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. 2019 Feb 26;10:183. doi: 10.3389/fimmu.2019.00183

Figure 1.

Figure 1

Expansion of CB CD34+ cells according to cytokine combinations and differentiation into MDSCs. (A) The scheme of cell expansion or differentiation from CB CD34+ cells. (B) Fold expansion by different cytokine combinations. The culture medium was replaced with each cytokine combinations every week. (C) The FITC conjugated anti-human CD33 were compensated with compensation bead to avoid spillover with PE conjugated anti-human CD11b. (D) Expression of CD11b+CD33 +. The cells were stained with fluorochrome conjugated anti-human CD33 (FITC) and aniti-human CD11b (PE) every week. (E) The CD34+ cells were cultured with human GM-CSF (100 ng/ml) and human SCF (50 ng/ml) for 3 weeks and the cells were stained like (D). The stained cells were sorted by FACS Aria and the sorted cells (CD11b+CD33b+ vs. CD11b−CD33+) were cultured with human GM-CSF (100ng/ml) and human SCF (50ng/ml) for a further 1 week. After one week, the sorted cells were stained like (D) and analyzed by flow cytometry. (F) Expression of HLA-DR, CD14 and CD15. The cells were stained with efluor450-conjugated anti-human HLA-DR, PE-Cy7 anti-human CD14 and APC anti-human CD15 from 3 to 6 weeks, respectively and analyzed by flow cytometry. These experiments were reproduced in 10 individuals (from B to D and F) and 6 individuals (E).