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In vitro human embryonic stem cell hematopoiesis mimics MYB-independent yolk sac hematopoiesis

Stijn Vanhee (UGent) , Katrien De Mulder (UGent) , Yasmine Van Caeneghem (UGent) , Greet Verstichel (UGent) , Nadine Van Roy (UGent) , Björn Menten (UGent) , Imke Velghe (UGent) , Jan Philippé (UGent) , Dominique De Bleser, Bart Lambrecht (UGent) , et al.
(2015) HAEMATOLOGICA. 100(2). p.157-166
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Ghent researchers on unfolded proteins in inflammatory disease (GROUP-ID)
Abstract
Although hematopoietic precursor activity can be generated in vitro from human embryonic stem cells, there is no solid evidence for the appearance of multipotent, self-renewing and transplantable hematopoietic stem cells. This could be due to short half-life of hematopoietic stem cells in culture or, alternatively, human embryonic stem cellinitiated hematopoiesis may be hematopoietic stem cell-independent, similar to yolk sac hematopoiesis, generating multipotent progenitors with limited expansion capacity. Since a MYB was reported to be an excellent marker for hematopoietic stem cell-dependent hematopoiesis, we generated a MYB-eGFP reporter human embryonic stem cell line to study formation of hematopoietic progenitor cells in vitro. We found CD34(+) hemogenic endothelial cells rounding up and developing into CD43(+) hematopoietic cells without expression of MYB-eGFP. MYB-eGFP+ cells appeared relatively late in embryoid body cultures as CD34(+) CD43(+) CD45(-/lo) cells. These MYB-eGFP(+) cells were CD33 positive, proliferated in IL-3 containing media and hematopoietic differentiation was restricted to the granulocytic lineage. In agreement with data obtained on murine Myb(-/-) embryonic stem cells, bright eGFP expression was observed in a subpopulation of cells, during directed myeloid differentiation, which again belonged to the granulocytic lineage. In contrast, CD14(+) macrophage cells were consistently eGFP-and were derived from eGFPprecursors only. In summary, no evidence was obtained for in vitro generation of MYB+ hematopoietic stem cells during embryoid body cultures. The observed MYB expression appeared late in culture and was confined to the granulocytic lineage.
Keywords
C-MYB, DIFFERENTIATION CULTURES, BLOOD-CELLS, PROGENITORS, GENERATION, EXPANSION, ENGRAFTMENT, PRECURSORS, INDUCTION, EMERGENCE

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Citation

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Chicago
Vanhee, Stijn, Katrien De Mulder, Yasmine Van Caeneghem, Greet Verstichel, Nadine Van Roy, Björn Menten, Imke Velghe, et al. 2015. “In Vitro Human Embryonic Stem Cell Hematopoiesis Mimics MYB-independent Yolk Sac Hematopoiesis.” Haematologica 100 (2): 157–166.
APA
Vanhee, S., De Mulder, K., Van Caeneghem, Y., Verstichel, G., Van Roy, N., Menten, B., Velghe, I., et al. (2015). In vitro human embryonic stem cell hematopoiesis mimics MYB-independent yolk sac hematopoiesis. HAEMATOLOGICA, 100(2), 157–166.
Vancouver
1.
Vanhee S, De Mulder K, Van Caeneghem Y, Verstichel G, Van Roy N, Menten B, et al. In vitro human embryonic stem cell hematopoiesis mimics MYB-independent yolk sac hematopoiesis. HAEMATOLOGICA. 2015;100(2):157–66.
MLA
Vanhee, Stijn, Katrien De Mulder, Yasmine Van Caeneghem, et al. “In Vitro Human Embryonic Stem Cell Hematopoiesis Mimics MYB-independent Yolk Sac Hematopoiesis.” HAEMATOLOGICA 100.2 (2015): 157–166. Print.
@article{5986990,
  abstract     = {Although hematopoietic precursor activity can be generated in vitro from human embryonic stem cells, there is no solid evidence for the appearance of multipotent, self-renewing and transplantable hematopoietic stem cells. This could be due to short half-life of hematopoietic stem cells in culture or, alternatively, human embryonic stem cellinitiated hematopoiesis may be hematopoietic stem cell-independent, similar to yolk sac hematopoiesis, generating multipotent progenitors with limited expansion capacity. Since a MYB was reported to be an excellent marker for hematopoietic stem cell-dependent hematopoiesis, we generated a MYB-eGFP reporter human embryonic stem cell line to study formation of hematopoietic progenitor cells in vitro. We found CD34(+) hemogenic endothelial cells rounding up and developing into CD43(+) hematopoietic cells without expression of MYB-eGFP. MYB-eGFP+ cells appeared relatively late in embryoid body cultures as CD34(+) CD43(+) CD45(-/lo) cells. These MYB-eGFP(+) cells were CD33 positive, proliferated in IL-3 containing media and hematopoietic differentiation was restricted to the granulocytic lineage. In agreement with data obtained on murine Myb(-/-) embryonic stem cells, bright eGFP expression was observed in a subpopulation of cells, during directed myeloid differentiation, which again belonged to the granulocytic lineage. In contrast, CD14(+) macrophage cells were consistently eGFP-and were derived from eGFPprecursors only. In summary, no evidence was obtained for in vitro generation of MYB+ hematopoietic stem cells during embryoid body cultures. The observed MYB expression appeared late in culture and was confined to the granulocytic lineage.},
  author       = {Vanhee, Stijn and De Mulder, Katrien and Van Caeneghem, Yasmine and Verstichel, Greet and Van Roy, Nadine and Menten, Bj{\"o}rn and Velghe, Imke and Philipp{\'e}, Jan and De Bleser, Dominique and Lambrecht, Bart and Taghon, Tom and Leclercq, Georges and Kerre, Tessa and Vandekerckhove, Bart},
  issn         = {0390-6078},
  journal      = {HAEMATOLOGICA},
  keyword      = {C-MYB,DIFFERENTIATION CULTURES,BLOOD-CELLS,PROGENITORS,GENERATION,EXPANSION,ENGRAFTMENT,PRECURSORS,INDUCTION,EMERGENCE},
  language     = {eng},
  number       = {2},
  pages        = {157--166},
  title        = {In vitro human embryonic stem cell hematopoiesis mimics MYB-independent yolk sac hematopoiesis},
  url          = {http://dx.doi.org/10.3324/haematol.2014.112144},
  volume       = {100},
  year         = {2015},
}

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