Fig. 5.

Myoepithelial cell-conditioned media (MEP-CM) reduce dysplastic phenotype of ductal carcinoma in situ (DCIS) structures formed in mammary architecture and microenvironment engineering (MAME) cultures. DCIS cells were seeded in reconstituted basement membrane overlay cultures containing dye-quenched collagen IV (DQ-collagen IV) in the absence (control) or presence of MEP-CM and imaged live at day 16. Differential interference contrast images are 16 contiguous tiled fields of MCF10.DCIS-lenti-RFP (DCIS; a, top rows) and SUM102-lenti-RFP (SUM102; b, top rows) structures. Scale bars = 180 μm. Fluorescence images are en face views of 3D reconstructions of DCIS (a, bottom rows) and SUM102 (b, bottom rows) structures (red) and their associated DQ-collagen IV degradation products (green). One grid unit = 45 μm. Representative angled views of 3D reconstructions illustrating volume of DCIS (c) and SUM102 (d) structures (red). One grid unit = 180 μm. Volumes of DCIS (e, n = 6) and SUM102 (f, n = 5) structures were quantified in 64 fields (16 contiguous fields per experiment from 4 independent experiments). Data are presented as box-and-whisker plots where the box represents the interquartile range and whiskers represent minimum and maximum values. ** p ≤ 0.007; *** p ≤ 0.0005 as determined by unpaired t test, two-sided. Images are representative of at least three independent experiments. Additional results are shown in Additional file 7: Figure S5, Additional file 8: Video S3, Additional file 9: Video S4, and Additional file 10: Figure S6