Figure 6.

PAS positive structures may not necessarily represent the presence of VM: In (A) Micro-CT 3D reconstruction of the SKOV3 ovarian cancer cell line after 4 days culture in Matrigel demonstrating the presence of elevated structures over a cell monlayer. (B) A flatter yet elevated section of the culture did not demonstrate a lumen when reconstructed by Micro-CT, yet this structure stained positive for PAS (lower panel). Size bar represents 100 μm. (C) Reconstruction by Micro-CT demonstrates the presence of a lumen containing structure, as demonstrated by the interior white space. The PAS positive stain is shown in the lower panel. Size bar represents 100 μm. In (D–F) a characterization is shown of structures formed by primary cultures of glioblastoma cells. These samples where obtained with Ethical Committee approval and written patient consent from the Clinical Hospital of the University of Chile, Santiago, Chile. Cell culture was as described previously in Racordon et al. (6). (D) Light microscopy imagery of primary cultured cells grown on matrigel, with an image of the cells grown in plastic in the inlay. Size bar represents 500 μm. (E) Primary cultured glioblastoma cells presented elevated structures over the cell monolayer that stained for PAS. Size bar represents 500 μm. (F) Confocal 3D reconstruction using ZEN 2012 demonstrates that the PAS positive structures observed in (E) are elevated over the cell monolayer but do not possess a lumen.