Figure 1. Production of AAV6 via triple transient transfection at low
cell densities. Cells were transfected at 1 or 2 million cells per mL,
and plasmid DNA was delivered on a volumetric basis (VB - 1 µg/mL) or
cell basis (CB - 1 µg/106 cells). A) Cell growth
kinetics and viability. Black arrows indicate the time of transfection,
and dashed arrows indicate the time of viral harvest. B) Transfection
efficiency is described as the percentage of cells expressing the
transgene (GFP) 24 hours post-transfection. C) Volumetric (VG/mL) and
cell-specific viral yields (VG/cell). Ctrl: transfection at 1 ×
106 cells/mL; 2M: at 2 × 106cells/mL. Values represent mean ± standard deviation (n=3). ** P< 0.01, * P < 0.05, by ANOVA followed by
Dunnett’s test.