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Intraclonal heterogeneity in CMML
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Figure 4. Functional heterogeneity of the patient’s induced pluripotent stem cell-derived hematopoietic cells. (A) Control and chronic myelomonocytic leukemia (CMML) induced pluripotent stem cell (iPSC)-derived CD34+CD43+ cells were cultured in liquid medium for 10 days in the presence of 50 ng/mL stem cell factor (SCF), 10 ng/mL Fms-like tyrosine kinase 3 ligand (FLT3L), 10 ng/mL interleukin-3 (IL-3), 10 ng/mL interleukin-6 (IL-6), 50 ng/mL thrombopoietin (TPO), 1 U/mL ery- thropoietin (EPO), 10 ng/mL granulocyte-macrophage colony-stimulating factor (GM-CSF), 10 ng/mL granulocyte colony-stimulating factor (G-CSF), and 10 ng/mL monocyte colony-stimulating factor (M-CSF). (B) Total number of hematopoietic cells generated by 5,000 CD34+CD43+ cells cultured in liquid medium for 10 days. (C). Fractions of viable, DAPI-negative cells measured on day 10; Kruskal-Wallis test. (D) Representative flow cytometry analysis of CD33+CD14+ cells generated in liquid culture by the Co3 and A2 clones. (E-I) Fractions of CD33+CD14+ cells (E and insert), CD33+CD14-CD41- cells (F), CD123+CD33-CD235a-CD14-CD41- cells (G), CD235a+CD14-CD41- cells (H), and CD41+CD14- cells (I) generated in liquid culture by the indicated clones; Kruskal-Wallis test. (J) Radar representation of the dif- ferentiation potential of CD34+CD43+ hematopoietic cells derived from control iPSC (green), KRAS wildtype CMML iPSC (blue) and KRAS(G12D) CMML iPSC (red). Bars: mean ± standard deviation. *P<0.05; ***P<0,001 ****P<0.0001
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