Figure 5.

Optimized gene editing conditions maintain engraftment potential of HSPCs

(A) Percentage of GFP cells within different subpopulations (%GFP in living cells [white bars], %GFP in hCD34⁺ cells [gray bars], and %GFP in hCD34⁺hCD38^-hCD90⁺ [black bars]), 48 h post-transduction, using different MOIs ranging from 1 × 10¹ to 1 × 10⁵ gc/μL of the reporter donor. One to five different biological replicates were performed using 5 independent HD CB-CD34⁺ samples. (B) CFU assay of CB-CD34⁺ cell edited with increased doses of AAVs. Two independent biological replicates from three or four different HD CB-CD34⁺ cells were used. (C) Cell expansion 48 h post-editing relative to the initial number of edited HSPCs. Four independent biological replicates from different HD CB-CD34⁺ cells. (D) GFP percentage in hCD45⁺ cells in the BM of NSG mice transplanted with HSPCs edited with 1 × 10⁴ or 5 × 10⁴ gc/μL GFP-AAV and analyzed 3 mpt. Two independent biological replicates from four different HD CB-CD34⁺ cells were used. Four to seven mice were analyzed in each conditions. Kruskal-Wallis multiple comparison test was performed. ∗∗, p<0.01. ns, not significant.