Removal of membrane HSPG moderately reduces the percentage of human tracheal epithelial (HTE) cells transduced after rAAV-2 infection. Treated and control HTE cells were infected with AAV2-CMV-EGFP3 for one hour after treated cells had membrane HSPG removed by a 1 hour digestion with 10 mIU of heparinase. Cells were harvested at 48 hours and immediately analyzed for percentage of cells expressing GFP by flow cytometry. Removal of HSPG reduced rAAV-2 transfection efficacy by approximately one third except at MOI of 1000. N = 3 for MOIs 0.1 and 1.0; N = 5 for MOI 10 and 100; N = 8 for MOI = 1000.