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. 2023 Nov 20;32(1):124–139. doi: 10.1016/j.ymthe.2023.11.020

Figure 4.

Figure 4

Delivery of dNs does not affect unstimulated HSPC properties in vitro

(A) Impact of dNs on cell proliferation was assessed in transduced (pool of MOI = 25 and 50) or untransduced quiescent hHSPC. Mean fluorescent intensity (MFI) of the cell proliferation dye was evaluated at the indicated time points by FACS (mean ± SEM; n = 4; circles represent CB-derived HSPC; triangles represent mPB-derived HSPC). (B) p21 mRNA levels were evaluated by qPCR in unstimulated HSPC 48 h post transduction (MOI 50) ± CsH ± dNs (mean ± SEM; n = 3; Dunn’s adjusted Kruskal-Wallis test vs. DMSO = 1). (C) Impact of dNs on apoptosis was assessed in transduced (MOI = 50) or untransduced quiescent hHSPC at 48 h post transduction (mean ± SEM; n = 3–4). (D and E) Impact of dNs on the cell-cycle status of transduced (MOI = 50) or untransduced quiescent hHSPC was evaluated at 24 h (D) and 5 days (E) post transduction. Analysis of the cell-cycle status of the stimulated counterpart at 24 h was included as reference (mean ± SEM; n = 4; Mann Whitney test). (F) The number of total, myeloid, and erythroid CFU was assessed in vitro 2 weeks after cells plating for both quiescent hHSPC exposed or not to dNs and stimulated hHSPC (pool of MOI = 50 and 100) (mean ± SEM; n = 4). Statistical significance is for ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001; ns indicates non-significance.