Figure 5.

coCFTR increases transepithelial anion current in human CF primary airway epithelial cells

CF progenitor basal cells from six donors were transduced with lentiviral vectors HIV-PGK-WTCFTR, HIV-PGK-coCFTR3, or HIV-PGK-GFP at MOI 0.1, 0.25, 0.5, and 1 and seeded on polycarbonate membranes. Cells were allowed to differentiate for 4 weeks in air-liquid interface culture conditions. (A) Epithelia that received GFP were used to estimate the proportion of cells expressing the transgene after differentiation by flow cytometry. (B) CFTR transepithelial chloride and bicarbonate short circuit current was measured in Ussing chambers. Changes (ΔI_SC) in response to (C) F&I and (D) GlyH were quantified. HIV-PGK-coCFTR3 resulted in significantly higher chloride current at MOI 0.1 (∗p < 0.04) and higher bicarbonate current at MOI 0.5 and 1 (∗∗p < 0.02) compared to HIV-PGK-WTCFTR. Points on the graph represent the average of 2−3 epithelia. Each symbol represents cells from an individual donor. Mean ± SE is shown. See also Figure S2.