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Distinct gene expression signatures in human embryonic stem cells differentiated towards definitive endoderm at single-cell level.

Norrman, Karin; Strömbeck, Anna; Semb, Henrik LU and Ståhlberg, Anders (2012) In Methods
Abstract
Characterization of directed differentiation of pluripotent stem cells towards therapeutically relevant cell types, including pancreatic beta-cells and hepatocytes, depends on molecular markers and assays that resolve the signature of individual cells. Pancreas and liver both have a common origin of anterior definitive endoderm (DE). Here, we differentiated human embryonic stem cells towards DE using three different activin A based treatments. Differentiation efficiencies were evaluated by gene expression profiling over time at cell population level. A panel of key markers was used to study DE formation. Final DE differentiation was also analyzed with immunocytochemistry and single-cell gene expression profiling. We found that cells... (More)
Characterization of directed differentiation of pluripotent stem cells towards therapeutically relevant cell types, including pancreatic beta-cells and hepatocytes, depends on molecular markers and assays that resolve the signature of individual cells. Pancreas and liver both have a common origin of anterior definitive endoderm (DE). Here, we differentiated human embryonic stem cells towards DE using three different activin A based treatments. Differentiation efficiencies were evaluated by gene expression profiling over time at cell population level. A panel of key markers was used to study DE formation. Final DE differentiation was also analyzed with immunocytochemistry and single-cell gene expression profiling. We found that cells treated with activin A in combination with sodium butyrate and B27 serum-free supplement medium generated the most mature DE cells. Cell population studies were useful to monitor the temporal expression of genes involved in primitive streak formation and endoderm formation, while single-cell analysis allowed us to study cell culture heterogeneity and fingerprint individual cells. In addition, single-cell analysis revealed distinct gene expression patterns for the three activin A based protocols applied. Our data provide novel insights in DE gene expression at the cellular level of in vitro differentiated human embryonic stem cells, and illustrate the power of using single-cell gene expression profiling to study differentiation heterogeneity and to characterize cell types and subpopulations. (Less)
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Contribution to journal
publication status
published
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in
Methods
publisher
Elsevier
external identifiers
  • wos:000314555700007
  • pmid:22503774
  • scopus:84872180789
ISSN
1095-9130
DOI
10.1016/j.ymeth.2012.03.030
language
English
LU publication?
yes
id
951e253b-09ca-43db-a5b3-ccd57287cedc (old id 2519427)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/22503774?dopt=Abstract
date added to LUP
2012-05-06 17:47:44
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2017-02-26 04:24:36
@article{951e253b-09ca-43db-a5b3-ccd57287cedc,
  abstract     = {Characterization of directed differentiation of pluripotent stem cells towards therapeutically relevant cell types, including pancreatic beta-cells and hepatocytes, depends on molecular markers and assays that resolve the signature of individual cells. Pancreas and liver both have a common origin of anterior definitive endoderm (DE). Here, we differentiated human embryonic stem cells towards DE using three different activin A based treatments. Differentiation efficiencies were evaluated by gene expression profiling over time at cell population level. A panel of key markers was used to study DE formation. Final DE differentiation was also analyzed with immunocytochemistry and single-cell gene expression profiling. We found that cells treated with activin A in combination with sodium butyrate and B27 serum-free supplement medium generated the most mature DE cells. Cell population studies were useful to monitor the temporal expression of genes involved in primitive streak formation and endoderm formation, while single-cell analysis allowed us to study cell culture heterogeneity and fingerprint individual cells. In addition, single-cell analysis revealed distinct gene expression patterns for the three activin A based protocols applied. Our data provide novel insights in DE gene expression at the cellular level of in vitro differentiated human embryonic stem cells, and illustrate the power of using single-cell gene expression profiling to study differentiation heterogeneity and to characterize cell types and subpopulations.},
  author       = {Norrman, Karin and Strömbeck, Anna and Semb, Henrik and Ståhlberg, Anders},
  issn         = {1095-9130},
  language     = {eng},
  month        = {04},
  publisher    = {Elsevier},
  series       = {Methods},
  title        = {Distinct gene expression signatures in human embryonic stem cells differentiated towards definitive endoderm at single-cell level.},
  url          = {http://dx.doi.org/10.1016/j.ymeth.2012.03.030},
  year         = {2012},
}