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. 2021 Oct 21;16(11):2768–2783. doi: 10.1016/j.stemcr.2021.09.019

Figure 3.

Figure 3

Hematopoietic differentiation potential of iPSC-derived CD34+ cells

(A) Schematic representation of a “spin-EB” protocol to differentiate iPSCs toward hematopoietic stem cells and MKs. Cell culture medium was continuously supplemented with cytokines as indicated. On day 14, suspension cells were harvested for further analysis. Representative cell culture images of indicated days are shown. Scale bars, 50 μm.

(B) Cell number on day 14 of “spin-EB” differentiation calculated for harvested suspension cells/well of 96-well plate. Each data point represents an independent experiment for indicated CALR mutation and genotype shown as mean values ± SD; ∗p > 0.05, ∗∗p > 0.01, n = 5–8 independent differentiation experiments.

(C) Colony-forming unit (CFU) assay of purified iPSC-derived CD34+ cells on day 14 of “spin-EB” differentiation. Red and blue lines refer to significant differences in CFU-E and CFU-M, respectively. Data are shown as mean values ± SD; ∗p > 0.05, ∗∗p > 0.01, ∗∗∗p > 0.001, n = 3 independent experiments per genotype. Morphological appearance of different colony types was assessed by cytospin preparation and Diff-Quik staining. Scale bars, 50 μm.