RRC ID 44451
Author Maruyama M, Yamashita Y, Kase M, Trifonov S, Sugimoto T.
Title Lineage-specific purification of neural stem/progenitor cells from differentiated mouse induced pluripotent stem cells.
Journal Stem Cells Transl Med
Abstract Since induced pluripotent stem (iPS) cells have differentiation potential into all three germ layer-derived tissues, efficient purification of target cells is required in many fields of iPS research. One useful strategy is isolation of desired cells from differentiated iPS cells by lineage-specific expression of a drug-resistance gene, followed by drug selection. With this strategy, we purified neural stem/progenitor cells (NSCs), a good candidate source for regenerative therapy, from differentiated mouse iPS cells. We constructed a bicistronic expression vector simultaneously expressing blasticidin S resistance gene and DsRed under the control of tandem enhancer of a 257-base pair region of nestin second intron, an NSC-specific enhancer. This construct was efficiently inserted into the iPS genome by piggyBac transposon-mediated gene transfer, and the established subclone was differentiated into NSCs in the presence or absence of blasticidin S. Consequently, incubation with blasticidin S led to purification of NSCs from differentiated iPS cells. Our results suggest that a lineage-specific drug selection strategy is useful for purification of NSCs from differentiated iPS cells and that this strategy can be applied for the purification of other cell types.
Volume 2(6)
Pages 420-33
Published 2013-6-1
DOI 10.5966/sctm.2012-0139
PII sctm.2012-0139
PMID 23694811
PMC PMC3673754
MeSH Animals Cell Differentiation Cell Line Cell Lineage / drug effects Cell Lineage / genetics* Cell Separation / methods* Fetal Proteins / genetics Fetal Proteins / metabolism Gene Expression / drug effects* Genes, Reporter Induced Pluripotent Stem Cells / cytology Induced Pluripotent Stem Cells / metabolism* Intermediate Filament Proteins / genetics* Intermediate Filament Proteins / metabolism Introns Luciferases Mice Microscopy, Fluorescence Nerve Tissue Proteins / genetics* Nerve Tissue Proteins / metabolism Nestin Neural Stem Cells / cytology Neural Stem Cells / drug effects Neural Stem Cells / metabolism* Nucleosides / pharmacology RNA-Binding Proteins / genetics RNA-Binding Proteins / metabolism SOXB1 Transcription Factors / genetics SOXB1 Transcription Factors / metabolism T-Box Domain Proteins / genetics T-Box Domain Proteins / metabolism Transfection
IF 6.429
Times Cited 4
Human and Animal Cells iPS-MEF-Ng-20D-17(APS0001)