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. Author manuscript; available in PMC: 2015 Mar 1.
Published in final edited form as: Stem Cell Res. 2013 Dec 16;12(2):441–451. doi: 10.1016/j.scr.2013.12.003

Figure 1.

Figure 1

Monolayer differentiation of human ESCs generates multipotent hematopoietic progenitors. (A) Representative flow cytometric analyses of ES cells (SSEA4 vs. SSEA3), day 5 hemogenic mesoderm (CD31 vs. KDR) and day 9 hematopoietic progenitors (CD235 vs. CD41) induced in a serum free/feeder free adherent culture system. (B) Flow cytometric analysis of CD235 vs. CD41 expression on the non-adherent population obtained from days 7, 8 and 9 of the adherent differentiation cultures. (C) QRT-PCR gene expression analyses of ES cells and the day 7, 8, and 9 non-adherent cells shown in B. (D) Day 7, 8 or 9 non-adherent cells shown in B were assayed for progenitor colony forming potential and myeloid, mixed (muti-potential) and erythroid colonies were quantified. (E) Cytospins showing May-Grünwald Giemsa staining of myeloid, erythroid and mixed colonies. Scale bars represent 20 μm. Arrows in the myeloid cytospin indicate cells with neutrophil morphology. (F) Non-adherent cells from H9 and H9-GFP ESC lines at days 7, 8 and 9 of differentiation were mixed at a 1:1 ratio and plated in a methylcellulose colony assay. Mixed colonies were scored based upon GFP expression (GFP+: all cells in a colony GFP positive, GFP−: all cells in a colony GFP negative, GFP+/−: mixture of GFP positive and negative cells in a colony). Scale bars represent 500 μm.