Article Title: The Oncolytic Virus VSV-GP Is Effective against Malignant Melanoma
Figure Lengend Snippet: Melanoma cell lines and primary tumor cultures were efficiently infected and killed by VSV-GP. ( A ) The tropisms of VSV-GP and VSV wild-type for several human (A375, MDA-mB-435, MJS, NW1539, SK-MEL3, SK-MEL5) melanoma cell lines, one mouse (B16-OVA) melanoma cell line, and one dog (UCDK9-M1) melanoma cell line were analyzed. Cells were seeded as monolayers, and infected with 10-fold serial dilutions of the single cycle infective VSV*MQΔG-GP or VSV*MQΔG-G virus. BHK-21 cells were used as a reference. Sixteen hours post-infection, cells were analyzed for the percentage of GFP-positive cells via flow cytometry, and the titer for both viruses on each cell line was determined. The titers are given relative to the titer on BHK-21. Bars represent the means ± SEM of one representative of at least two independent experiments using duplicate or triplicate samples. For the control cell line, BHK-21, the mean and SEM of one representative experiment is shown; ( B ) monolayers of melanoma cell lines were infected with an MOI (multiplicity of infection) of 0.1 of VSV wild-type or VSV-GP in dodecaplicates. After 24 or 48 h, the viability of cells was determined using WST-1 assay. Values were normalized to the mock-infected sample, and represented as a percentage of surviving cells. Bars represent the mean ± SEM of one representative experiment of at least three independent experiments; ( C , D ) short-term cultures of human melanoma cells were seeded in 24-well plates, and 36 hours afterwards were infected with an MOI of 0.1 of VSV-GFP or VSV-GP-GFP. As control, normal melanocytes were used. Twenty-four hours post-infection, cells were analyzed in the fluorescence microscope for GFP positive, i.e., virus infected, ( C ) and living cells ( D ). Ten microscopic fields were assessed per condition. Bars represent mean ± SEM.
Article Snippet: SK-MEL3 (from DSMZ) were maintained in McCoy’s A5 medium (Gibco) supplemented with 10% FCS, 100 U/mL penicillin, and 100 mg/mL Streptomycin.
Techniques: Infection, Multiple Displacement Amplification, Flow Cytometry, Cytometry, WST-1 Assay, Fluorescence, Microscopy