Fig. 10. AAV6.2FF transduces the human lung parenchyma.

a Study design to demonstrate the ability of AAV6.2FF to transduce human lung tissue samples generated from precision cut lung slices (PCLS). Metabolic viability (resazurin) assays were conducted on days 2 (D2) and 7 (D7) post tissue harvest. b Resazurin assay results as represented by mean fluorescence with SD demonstrating the metabolic activity of the human PCLS on days 2 and 7. c Bioluminescence detection using IVIS in PCLS from human fetuses (n = 2; 16⁺⁰ and 16⁺³ weeks gestational age) either untreated or transduced with 10⁸ vg per well of AAV-GFP, 10⁸ vg per well of AAV-mCherry, or 10⁵, 10⁸, or 10¹⁰ vg per well of AAV-Luc. Images are 4 days following vector exposure, and the scale is represented as counts. d Quantification of AAV-Luc wells are represented by the mean total flux (in photons per sec) with SD (n = 2 fetuses; with 12 replicates for the AAV-GFP, AAV-mCherry, and No Treatment groups; and 4 replicates for each of the AAV-Luc doses). e Images of human PCLS in a 24-well tissue culture plate 7 days after culture and 3 days after AAV-SPB transduction. An enlarged image of a PCLS in an individual well is indicated by the dashed white box. f Resazurin assay results as represented by mean fluorescence with SD demonstrating viability of the human PCLS 3 days after no treatment (blue), or transduction with 10¹⁰ vg per well of AAV-mSPB (red) or AAV-hSPB (pink). (n = 4 blank; and n = 6 untransduced, n = 6 AAV-mSPB, and n = 6 AAV-hSPB independent tissue slices from a single set of human fetal lungs). The source data for f has been provided as a Source Data file. g Representative confocal images of SP-B (red) and DAPI (blue) of frozen PCLS sections 3 days after transduction with 10¹⁰ vg per well of AAV-mSPB or AAV-hSPB (Top and bottom row image scale bars, 100 μm and 50 μm, respectively).