Figure 2.

Cell-to-Cell Contacts Are Important to Improve Lentiviral Transduction in High-Density Human CD34⁺ Cell Culture

(A) We transduced half of the CD34⁺ cells with a GFP-expressing lentiviral vector at MOI 50, whereas the other half of CD34⁺ cells were stained with a far-red fluorescent dye. One hour later, transduced CD34⁺ cells were washed with PBS medium and co-cultured (1:1 ratio) with far-red stained CD34⁺ cells (non-transduced) among various cell densities (5e4/mL, 1e5/mL, 2e5/mL, 5e5/mL, 1e6/mL, 2e6/mL, and 4e6/mL). (B) We evaluated %GFP in far-red positive and -negative cells 3 days after transduction. (C) In addition, we evaluated cell proliferation of human CD34⁺ cells cultured for 7 days at various cell densities using cell proliferation dye. Values: mean ± SE. Experiments were performed in duplicate (B) or triplicate (C).