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


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.

SUBMITTER: Sachdeva R 

PROVIDER: S-EPMC2895063 | biostudies-literature | 2010 Jun

REPOSITORIES: biostudies-literature

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

Sachdeva Rohit R   Jönsson Marie E ME   Nelander Jenny J   Kirkeby Agnete A   Guibentif Carolina C   Gentner Bernhard B   Naldini Luigi L   Björklund Anders A   Parmar Malin M   Jakobsson Johan J  

Proceedings of the National Academy of Sciences of the United States of America 20100607 25


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 diffe  ...[more]

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