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Tracking differentiating neural progenitors in pluripotent cultures using microRNA-regulated lentiviral vectors.

Sachdeva, Rohit LU ; Jönsson, Marie LU ; Nelander Wahlestedt, Jenny LU ; Kirkeby, Agnete LU ; Guibentif, Carolina; Gentner, Bernhard; Naldini, Luigi; Björklund, Anders LU ; Parmar, Malin LU and Jakobsson, Johan LU (2010) In Proceedings of the National Academy of Sciences 107(25). p.11602-11607
Abstract
In this study, we have used a microRNA-regulated lentiviral reporter system to visualize and segregate differentiating neuronal cells in pluripotent cultures. Efficient suppression of transgene expression, specifically in undifferentiated pluripotent cells, was achieved by using a lentiviral vector expressing a fluorescent reporter gene regulated by microRNA-292. Using this strategy, it was possible to track progeny from murine ES, human ES cells, and induced pluripotent stem cells as they differentiated toward the neural lineage. In addition, this strategy was successfully used to FACS purify neuronal progenitors for molecular analysis and transplantation. FACS enrichment reduced tumor formation and increased survival of ES cell-derived... (More)
In this study, we have used a microRNA-regulated lentiviral reporter system to visualize and segregate differentiating neuronal cells in pluripotent cultures. Efficient suppression of transgene expression, specifically in undifferentiated pluripotent cells, was achieved by using a lentiviral vector expressing a fluorescent reporter gene regulated by microRNA-292. Using this strategy, it was possible to track progeny from murine ES, human ES cells, and induced pluripotent stem cells as they differentiated toward the neural lineage. In addition, this strategy was successfully used to FACS purify neuronal progenitors for molecular analysis and transplantation. FACS enrichment reduced tumor formation and increased survival of ES cell-derived neuronal progenitors after transplantation. The properties and versatility of the microRNA-regulated vectors allows broad use of these vectors in stem cell applications. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Proceedings of the National Academy of Sciences
volume
107
issue
25
pages
11602 - 11607
publisher
National Acad Sciences
external identifiers
  • wos:000279058000080
  • pmid:20534548
  • scopus:77954923288
ISSN
1091-6490
DOI
10.1073/pnas.1006568107
language
English
LU publication?
yes
id
e43ea878-c86e-4534-9f85-fff6f8bcd25c (old id 1626219)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/20534548?dopt=Abstract
date added to LUP
2010-07-05 19:29:23
date last changed
2018-07-15 04:18:23
@article{e43ea878-c86e-4534-9f85-fff6f8bcd25c,
  abstract     = {In this study, we have used a microRNA-regulated lentiviral reporter system to visualize and segregate differentiating neuronal cells in pluripotent cultures. Efficient suppression of transgene expression, specifically in undifferentiated pluripotent cells, was achieved by using a lentiviral vector expressing a fluorescent reporter gene regulated by microRNA-292. Using this strategy, it was possible to track progeny from murine ES, human ES cells, and induced pluripotent stem cells as they differentiated toward the neural lineage. In addition, this strategy was successfully used to FACS purify neuronal progenitors for molecular analysis and transplantation. FACS enrichment reduced tumor formation and increased survival of ES cell-derived neuronal progenitors after transplantation. The properties and versatility of the microRNA-regulated vectors allows broad use of these vectors in stem cell applications.},
  author       = {Sachdeva, Rohit and Jönsson, Marie and Nelander Wahlestedt, Jenny and Kirkeby, Agnete and Guibentif, Carolina and Gentner, Bernhard and Naldini, Luigi and Björklund, Anders and Parmar, Malin and Jakobsson, Johan},
  issn         = {1091-6490},
  language     = {eng},
  number       = {25},
  pages        = {11602--11607},
  publisher    = {National Acad Sciences},
  series       = {Proceedings of the National Academy of Sciences},
  title        = {Tracking differentiating neural progenitors in pluripotent cultures using microRNA-regulated lentiviral vectors.},
  url          = {http://dx.doi.org/10.1073/pnas.1006568107},
  volume       = {107},
  year         = {2010},
}