Tracking differentiating neural progenitors in pluripotent cultures using microRNA-regulated lentiviral vectors.
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.
Department of Experimental Medical Sciences, Wallenberg Neuroscience Center, Lund University, 221 84 Lund, Sweden.
Rohit
R
Sachdeva
Marie E
ME
Jönsson
Jenny
J
Nelander
Agnete
A
Kirkeby
Carolina
C
Guibentif
Bernhard
B
Gentner
Luigi
L
Naldini
Anders
A
Björklund
Malin
M
Parmar
Johan
J
Jakobsson
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019070:Cell Lineage; D002469:Cell Separation; D053595:Embryonic Stem Cells / Q000166:cytology; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D009474:Neurons / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology*
D035683:MicroRNAs; D049452:Green Fluorescent Proteins