Improved Coinfection with Amphotropic Pseudotyped Retroviral Vectors
Figure 3
Coinfection with two retroviral vectors as a function of treatment. Coinfection rates were calculated as cells that are both green and red as a percentage of cells that are either green or red. (a) Mixed green and red vectors (1 : 1) were added to PHF (CRL-2703) in unconcentrated (unc.) form; with spinoculation (sp.; 1200 g, , 2 hours; (1) = with replacement of 50% of viral supernatant after 4 hours; (2) = complete change of medium after 4 hours); with concentration (conc.) from 3 sets of Plat-A plates or 16 sets, using 10,000 g for 3 hours; concentrated using Polybrene flocculation (P); or combined concentration with Polybrene and spinoculation. (b) Mixed green and red vectors (1 : 1) were added to 3T3 cells in unconcentrated (unc.) form; with spinoculation (sp.; 1200 g, , 2 hours; (1) = with replacement of 50% of virus after 4 hours; (2) = change of medium after 4 hours); concentrated using Polybrene flocculation (P); or combined concentration with Polybrene and spinoculation. For both CRL-2703 and 3T3, coinfection rates were calculated 4 days after infection. Averages from three experiments are shown ± standard deviation. Significant differences () are indicated for the combination of spinoculation and Polybrene flocculation versus either procedure alone.
