Effect of vector titer on transduction efficiency and cell viability (measured by flow cytometry two days after transduction).

<p>Means and standard deviations of three independent experiments are shown. (A) Transduced = GFP-positive cells. Transduction with scAAV2 resulted in significantly higher GFP expression rates compared to ssAAV2 (** = p<0.01, *** = p<0.001). (B) Dead = 7-AAD-positive cells. There were no significant differences between cells transduced with sc- and ssAAV2 or between transduced cells and negative controls (one-way ANOVA, p = 0.062). (C) Apoptotic = PO-PRO-1-positive cells. There were no significant differences between cells transduced with sc- and ssAAV2 or between transduced cells and negative controls (one-way ANOVA, p = 0.578).</p

Confocal microscopy images of endothelial cells of human donor corneas six days after transduction.

<p>(A) transduced with 100,000 GOI ssAAV2; (B) transduced with 100,000 GOI scAAV2. Six random microscopic fields per corneal piece were evaluated. For each vector, one exemplary microscopic field is shown (blue = DAPI, green = GFP, pink = Alexa 647, red = TUNEL).</p

Metabolic activity of transduced cells as a sign of cell viability (analyzed by MTT-assay two days after transduction).

<p>Means and standard deviations of three independent experiments are shown. There was no significant difference between transduced cells and negative controls (one-way ANOVA; p = 0.415).</p

Confocal microscopy images of HCEC-12 cells transduced with different vector titers two days after transduction.

<p>(A) cells transduced with ssAAV2; (B) cells transduced with scAAV2.</p

GFP expression of HCEC-12 cells five days after transduction (flow cytometry).

<p>Means and standard deviations of three independent experiments are shown. GFP expression after transduction with scAAV was significantly higher compared to ssAAV2 (* = p<0.05, ** = p<0.01).</p

GFP-expression levels of transduced HCEC-12 cells over time (flow cytometry).

<p>Means and standard deviations of three independent experiments are shown. (A) vector titers 1,000-10,000 GOI; (B) vector titers 25,000–100,000 GOI.</p