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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.

Author information

1
Department of Anatomy and Brain Science, Kansai Medical University, Hirakata, Osaka, Japan.

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.

KEYWORDS:

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

PMID:
23694811
PMCID:
PMC3673754
DOI:
10.5966/sctm.2012-0139
[Indexed for MEDLINE]
Free PMC Article
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