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Efficient Generation of Hematopoietic Precursors and Progenitors From Human Pluripotent Stem Cell Lines.

Woods, Niels-Bjarne LU ; Parker, Aaron S; Moraghebi, Roksana LU ; Lutz, Margaret K; Firth, Amy L; Brennand, Kristen J; Berggren, W Travis; Raya, Angel; Belmonte, Juan Carlos Izpisúa and Gage, Fred H, et al. (2011) In Stem Cells 29(7). p.1158-1164
Abstract
By mimicking embryonic development of the hematopoietic system, we have developed an optimized in vitro differentiation protocol for the generation of precursors of hematopoietic lineages and primitive hematopoietic cells from human embryonic stem cells (ES) and induced pluripotent stem cells (iPS). Factors such as cytokines, extra cellular matrix components, and small molecules, as well as the temporal association and concentration of these factors were tested on seven different human ES and iPS lines. We report the differentiation of up to 84% huCD45+ cells (average 41% ± 16, from 7 pluripotent lines) from the differentiation culture, including significant numbers of primitive CD45+/CD34+ and CD45+/CD34+/CD38- hematopoietic progenitors.... (More)
By mimicking embryonic development of the hematopoietic system, we have developed an optimized in vitro differentiation protocol for the generation of precursors of hematopoietic lineages and primitive hematopoietic cells from human embryonic stem cells (ES) and induced pluripotent stem cells (iPS). Factors such as cytokines, extra cellular matrix components, and small molecules, as well as the temporal association and concentration of these factors were tested on seven different human ES and iPS lines. We report the differentiation of up to 84% huCD45+ cells (average 41% ± 16, from 7 pluripotent lines) from the differentiation culture, including significant numbers of primitive CD45+/CD34+ and CD45+/CD34+/CD38- hematopoietic progenitors. Moreover, the numbers of hematopoietic progenitor cells generated, as measured by colony forming unit assays were comparable to numbers obtained from fresh umbilical cord blood mononuclear cell isolates on a per CD45+ cell basis. Our approach demonstrates highly efficient generation of multipotent hematopoietic progenitors with the highest efficiencies reported to date (CD45+/CD34+) using a single standardized differentiation protocol on several human ES and iPS lines. Our data add to the cumulating evidence for the existence of an in vitro derived precursor to the hematopoietic stem cell (HSC) with limited engrafting ability in transplanted mice, but with multipotent hematopoietic potential. Because this protocol efficiently expands the pre-blood precursors and hematopoietic progenitors, it is ideal for testing novel factors for the generation and expansion of definitive HSCs with long-term repopulating ability. (Less)
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publication status
published
subject
keywords
Differentiation, Hematopoiesis, Hematopoietic progenitors, Pluripotent, stem cells
in
Stem Cells
volume
29
issue
7
pages
1158 - 1164
publisher
AlphaMed Press
external identifiers
  • wos:000292192400014
  • pmid:21544903
  • scopus:79960107459
ISSN
1549-4918
DOI
10.1002/stem.657
language
English
LU publication?
yes
id
6f620a05-5976-4143-8511-d094ae30f871 (old id 1973140)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/21544903?dopt=Abstract
date added to LUP
2011-06-07 14:05:32
date last changed
2017-09-10 03:55:28
@article{6f620a05-5976-4143-8511-d094ae30f871,
  abstract     = {By mimicking embryonic development of the hematopoietic system, we have developed an optimized in vitro differentiation protocol for the generation of precursors of hematopoietic lineages and primitive hematopoietic cells from human embryonic stem cells (ES) and induced pluripotent stem cells (iPS). Factors such as cytokines, extra cellular matrix components, and small molecules, as well as the temporal association and concentration of these factors were tested on seven different human ES and iPS lines. We report the differentiation of up to 84% huCD45+ cells (average 41% ± 16, from 7 pluripotent lines) from the differentiation culture, including significant numbers of primitive CD45+/CD34+ and CD45+/CD34+/CD38- hematopoietic progenitors. Moreover, the numbers of hematopoietic progenitor cells generated, as measured by colony forming unit assays were comparable to numbers obtained from fresh umbilical cord blood mononuclear cell isolates on a per CD45+ cell basis. Our approach demonstrates highly efficient generation of multipotent hematopoietic progenitors with the highest efficiencies reported to date (CD45+/CD34+) using a single standardized differentiation protocol on several human ES and iPS lines. Our data add to the cumulating evidence for the existence of an in vitro derived precursor to the hematopoietic stem cell (HSC) with limited engrafting ability in transplanted mice, but with multipotent hematopoietic potential. Because this protocol efficiently expands the pre-blood precursors and hematopoietic progenitors, it is ideal for testing novel factors for the generation and expansion of definitive HSCs with long-term repopulating ability.},
  author       = {Woods, Niels-Bjarne and Parker, Aaron S and Moraghebi, Roksana and Lutz, Margaret K and Firth, Amy L and Brennand, Kristen J and Berggren, W Travis and Raya, Angel and Belmonte, Juan Carlos Izpisúa and Gage, Fred H and Verma, Inder M},
  issn         = {1549-4918},
  keyword      = {Differentiation,Hematopoiesis,Hematopoietic progenitors,Pluripotent,stem cells},
  language     = {eng},
  number       = {7},
  pages        = {1158--1164},
  publisher    = {AlphaMed Press},
  series       = {Stem Cells},
  title        = {Efficient Generation of Hematopoietic Precursors and Progenitors From Human Pluripotent Stem Cell Lines.},
  url          = {http://dx.doi.org/10.1002/stem.657},
  volume       = {29},
  year         = {2011},
}