Figure 5

CD34⁺PRLR⁺ progenitors enhance the generation of CD56⁺ lymphocytes from CD34⁺PRLR⁻ progenitors. UCB-derived CD34⁺ HSCs were expanded for four days using FLT3L, LDL, SCF and TPO. Cells were then differentiated with or without PRL and SR1 for 21 days. The CD34⁺PRLR⁺ and CD34⁺PRLR⁻ cells were sorted at day 4 by FACS followed by differentiation for 21 days. CD34⁺PRLR⁻ and CD34⁺PRLR⁺ co-culture was done in a 2:1 ratios. (A–C) CD56 staining by CD34⁺PRLR⁻, CD34⁺PRLR⁻ + CD34⁺PRLR⁺ co-culture and CD34⁺PRLR⁻ + CD34⁺PRLR⁺ co-culture in transwell system at day 21. Representative histograms (A) percentage in bar graphs (B) and absolute number in bar graphs (C) of CD56⁺ cells are shown (n = 3). (D,E) Cells differentiating in the presence or absence of PRL and SR1 were stained for CD56 at day 21 of culture and the percentage (D) and absolute number (E) of CD56⁺ cells are shown in bar graphs (n = 7). (F) Surface expression of CD56, CD94, CD127 and CD336 by cells differentiating in the presence or absence of PRL and SR1 at day 21. Representative histograms and values represent the percentage (n = 7). (B–E) Data are shown as means ± SD, One-way ANOVA and significance is shown (* = p < 0.01; ** = p < 0.001; **** = p < 0.0001).