Figure 3.

Growth pattern and immunostainings of cancer cell lines on decellularized capillaries. (A) Phase-contrast images show that SH-SY5Y forms loose clusters, ECC-1 forms clusters, and PC3M forms networks. Merged images show the co-localization of the network pattern (formed by cancer cells) with the DC. Clusters appear to localize more randomly by preferring proximity to capillary network. DC is stained with anti- von Willebrand factor (red) or with anti-Collagen IV (green). Cancer cells are stained with anti-α-Actin (green) or anti-ALDH1A1 (red). Imaged with Nikon Eclipse Ti-s inverted fluorescence microscope. (B) A549 co-localize with capillaries, and H460 cells form clusters. Left: the phase contrast image, right: the immunostained image of the culture; vascular basal membrane (Collagen IV, green) and Nuclei (DAPI, blue). Growth pattern of A549 (network) and H460 (cluster) are seen clearly in both images. Cancer cells are easily visualized with DAPI because capillaries no longer contain DNA. Scale bar 100 µm in all images.