The Comparative Utility of Viromer RED and Lipofectamine for Transient Gene Introduction into Glial Cells

Figure 3

(a) Viromer RED shows differential viability in cell lines and primary cells: HEK293, primary rat astrocytes, primary rat Schwann cells, and BV2 microglial cell were treated with transfection reagents with or without cargo (blue = untreated control, green = Lipofectamine 3000 with enhancer, gray = Viromer RED, purple = Lipofectamine 3000 with enhancer with pGL4.13 luciferase plasmid, orange = Viromer RED with pGL4.13 luciferase plasmid, and red = 1% hydrogen peroxide (H₂O₂) treated cells for 5 min before the addition of CellTiter-Fluor reagent). Cells were assessed for viability 24 hr after transfection using CellTiter-Fluor reagent. Absolute fluorescence (RFU measured at 380 nm Excitation/505 nm Emission) was normalized to mean fluorescence of nontransfected cells and fold change reported. Comparisons were made within the untreated control (), within the transfection reagents (with or without cargo), (&) and across the transfection reagents (within cargo or no cargo group) (#) in the respective cells/cell lines. Values represent mean ± 1 SD. ; ; . (b) Viromer RED transfected cells show a higher luminescence as compared to those transfected with Lipofectamine 3000: HEK293, primary rat astrocytes, primary rat Schwann cells, and BV2 microglial cell lines were treated with transfection reagents, with or without cargo (blue = untreated control, green = Lipofectamine 3000 with enhancer, gray = Viromer RED, purple = Lipofectamine 3000 with enhancer with pGL4.13 luciferase plasmid, and orange = Viromer RED with pGL4.13 luciferase plasmid). Cells were lysed 24 hr after transfection following the assessment of viability (absolute fluorescence (RFU)) using CellTiter-Fluor reagent. Luminescence (RLU) was measured using the luciferase assay reagent. The RLU/RFU ratio data was normalized to the mean RLU/RFU ratio of the nontransfected cells. Comparisons were made across the transfection reagents (with cargo group) (#) in the respective cells/cell lines. Values represent mean ± 1 SD. .

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