Abstract
Embryonic hematopoiesis is a complex process. Elucidating the mechanism regulating hematopoietic differentiation from pluripotent stem cells would allow us to establish a strategy to efficiently generate hematopoietic cells. However, the mechanism governing the generation of hematopoietic progenitors from human embryonic stem cells (hESCs) remains unknown. Here, on the basis of the emergence of CD43+ hematopoietic cells from hemogenic endothelial (HE) cells, we demonstrated that VEGF was essential and sufficient, and that bFGF was synergistic with VEGF to specify the HE cells and the subsequent transition into CD43+ hematopoietic cells. Significantly, we identified TGFβ as a novel signal to regulate hematopoietic development, as the TGFβ inhibitor SB 431542 significantly promoted the transition from HE cells into CD43+ hematopoietic progenitor cells (HPCs) during hESC differentiation. By defining these critical signaling factors during hematopoietic differentiation, we can efficiently generate HPCs from hESCs. Our strategy could offer an in vitro model to study early human hematopoietic development.
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Acknowledgements
We thank Yizhe Zhang, Haisong Liu, Yan Shi, Hongxia Lv, Ting Liu and other colleagues in our laboratory for providing technical assistance and discussions in the preparation of this manuscript. This work was supported by grants from the Ministry of Science and Technology (2011DFA30730), the 111 project, the National Basic Research Program of China (973 program, 2011CB964800 and 2010CB945204) and the Bill & Melinda Gates Foundation (37871 to H Deng). This work was supported by a China Postdoctoral Science Foundation (20100319 to C Wang).
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Supplementary information, Figure S1
(A) (i)Some CD31+ cells with sheet morphology gradually generated semi-adherent hematopoietic cells. (PDF 493 kb)
Supplementary information, Figure S2
(A) Concentration effect of VEGF on CD31+ cells generation. (PDF 58 kb)
Supplementary information, Figure S3
(A) SB 431542 enhances the generation of CD43+ hematopoietic cells from day-4 sorted CD31- cells but does not affect day-6 sorted CD31- cells. (PDF 738 kb)
Supplementary information, Figure S4
(A) H9 cell line was differentiated at the third stage with VEGF and bFGF as control or with VEGF, bFGF and SB 431543 treatment for 2 days. (PDF 367 kb)
Supplementary information, Figure S5
Gene expression analysis of differentiated cells at day 6 with VF and VFSB treatment from D4-D6. (PDF 312 kb)
Supplementary information, Table S1
Signal factors applied in the identification of signals regulating CD31+ HE generationand CD43+ hematopoietic emergence. (PDF 457 kb)
Supplementary information, Data S1
Materials and Methods (PDF 85 kb)
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Wang, C., Tang, X., Sun, X. et al. TGFβ inhibition enhances the generation of hematopoietic progenitors from human ES cell-derived hemogenic endothelial cells using a stepwise strategy. Cell Res 22, 194–207 (2012). https://doi.org/10.1038/cr.2011.138
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DOI: https://doi.org/10.1038/cr.2011.138
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