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Fig. 2 | Journal of Translational Medicine

Fig. 2

From: Delivery of oncolytic vaccinia virus by matched allogeneic stem cells overcomes critical innate and adaptive immune barriers

Fig. 2

ADSCs promote the oncolysis of resistant tumor cell lines through a combination of virus amplification, tumor cell recruitment and secretion of factors sensitizing the resistant tumor cells to virus infection. a Human ADSC promote the oncolysis of resistant B16 melanoma cells through augmented amplification of the TurboFP635-engineered L14 vaccinia virus. The figure shows fluorescence image analysis of 1 × 106 B16 cells cocultured with 2 × 105 eGFP-labelled RM20 adipose-derived stem cells (×4 magnification) in a 12-well plate. B16 and stem cells were infected together with 1 × 105 pfu virus (MOI = 0.1 to B16) and incubated for up to 72 h. b Plaque assay analysis of vaccinia virus amplification in the coculture experiment as described in a demonstrating that the viral titers recovered from the B16 + ADSC or B16 + A549 cocultures exceed the combined virus output from the individual cells infected in separation. The A549 lung carcinoma cells were used as a highly vaccinia virus permissive positive control. c ADSCs sensitize resistant tumor cells to virus infection. Supernatants from the human RM20 ADSC sensitize B16 melanoma to L14 vaccinia virus infection (×4 magnification). B16 cells were infected in triplicates as in a with L14 vaccinia virus at MOI of 0.1. The effect of supernatants on the infection of B16 cells with the L14 virus was analyzed using the TurboFP635 fluorescence (top panels) and quantitated by plaque assays at 72 h post infection (bottom). Bars represent duplicate measurements ± SD. Statistically significant differences (Student T-test, p < 0.05) based on duplicates or triplicates are marked with asterisks as indicated

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