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Fig. 1 | Stem Cell Research & Therapy

Fig. 1

From: Phenotypic correction of Fanconi anemia cells in the murine bone marrow after carrier cell mediated delivery of lentiviral vector

Fig. 1

Factors affecting 2° TD. a Schematic representation of experimental design. DsRed expressing 293 T cells were used as carrier cells incubated with LV-GFP for 3 h followed by washes. The vector-coated carrier cells are then incubated overnight with 293 T cells in 1:1 ratio. Primary transduced (1° TD) cells will be double-positive for GFP and DsRed and secondary transduced (2° TD) cells will be GFP-positive. b Microscopic imaging of 1° TD and 2° TD HEK293T cells at 48 h post-transduction. c FACS analysis for fluorescent protein expression in HEK293T cell cultures to observe the fate of transduced cells (n = 3). d Long-term analysis of 2° TD and the fate of irradiated carrier cell (shown as Ra) measured by FACS. e Rate of 2° TD is measured after several washes of 1° TD cells with phosphate-buffered saline (PBS). f 2° TD in 293 T cells after transwell migration of murine L1210 cells towards the SDF-1α gradient. L1210-DsRed carrier cells were irradiated and incubated with LV-GFP for 3 h followed by washes. The vector-coated carrier cells are then incubated overnight with 293 T cells in 1:1 ratio. Cells were analyzed for up to 4 weeks. g FACS analysis of primary transduction rate in carrier cells (n = 3; ***p > 0.0001). GFP green fluorescent protein, n/s not significant, SDF stromal-derived factor

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