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. 2019 Nov 28;105(8):2044–2055. doi: 10.3324/haematol.2019.226936

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Figure 1. — Low doses (LD) of ionizing radiations (IR) exposure of human hematopoietic stem progenitor cells (HSPC) leads to deficient serial colony forming unit-cell assay (CFU-C) and primary and extended long-term culture initiating cell (LTC-IC) potentials. CD34⁺ CD38^low CD45RA⁻ CD90⁺ HSPC were sorted from pools of independent cord blood (CB) samples by cell sorting and exposed to the indicated IR doses prior to in vitro cultures. (A) LTC-IC assay in limiting dilution (pool of 2 experiments, 120 wells/IR dose). Irradiated CD34⁺ CD38^low CD45RA⁻CD90⁺ HSPC were seeded on MS5 stromal cells in limiting dilution for five weeks then plated in methylcellulose for 12 days. LTC-IC frequency was calculated using LCALC software. (B) Primary CFU-C assay (cumulative results from 4 independent experiments with HSPC isolated from 4 independent pools of CB samples). HSPC (500 cells/plate) were plated in CFU-C condition for 12-14 days and the number (nb) of CFU-C was quantified. Results are normalized to the non-irradiated conditions. (C) Primary CFU-C were pooled and replated in methylcellulose for 12-14 days. Shown are the nb of secondary CFU-C. Results are normalized to the sham-irradiated conditions (cumulative results from 3 independent experiments). Results are shown as mean±standard error of mean. ^**P<0.01, ^***P<0.001, ^****P<0.0001 (Mann-Whitney statistics).