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Reactive Oxygen Species Impair the Function of CD90+ Hematopoietic Progenitors Generated from Human Pluripotent Stem Cells

Rönn, Roger E. LU ; Guibentif, Carolina LU ; Saxena, Shobhit LU and Woods, Niels Bjarne LU (2017) In Stem Cells 35(1). p.197-206
Abstract

Cell stressors, such as elevated levels of reactive oxygen species (ROS), adversely affect hematopoietic stem cell (HSC) reconstituting ability. However, the effects of ROS have not been evaluated in the context of hematopoietic development from human pluripotent stem cells (hPSCs). Using our previously described in vitro system for efficient derivation of hematopoietic cells from hPSCs, we show that the vast majority of generated hematopoietic cells display supraphysiological levels of ROS compared to fresh cord blood cells. Elevated ROS resulted in DNA damage of the CD34+ hematopoietic fraction and, following functional assays, reduced colony formation and impaired proliferative capacity. Interestingly, all the... (More)

Cell stressors, such as elevated levels of reactive oxygen species (ROS), adversely affect hematopoietic stem cell (HSC) reconstituting ability. However, the effects of ROS have not been evaluated in the context of hematopoietic development from human pluripotent stem cells (hPSCs). Using our previously described in vitro system for efficient derivation of hematopoietic cells from hPSCs, we show that the vast majority of generated hematopoietic cells display supraphysiological levels of ROS compared to fresh cord blood cells. Elevated ROS resulted in DNA damage of the CD34+ hematopoietic fraction and, following functional assays, reduced colony formation and impaired proliferative capacity. Interestingly, all the proliferative potential of the most primitive hematopoietic cells was limited to a small fraction with low ROS levels. We show that elevation of ROS in hPSC-derived hematopoietic cells is contributed by multiple distinct cellular processes. Furthermore, by targeting these molecular processes with 4 unique factors, we could reduce ROS levels significantly, yielding a 22-fold increase in the most primitive CD90+ CD34+ hematopoietic cells with robust growth capacity. We demonstrate that the ROS reducing factors specifically reduced ROS in more primitive hematopoietic fractions, in contrast to endothelial cells that maintained low ROS levels in the cultures. We conclude that high levels of ROS in in vitro differentiation systems of hPSCs is a major determinant in the lack of ability to generate hematopoietic cells with similar proliferation/differentiation potential to in vivo hematopoietic progenitors, and suggest that elevated ROS is a significant barrier to generating hPSC-derived repopulating HSCs. Stem Cells 2017;35:197–206.

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organization
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type
Contribution to journal
publication status
published
subject
keywords
Functional impairment, Hematopoietic development, Hematopoietic stem cells, Human pluripotent stem cells, Reactive oxygen species
in
Stem Cells
volume
35
issue
1
pages
10 pages
publisher
AlphaMed Press
external identifiers
  • scopus:85000962067
  • wos:000391966700021
ISSN
1066-5099
DOI
10.1002/stem.2503
language
English
LU publication?
yes
id
707c5aa0-f719-480e-b31b-61807221eba8
date added to LUP
2017-01-24 16:20:03
date last changed
2018-11-21 21:29:13
@article{707c5aa0-f719-480e-b31b-61807221eba8,
  abstract     = {<p>Cell stressors, such as elevated levels of reactive oxygen species (ROS), adversely affect hematopoietic stem cell (HSC) reconstituting ability. However, the effects of ROS have not been evaluated in the context of hematopoietic development from human pluripotent stem cells (hPSCs). Using our previously described in vitro system for efficient derivation of hematopoietic cells from hPSCs, we show that the vast majority of generated hematopoietic cells display supraphysiological levels of ROS compared to fresh cord blood cells. Elevated ROS resulted in DNA damage of the CD34<sup>+</sup> hematopoietic fraction and, following functional assays, reduced colony formation and impaired proliferative capacity. Interestingly, all the proliferative potential of the most primitive hematopoietic cells was limited to a small fraction with low ROS levels. We show that elevation of ROS in hPSC-derived hematopoietic cells is contributed by multiple distinct cellular processes. Furthermore, by targeting these molecular processes with 4 unique factors, we could reduce ROS levels significantly, yielding a 22-fold increase in the most primitive CD90<sup>+</sup> CD34<sup>+</sup> hematopoietic cells with robust growth capacity. We demonstrate that the ROS reducing factors specifically reduced ROS in more primitive hematopoietic fractions, in contrast to endothelial cells that maintained low ROS levels in the cultures. We conclude that high levels of ROS in in vitro differentiation systems of hPSCs is a major determinant in the lack of ability to generate hematopoietic cells with similar proliferation/differentiation potential to in vivo hematopoietic progenitors, and suggest that elevated ROS is a significant barrier to generating hPSC-derived repopulating HSCs. Stem Cells 2017;35:197–206.</p>},
  author       = {Rönn, Roger E. and Guibentif, Carolina and Saxena, Shobhit and Woods, Niels Bjarne},
  issn         = {1066-5099},
  keyword      = {Functional impairment,Hematopoietic development,Hematopoietic stem cells,Human pluripotent stem cells,Reactive oxygen species},
  language     = {eng},
  month        = {01},
  number       = {1},
  pages        = {197--206},
  publisher    = {AlphaMed Press},
  series       = {Stem Cells},
  title        = {Reactive Oxygen Species Impair the Function of CD90<sup>+</sup> Hematopoietic Progenitors Generated from Human Pluripotent Stem Cells},
  url          = {http://dx.doi.org/10.1002/stem.2503},
  volume       = {35},
  year         = {2017},
}