NextSeq 500 paired end sequencing; IPSC model of ETO2-GLIS2 expression
IPSC model of ETO2-GLIS2 expression
Cell centrifugation Three clones with ETO2-GLIS2 and CTRL iPSC were induced to differentiate into hematological lineage using 2D co-culture system on matrix adapted from Chou S et al. Briefly 3-4 colonies of each iPSC were picked and put into Geltrex (Thermofisher) coated wells in Essential 8 medium. For the first 2 days, the cells are grown in StemPro34 medium supplemented with 1X penicillin/streptomycin (PS, GIBCO), glutamine (2 mM), ascorbic acid (50 μg/ml, Sigma) monothioglycerol (MTG, 15 mM, Sigma), BMP4 (5 ng/mL) and VEGF (50 ng/ml) and CHIR (930 ng/mL). The medium was replenished and supplemented until day 18 with a sequential cytokine cocktail, as shown in Table 2.  Hematopoietic differentiation of iPSC was then assessed at day 13, 15 and 18 using flow cytometry analyses for hematopoietic progenitors (CD34, CD43) and megakaryocytic  markers (CD41, CD42 and CD61). All data were analyzed with FACSCantoII or Fortessa with FACS Diva software (BD Biosciences). Qiagen RNeasy TruSeq Stranded mRNA kit (Illumina) following manufacturers' instructions
Experimental Factor: disease
pediatric leukemia
Experimental Factor: phenotype
CD41+CD42+