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. Author manuscript; available in PMC: 2019 Aug 1.
Published in final edited form as: Stem Cell Rev. 2018 Aug;14(4):525–534. doi: 10.1007/s12015-018-9805-1

Figure 4. Generation of hematopoietic cells with ETV2/GATA2 modRNA.

Figure 4

A) Schematic diagram showing in vitro hematopoietic differentiation protocol. Pluripotent stem cells were transfected twice with ETV2/GATA2 combination (arrow). Transfected cells were transferred to collagen coated plate on day 1 in IF9S medium supplemented with hematopoietic cytokines.

B) Representative picture of day 7 hematoendothelial clusters (arrow) from IISH2i-BM9 iPSCs. Floating and adherent cells were analyzed separately with CD34, CD43, CD45, CD144, and CD253a. Dot plot showing live cells gated from the left histogram.

C) Types of hematopoietic colonies formed by floating cells from day 7 post-transfection with ETV2/GATA2 and corresponding Wright-stained cytospin. Granulocytic colonies (G), macrophage colonies (M), and erythroid colonies (E). Scale bar for CFC assay = 200 μm; cytospin = 20 μm.

D) Yields of CD34+43+ cells from human PSCs and E) Yields of colony-forming cells obtained from 2×105 undifferentiated pluripotent stem cells and their CFC potential. Mean±SD from 3 experiments is shown.