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Fig. 2 | Stem Cell Research & Therapy

Fig. 2

From: Cytokine-free directed differentiation of human pluripotent stem cells efficiently produces hemogenic endothelium with lymphoid potential

Fig. 2

Myeloid and NK-lymphoid potential assessment of CHIR99021-induced HE and OP9 coculture-derived progenitors. a Myeloid potential assessment of CHIR99021 derived HE by quantitative colony forming unit (CFU) assay. Error bars are SEM. b Relative proportion of the different types of colonies formed after CHIR99021 induction compared to OP9 coculture induction with and without VEGF. c Phase-contrast images of colony morphology and corresponding cytospin images of E, M, GM, and GEMM colonies formed in semi-solid medium after CHIR99021 induction. d Phase-contrast images of colony morphology and corresponding cytospin images of E, M, GM, and GEMM colonies formed in semi-solid medium after OP9 co-culture induction. e Comparison of CHIR99021-induced and OP9 coculture-induced flow percentages of CD15–, CD56+, and CD56+CD16+ cells after 2 and 4 weeks of culture in lymphoid differentiation medium. The analyzed cells are gated on CD45+ fraction with side scatter characteristics for lymphoid-like cells. Error bars are SEM. f Representative phase-contrast images of cells floating on OP9-DLL4 and representative flow analysis showing CD45+ cells. R1, region of CD45+ cells with side scatter characteristics of lymphoid-like cells. g Representative flow cytometry analysis showing the percentage of CD15– lymphoid fraction of differentiating cells and representative flow analysis showing the expression of CD56+ and CD16+ cells gated on CD15– fraction. E erythroid, M macrophage, G granulocyte component of the GM colony, GM granulocyte/macrophage, GEMM granulocyte, erythroid, monocyte and megakaryocyte colonies

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