Fig. 3.

3D astrocyte scaffold invasion assay. a Assay design. b Confocal xy-sections of astrocyte culture (3 days) stained for F-actin, laminin and collagen type IV (Col IV). c 3D reconstruction (confocal z-stack, 90 μm, horizontal and orthogonal projections) of E-98 and U-251 cell invasion from spheroids (S) into 3-day old mouse astrocyte scaffolds. Glioma cells were identified by vimentin staining with human-specific antibody and constitutive expression of H2BeGFP in the nucleus, and murine astrocytes using phalloidin (F-actin). Arrowheads point to contacts between glioma cells via dendrite-like filaments. Asterisk, detached single cell. d Average migration distance of U-251 and E-98 cells invading astrocyte scaffolds matured for 2, 3 or 10 days before addition of glioma spheroids. Values display median (black line), 25/75 percentiles (boxes) and maximum/minimum (whiskers) from three independent experiments. Scale bars 50 μm