Figure 2

Lumen–containing tubular structures are present in ovarian cancer cell lines in culture. (a) 3D-reconstruction of X-ray microtomography: panel a, Reconstructed view of the landscape of a 4 day 3D-culture of SKOV3 cells. Elevated structures with tubular-like appearances are clearly visible. The structures within the white rectangle are shown in higher magnification in panel b and c, with the arrowhead denoting a seemingly tubular structure projecting above the flat cell aggregates. A cross-section of this structure is shown to demonstrate an air-filled space (panel a,c) with an estimated diameter of 50 μm. (b) 4 day-old 3D-culture of GFP-HEY cells as demonstrated in clear field (a) and fluorescence (b). (c) Confocal 3D reconstruction of 4 day-old 3D-culture of GFP-Hey cells with the color map discriminating between the planes (height of structures) as indicated in the key. (b) Amplification of region A1 demonstrating the presence of a cell-containing structure in a higher plane respect to the monolayer of cells. When rotated, same image reveals the presence of a lumen (c). (d) A confocal microscopy Z-stack reconstruction of region A1 demonstrating the presence of a cell containing tubular structure. Z-Stack demonstrates a continuous upper monolayer (1), with central walled structures with a hollow center (2) and a continuous lower monolayer (3). This is also shown clearly in the computer-generated cross-section in panel b. (e). Imaris software reconstruction of area A2 from panel c, showing cancer cells in two distinct planes, the red plane being elevated in respect to the lower green plane. A 90° backward rotation of this structure clearly shows the presence of a lumen (b). This representation, that exclusively shows cancer cell fluorescence, demonstrates a fenestrated appearance. Please also refer to Supplementary Video 2.