Thymic repopulation by cultured CD34⁺CD38⁻Lin⁻ cells in vivo. Flow cytometric analysis of SCID-hu thymocytes, 30 days after injection with transduced CD34⁺CD38⁻Lin⁻ cells, precultured for 3 weeks in TPO + FL + SCF. (A) Dot plot shows anti-HLA–A3 FITC staining plus GFP fluorescence versus CD3 APC staining. (B) Dot plot shows GFP fluorescence versus CD3 APC staining; cells were not stained with anti–HLA-A3 FITC. (C) Dot plots show CD4 APC versus CD8β and CD4 APC versus CD3 PE staining for cells stained with anti–HLA-A3 FITC as in (A), gated on either FITC⁻GFP⁻ cells (acceptor cells) or FITC⁺GFP⁺ cells (donor cells); and for cells not stained with anti-HLA–A3 FITC as in (B), gated on GFP⁺ cells (transduced GFP⁺ donor cells). Quadrants were set to include 99% of cells stained with isotypic control and GFP⁻ cells in lower left quadrants. Data shown are representative of 2 independent experiments with 2 different donors.