BaEV-LVs efficiently transduce NSG repopulating hCD34⁺ cells. CB hCD34⁺ cells were prestimulated for 24 hours with a cytokine-cocktail (TPO+SCF+Flk-3L) and transduced the presence of RetroNectin with RDTR-LVs, BaEVTR-LVs, or BaEVRless-LVs at an MOI of 10 for 36 hours. VSV-G-LV transductions were performed at an MOI = 10. The cells were then injected into the liver of newborn NSG mice. On reconstitution for 12 weeks, the different hematopoietic tissues (BM, spleen, and thymus) of these engrafted mice were analyzed for human cell engraftment by anti-hCD45 staining (Table 1). (A) Transduction levels of the different cell lineages in the BM of NSG reconstituted mice. The percentage of GFP⁺ immature early progenitor cells (CD34⁺CD19⁻CD10⁻), pre/pro-B cells (CD34⁺CD19⁺CD10⁻), pro-B cells (CD34⁻CD19⁺CD10⁻), immature and mature B cells (CD34⁻CD19⁺CD10⁺), and myeloid progenitors (CD13⁺) are shown. Data for 2 representative mice from Table 1 for each vector are shown. (B) Transduction levels of different cell lineages in the spleen of NSG reconstituted mice. The percentage of GFP⁺ pre/pro-B cells (CD34⁺CD19⁺CD10⁻), pro-B cells (CD34⁻CD19⁺CD10⁻), immature and mature B cells (CD34⁻CD19⁺CD10⁺), monocytes, and granulocytes (CD14⁺) are shown. Data shown correspond to the mice represented in A. (C) Transduction levels of human thymocyte subpopulations of NSG mice reconstituted with hCD34⁺ cells transduced with BaEVRless-LVs. The percentage of GFP⁺CD4⁺CD8⁺ (DP), CD4⁺CD8⁻ (4-SP), and CD4⁻CD8⁺ (8-SP) is indicated in the upper right quadrant. A representative blot of the BaEVRless mice in Table 1 is shown.