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Stem Cells Transl Med. 2013 Jun;2(6):420-33. doi: 10.5966/sctm.2012-0139. Epub 2013 May 21.

Lineage-specific purification of neural stem/progenitor cells from differentiated mouse induced pluripotent stem cells.

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Department of Anatomy and Brain Science, Kansai Medical University, Hirakata, Osaka, Japan.


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.


Induced pluripotent stem cells; Neural differentiation; Neural stem cell; Stem/progenitor cell

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