title: Lineage-specific purification of neural stem/progenitor cells from differentiated mouse induced pluripotent stem cells.
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.
authors.affiliation: Department of Anatomy and Brain Science, Kansai Medical University, Hirakata, Osaka, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Maruyama
authors.forename: Yuji
authors.initials: Y
authors.lastname: Yamashita
authors.forename: Masahiko
authors.initials: M
authors.lastname: Kase
authors.forename: Stefan
authors.initials: S
authors.lastname: Trifonov
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Sugimoto
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage / Q000187:drug effects / Q000235:genetics*; D002469:Cell Separation / Q000379:methods*; D005326:Fetal Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects*; D017930:Genes, Reporter; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D007381:Intermediate Filament Proteins / Q000235:genetics* / Q000378:metabolism; D007438:Introns; D008156:Luciferases; D051379:Mice; D008856:Microscopy, Fluorescence; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism; D064231:Nestin; D058953:Neural Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009705:Nucleosides / Q000494:pharmacology; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D020825:T-Box Domain Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D000098908:Brachyury Protein
chemical_list: D005326:Fetal Proteins; D007381:Intermediate Filament Proteins; D008156:Luciferases; D009419:Nerve Tissue Proteins; D064231:Nestin; D009705:Nucleosides; D016601:RNA-Binding Proteins; D055748:SOXB1 Transcription Factors; D020825:T-Box Domain Proteins; D000098908:Brachyury Protein; C104786:Msi1h protein, mouse; C577485:Nes protein, mouse; C527783:Sox2 protein, mouse; C004500:blasticidin S
keywords: Induced pluripotent stem cells; Neural differentiation; Neural stem cell; Stem/progenitor cell