High-Efficiency Transduction of Primary Human Hematopoietic Stem Cells and Erythroid Lineage-Restricted Expression by Optimized AAV6 Serotype Vectors In Vitro and in a Murine Xenograft Model In Vivo
Figure 4
Transcriptional potential of CBAp, HS2-βp and B19p6 promoters in primary human CD34+ and CD36+ human cells following erythroid differentiation.
Approximately 1.5×104 CD34+ cells, and ∼2×104 primary human CD36+ erythroid progenitor cells were cultured with or without Epo (3 U/ml) for various indicated times, and infected with 1×104 vgs/cell scAAV6-Gluc vectors under identical conditions. Transgene expression levels were determined at 18 hrs post-infection at each time-point. Gluc activity at various time-points was normalized to the group without Epo-induction, and the normalized absolute values are shown as average ± standard deviation from triplicates for CD34+ cells (Panel A), and for CD36+ cells (Panel C). Fold changes in transgene expression following erythroid-differentiation were calculated by dividing the normalized Gluc activities by the initial activity on Day 0 (Panels B and D).