Figure 2.

MSCs sustain the expansion and maintenance of GE-HSPCs when used as supportive feeder in standard protocol of gene editing

(A) Schematic representation of the experimental plan used to support UCB-HSPCs undergoing gene editing. BM-MSCs were expanded for 72 h before co-culture with GE-HSPCs. HSPCs were pre-stimulated and recovered upon gene editing on BM-MSC feeder in GE medium 24 h conditioned from BM-MSCs. HSPCs gene edited according to our standard protocol were used as controls. (B and C) Fold change analysis of CD34+ total cell counts (B) and absolute number of HSPCs with a primitive phenotype (C) after 3 days of expansion before editing (3d amp) and after 24 and 72 h recovery after editing in the presence of MSCs compared with standard protocol. (D) Percentage of GFP⁺ GE-HSPCs cells after 72 h expansion on MSC feeder (GE CD34⁺ + MSCs) or in plastic dishes (GE CD34⁺). (E) Absolute number of HDR gene-edited (GFP⁺) GE-HSPCs after co-cultured with MSCs (GE CD34⁺ + MSCs) or expanded in plastic dishes (GE CD34⁺) according to standard protocol. For all the experiments, each error bar shows means ± SEM (n ≥ 3). p values were determined by Mann-Whitney test (∗p ≤ 0.05).