Comparative AAV-eGFP Transgene Expression Using Vector Serotypes 1–9, 7m8, and 8b in Human Pluripotent Stem Cells, RPEs, and Human and Rat Cortical Neurons

Figure 3

rAAV tropism of in vitro-derived cortical neurons compared to ex vivo-isolated rat cortical neurons. (a) iPSC-derived cortical neurons show dorsal forebrain identity at 82 days in vitro (DIV). The expression of pyramidal neuron markers, Tuj1 (green)/Satb2 (red) or Tuj1 (green)/CTIP2 (red), is shown. Images were captured at 10x magnification. (b) The onset of GFP expression in iPSC-derived cortical neurons (82-86 DIV) at a dosage of 1E6 vg/cell for all rAAV at 48, 72, and 96 h postinfection. Analysis of variance with post hoc pairwise comparisons between time points were performed with statistical significance indicated with , <0.01, and <0.001 after correction for multiple comparisons using Tukey method. (c) The relative AAV-eGFP per cells was observed to be lowest in iPSC-cortical neurons compared to all cells analyzed. The heat map showing relative AAV-GFP expression per cell across all rAAV, dosages, and time in iPSC-derived cortical neurons. The scale bar shows the intensity of AAV-eGFP expression presented as an arbitrary relative fluorescence unit (R.F.U.) per cell. (d) Ex vivo cortical rat neurons were isolated at embryonic day 18. Similar to iPSC-cortical neurons expression, rat cortical neurons express Tuj1 (green)/Satb2 (red) or Tuj1 (green)/CTIP2 (red). Images were captured at 10x magnification. (e) AAV-eGFP expression in rat cortical neurons at a dosage of 1E6 vg/cell for all rAAV at 48, 72, and 96 h. Analysis of variance with post hoc pairwise comparisons between time points were performed with statistical significance indicated with , <0.01, and <0.001 after correction for multiple comparisons using Tukey method. (f) Rat cortical neurons show the highest AAV-GFP expression among cell types, and heat map shows the relative AAV-GFP expression per cell across all rAAV, dosages, and time in rat cortical cells.

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