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. 2013 Dec 3;25(3):212–222. doi: 10.1089/hum.2013.184

FIG. 4.

FIG. 4.

Cell transduction efficiencies of rAAV serotype 1–12 vectors produced in 293 cells compared with Sf9 cells. The transduction efficiencies of rAAV-GFP vectors of various serotypes derived from either 293 or Sf9 cells, respectively, were compared. AAV2 rep/cap-expressing HeLa C12 cells were transduced with 1,000 gp of rAAV-GFP vector of the chosen serotype. Cells were coinfected with adenovirus type 2 at an MOI of 10 (infectious titer). At 48 hr p.i., cells were harvested and the percentage of GFP-positive cells was determined by FACS analysis (GFP+: FL1-H >30). (A) FACS read-outs displaying the transduction efficiencies for rAAV vectors of serotype 2 and produced either in 293 cells or in Sf9 cells. Control cells (designated “no AAV”) were infected with adenovirus only. (B) Comparative analysis of the transduction efficiencies of rAAV vectors with serotype 1–11 capsids derived from freeze–thaw supernatants. The various AAV serotypes display variable absolute transduction efficiencies in C12 cells. To compare transduction efficiencies for each serotype, transduction of rAAVs prepared in 293 cells was set to 1.0. The transduction efficiency of rAAV vectors derived from Sf9 cells from the same serotype is displayed as percentage thereof. The experiments were performed in triplicates and are displayed as mean±standard deviation (n=3). (C) Analysis of rAAV-1, -2, -5, -8, and -12 transduction as described in (B), except that highly purified (AVB-column) rAAV vectors were used. FACS, fluorescence-activated cell sorting.