Transduction of human and macaque CD34⁺ cells.

CD34⁺ cells derived from human mobilized blood (A) and from cynomolgus macaque bone marrow (B) were prestimulated by overnight incubation with TPO and were transduced for 16 hours at different MOIs with SIV vectors pseudotyped with VSV-G (triangles), MLV-A GP (closed circles), GALV/TR GP (open circles), or RD114/TR GP (closed squares). For each sample of CD34⁺ cells, transductions were performed in duplicate: in the absence or in the presence of CH-296 retronectin polypeptides coated on the plates. After infection, cells were washed in PBS and cultured in the presence of Flt3-L, TPO, and SCF for an additional 3 days until transduction efficiency was assessed. The dose-response curves of representative experiments are shown for the same batches of CD34⁺ cells as well as the statistical analyses of the maximal transduction efficiencies of at least 4 experiments performed with CD34⁺ cells derived from different donors and stocks of pseudotyped vectors.