Fig. 4.

In vitro glioma tropism and oncovirolytic properties of selected and non-selected human NSC

(a) oHB1.F3 NSC transfected with PDPN or treated with transfection reagent (sham) were subsequently infected with 200 MOI of Ad-Delo3-RGD to induce oncovirolysis of glioma cells or left untreated (0 MOI; mock) for 2 h, followed by a 4 h migration period through 8 µm transwell membranes towards the conditioned medium from LNT-229 glioma cells. Migrated cells were harvested and counted. Number of migrated mock- or Ad-Delo3-RGD infected F3 NSC after 4h of migration was compared between PDPN-transfected vs. untransfected cells by two-tailed t-test (*p<0.05), n=3, each data point represents one biological replicate per group; MOI, multiplicity of infection; Error bars: SEM.

(b) Migrated NSC with and without PDPN overexpression were co-cultured with eGFP-labelled LNT-229 glioma cells. Representative microphotographs from co-cultures taken at the start (day 0), at day 5 and at day 7 of co-culture. Scale bars: 60 µm.

(c) Growth of glioma cells in presence of migrated mock-NSC was quantified as described in methods. Cell density of glioma cells (normalized to the cell density at day 0) in the presence of mock-NSC with and without PDPN transfection was analysed by ANOVA and Bonferroni's multiple comparisons test (*p<0.05), n=3, each data point represents one biological replicate per group out of a single experiment; Error bars: SEM.

(d) Growth of glioma cells in the presence of migrated Ad-Delo3-RGD infected F3 NSC. Cell density of glioma cells at day 5 and day 7 is normalized to that of day 0. For comparison of sham- vs. PDPN-transfected NSC two-tailed t-test (*p<0.05) was used, n=3, each data point represents one biological replicate per group; MOI, multiplicity of infection; Error bars: SEM.