Fig. 7
Three-dimensional collagen invasion assays and quantification of cell phenotypes of invasive inflammatory ductal carcinoma (IFDUC)1 mammary epithelial cells (MEC) and mesenchymal cells (MES) incubated with conditioned medium (CM) from different adipose-derived stem cell (ADSC)-differentiated adipocyte stages. IFDUC1-MEC and IFDUC1-MES cells were incubated with CM from ADSC-IFDUC1 and ADSC-normal mammary cells (NORMA)4 during adipocyte differentiation at the time points 7, 14 and 21 days to test the influence on invasion for 72 h. Y-axis represents total number of invaded IFDUC1-MEC cells (invaded cells/cm2); X-axis represents ADSC-CM for control (−− = no CM), and CM at 7, 14 and 21 days; gray bar represents CM from ADSC-IFDUC1 and black bar from ADSC-NORMA4. Directly below three CM dilutions (1:10, 1:20 and 1:30) were tested and correspond with specific days (a). Examples of IFDUC1-MEC cells on the collagen surface (top) and invaded mesenchymal or amoeboid phenotypes below the collagen surface (middle and bottom), supporting epithelial to mesenchymal transition (EMT) or epithelial to amoeboid transition; bar = 50 μm (b). Y-axis represents total number of invaded IFDUC1-MES cells (invaded cells/cm2); X-axis represents ADSC-CM for control (−− = no CM), and CM at 7, 14 and 21 days; gray bar represents CM from ADSC-IFDUC1 and black bar from ADSC-NORMA4. Directly below three CM dilutions (1:10, 1:20 and 1:30) were tested and correspond with specific days (c, d). IFDUC1-MES cells on the collagen surface (left) and from the same field invaded cells below the collagen surface; invaded cells are out of focus (left panels) but become clear when focusing below the surface (right panels). Bar = 50 μm (e)