Figure 2.
Effects of Cell Culture Conditions on JΔNI5 Vector Production
Confluent U2OS-ICP4/27 cells in T75 flasks were infected with JΔNI5 virus at an MOI of 10−5 PFU/cell for 2 h. (A–D) The infected cells were then incubated in media at various pH values (A), temperatures (B), glucose concentrations (C), or serum concentrations (D). Supernatant titers in gc/mL were determined daily by quantitative real-time PCR. Group comparisons were performed with one-way ANOVA with post hoc Dunnett’s multiple comparison test. Statistically significant differences between pH 7.5 and pH 6 are indicated by asterisks (∗p < 0.05). Differences between 33°C and 37°C were statistically highly significant (∗p < 0.05). Statistically significant differences between 4.5 g/L and 0 g/L glucose are indicated (∗p < 0.05). Group A, glucose levels decreasing from the initial 4.5 g/L concentration; group B, glucose levels maintained at ~4.5 g/L; group C, glucose-free media. Data are presented as means ± SD (n = 3).