Fig. 3

Myoepithelial cells (MEPs) reduce invasive phenotype of ductal carcinoma in situ (DCIS) structures formed in mammary architecture and microenvironment engineering (MAME) cultures. a MCF10.DCIS-lenti-RFP (DCIS) were seeded in MAME cultures alone or with N1ME cells (MEPs) and imaged live at day 16. Left columns are tiled images of 16 contiguous differential interference contrast (DIC) fields. Scale bar = 100 μm. Middle columns are magnified DIC images of the boxed areas in the left columms and illustrate invasive outgrowths from DCIS structures in the absence of MEPs (outlined area and arrow in top row, middle column). Scale bar = 380 μm. Right columns are 3D reconstructions of Z-stack images of DCIS structures (red). One grid unit = 90 μm (arrow in top row, right column, corresponds to the same invasive outgrowth highlighted by arrow in top row, middle column). The inset in the bottom row, middle column, shows an overlay of unlabeled MEPs (DIC) and the absence of outgrowths in DCIS structures (red) when cocultured with MEPs. Scale bar = 100 μm. b MCF10.DCIS cells (DCIS) and WS-12T cells (cancer-associated fibroblasts [CAFs]) were seeded with or without N1ME cells (MEPs) in MAME cultures and imaged live at day 8. Arrowhead points to an invasive outgrowth. Images are 16 contiguous tiled fields (scale bar = 90 μm). Images are representative of at least three independent experiments. Views of 3D reconstructions from a number of angles are shown in Additional file 5: Figure S3