Figure 6.
Rapamycin treatment on CD34+ cells greatly enhanced engraftment of GFP+ cells in NSG mice spleen and bone marrow at 16 weeks. Cells were prestimulated overnight in SFT6 StemSpan SFEM medium before 24 hours of transduction, and then they were transplanted. (A): Fluorescence-activated cell sorting phenotypic analysis of spleens from NSG mice transplanted with 1 × 106 nontransduced CD34+ cells, transduced CD34+ cells without rapamycin, or transduced CD34+ cells with rapamycin treatment. Upper row: CD45+/GFP+ cells. Lower row: CD4+/GFP+ cells. (B): Statistical analysis of spleens and bone marrows from NSG mice transplanted with 1 × 106 transduced CD34+ cells without rapamycin (n = 10) or with rapamycin treatment (n = 11). The data were analyzed with unpaired two-tailed Student’s t test. ∗∗∗, p < .001; ∗∗∗∗, p < .0001. Abbreviations: GFP, green fluorescent protein; NSG, NOD.Cg-Prkdcscid IL2rgtm1Wjl/SzJ; Rapa, rapamycin; SFEM, serum free expansion media; SFT6, stem cell factor, flt-3 ligand, thrombopoietin, and interleukin-6; Tx, transduction.