-468 exhibited intensified focal degradation of pericellular matrix, increased invadopodia formation and poorly metastatic breast cancer MCF-7 cells were rendered invasive in presence of THP-1 macrophages. (A and B) Representative images from the in vitro matrix degradation assay. Cells (MDA-MB 231 and MDA-MB-468) were seeded on Alexa Fluor 633 labeled gelatin (Red) in absence or presence of macrophages (housed in 0.4 μm PET transwell hanging cell culture insert) and maintained for 24 h, followed by fixation, staining with Alexa fluor 488 phalloidin (Green) and mounted in aqueous media containing DAPI (Blue). Compared to mono-cultured MDA-MB-231 and MDA-MB-468 cancer cells [C], the ones that were co-cultured with macrophages [C+M] exhibited enhanced focal degradation of pericellular matrix as indicated by dark area of degraded fluorescent matrix underneath that cell. Bars represent mean invadopodia count/cell from 10 fields per experiment ±SE (*p < 0.05). (C) Compared to monocultured MCF-7 cells [C], the co-cultured MCF-7 cells (macrophages housed in 0.4 μm PET transwell hanging cell culture insert) [C+M] exhibited enhanced focal degradation (dark area of degraded fluorescent matrix underneath that cell) of pericellular matrix. Bars represent mean invadopodia count/cell (by dot count software) from 10 fields per experiment ±SE (*p < 0.05). All the experiments were done in triplicates and repeated at least thrice. Abbreviations—C: Respective cancer Cells; C+M: Respective cancer cells co-cultured with macrophages.