Figure 6
Figure 6. RDTR/SCFHA-displaying LVs allow in vivo gene transfer into immature hCD34+ cells in humanized mice. Newborn Balbc rag2−/−γc−/− mice received sublethal irradiation (2 × 1.5 Gy) and then 2 × 105 hCD34+ CB cells were injected into the fetal liver at day 2 to 4 after birth. After 13 weeks of reconstitution, RDTR/SCFHA-LVs were injected into both femurs of the humanized mice. Two to 3 weeks after injection, the mice were killed and the transduction of the different cell lineages in the BM was analyzed (A-D). The percentage of transduced human cells (hCD45+GFP+) is shown for 3 different injected animals listed in Table 2 (A). In panel B, transduction (GFP+) of immature progenitors (hCD34+), myeloid progenitors (hCD13+), monocytes (hCD14+), and pre- and pro-B cells (hCD19+) in the BM was analyzed by FACS. (C) Transduction in different BM subpopulations by CD34/CD19 double staining. Transduction is shown for very immature, nonlineage-committed CD34+ cells (CD34+CD19−), for pre- and pro-B cells (CD34+CD19+) and further differentiated B cells (CD34−CD19+). In panel D, transduction of immature B cells (CD19+CD20−) and of mature B cells (CD19+CD20+) are presented in the dot blots.

RDTR/SCFHA-displaying LVs allow in vivo gene transfer into immature hCD34+ cells in humanized mice. Newborn Balbc rag2−/−γc−/− mice received sublethal irradiation (2 × 1.5 Gy) and then 2 × 105 hCD34+ CB cells were injected into the fetal liver at day 2 to 4 after birth. After 13 weeks of reconstitution, RDTR/SCFHA-LVs were injected into both femurs of the humanized mice. Two to 3 weeks after injection, the mice were killed and the transduction of the different cell lineages in the BM was analyzed (A-D). The percentage of transduced human cells (hCD45+GFP+) is shown for 3 different injected animals listed in Table 2 (A). In panel B, transduction (GFP+) of immature progenitors (hCD34+), myeloid progenitors (hCD13+), monocytes (hCD14+), and pre- and pro-B cells (hCD19+) in the BM was analyzed by FACS. (C) Transduction in different BM subpopulations by CD34/CD19 double staining. Transduction is shown for very immature, nonlineage-committed CD34+ cells (CD34+CD19), for pre- and pro-B cells (CD34+CD19+) and further differentiated B cells (CD34CD19+). In panel D, transduction of immature B cells (CD19+CD20) and of mature B cells (CD19+CD20+) are presented in the dot blots.

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