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Article: A murine ESC-like state facilitates transgenesis and homologous recombination in human pluripotent stem cells

TitleA murine ESC-like state facilitates transgenesis and homologous recombination in human pluripotent stem cells
Authors
KeywordsAnimal Cell
Article
Cell Culture
Embryonic Stem Cell
Gene
Gene Expression
Gene Targeting
Homologous Recombination
Human
Human Cell
Klf4 Gene
Microarray Analysis
Nanog Gene
Nonhuman
Oct4 Gene
Oncogene C Myc
Pluripotent Stem Cell
Priority Journal
Reverse Transcription Polymerase Chain Reaction
Sox2 Gene
Transgenics
Western Blotting
Issue Date2010
PublisherCell Press. The Journal's web site is located at http://www.cellstemcell.com
Citation
Cell Stem Cell, 2010, v. 6 n. 6, p. 535-546 How to Cite?
AbstractMurine pluripotent stem cells can exist in two functionally distinct states, LIF-dependent embryonic stem cells (ESCs) and bFGF-dependent epiblast stem cells (EpiSCs). However, human pluripotent cells so far seemed to assume only an epiblast-like state. Here we demonstrate that human iPSC reprogramming in the presence of LIF yields human stem cells that display morphological, molecular, and functional properties of murine ESCs. We termed these hLR5 iPSCs because they require the expression of five ectopic reprogramming factors, Oct4, Sox2, Klf4, cMyc, and Nanog, to maintain this more naive state. The cells are "metastable" and upon ectopic factor withdrawal they revert to standard human iPSCs. Finally, we demonstrate that the hLR5 state facilitates gene targeting, and as such provides a powerful tool for the generation of recombinant human pluripotent stem cell lines. © 2010 Elsevier Inc.
Persistent Identifierhttp://hdl.handle.net/10722/141706
ISSN
2015 Impact Factor: 22.387
2015 SCImago Journal Rankings: 13.121
PubMed Central ID
ISI Accession Number ID
Funding AgencyGrant Number
NIH
Dutch Science Organization (NWO)
Gottlieb Daimler- and Karl Benz-Foundation
National Science Council (Taiwan, ROC)
Funding Information:

The authors thank Tim Ahfeldt, Nimet Maherali, and Chad Cowan for providing human iPSCs, Laura Prickett-Rice and Kat Folz-Donahue at the HSCI/MGH Flow Cytometry Core facility cell sorting; and Jason West and Meredith Bryden for technical support. This work was supported by grants from the NIH and a grant from the Dutch Science Organization (NWO). C.B. was supported by the Gottlieb Daimler- and Karl Benz-Foundation. H.-H.C. was supported by the National Science Council (Taiwan, ROC).

References

 

DC FieldValueLanguage
dc.contributor.authorBuecker, Cen_HK
dc.contributor.authorChen, HHen_HK
dc.contributor.authorPolo, JMen_HK
dc.contributor.authorDaheron, Len_HK
dc.contributor.authorBu, Len_HK
dc.contributor.authorBarakat, TSen_HK
dc.contributor.authorOkwieka, Pen_HK
dc.contributor.authorPorter, Aen_HK
dc.contributor.authorGribnau, Jen_HK
dc.contributor.authorHochedlinger, Ken_HK
dc.contributor.authorGeijsen, Nen_HK
dc.date.accessioned2011-09-27T02:58:29Z-
dc.date.available2011-09-27T02:58:29Z-
dc.date.issued2010en_HK
dc.identifier.citationCell Stem Cell, 2010, v. 6 n. 6, p. 535-546en_HK
dc.identifier.issn1934-5909en_HK
dc.identifier.urihttp://hdl.handle.net/10722/141706-
dc.description.abstractMurine pluripotent stem cells can exist in two functionally distinct states, LIF-dependent embryonic stem cells (ESCs) and bFGF-dependent epiblast stem cells (EpiSCs). However, human pluripotent cells so far seemed to assume only an epiblast-like state. Here we demonstrate that human iPSC reprogramming in the presence of LIF yields human stem cells that display morphological, molecular, and functional properties of murine ESCs. We termed these hLR5 iPSCs because they require the expression of five ectopic reprogramming factors, Oct4, Sox2, Klf4, cMyc, and Nanog, to maintain this more naive state. The cells are "metastable" and upon ectopic factor withdrawal they revert to standard human iPSCs. Finally, we demonstrate that the hLR5 state facilitates gene targeting, and as such provides a powerful tool for the generation of recombinant human pluripotent stem cell lines. © 2010 Elsevier Inc.en_HK
dc.languageengen_US
dc.publisherCell Press. The Journal's web site is located at http://www.cellstemcell.comen_HK
dc.relation.ispartofCell Stem Cellen_HK
dc.subjectAnimal Cellen_US
dc.subjectArticleen_US
dc.subjectCell Cultureen_US
dc.subjectEmbryonic Stem Cellen_US
dc.subjectGeneen_US
dc.subjectGene Expressionen_US
dc.subjectGene Targetingen_US
dc.subjectHomologous Recombinationen_US
dc.subjectHumanen_US
dc.subjectHuman Cellen_US
dc.subjectKlf4 Geneen_US
dc.subjectMicroarray Analysisen_US
dc.subjectNanog Geneen_US
dc.subjectNonhumanen_US
dc.subjectOct4 Geneen_US
dc.subjectOncogene C Mycen_US
dc.subjectPluripotent Stem Cellen_US
dc.subjectPriority Journalen_US
dc.subjectReverse Transcription Polymerase Chain Reactionen_US
dc.subjectSox2 Geneen_US
dc.subjectTransgenicsen_US
dc.subjectWestern Blottingen_US
dc.titleA murine ESC-like state facilitates transgenesis and homologous recombination in human pluripotent stem cellsen_HK
dc.typeArticleen_HK
dc.identifier.emailBu, L:leibu@hku.hken_HK
dc.identifier.authorityBu, L=rp01534en_HK
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1016/j.stem.2010.05.003en_HK
dc.identifier.pmid20569691-
dc.identifier.pmcidPMC3162213-
dc.identifier.scopuseid_2-s2.0-77956235887en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-77956235887&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume6en_HK
dc.identifier.issue6en_HK
dc.identifier.spage535en_HK
dc.identifier.epage546en_HK
dc.identifier.eissn1875-9777-
dc.identifier.isiWOS:000278840700014-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridBuecker, C=15724323200en_HK
dc.identifier.scopusauthoridChen, HH=25821966600en_HK
dc.identifier.scopusauthoridPolo, JM=7101663943en_HK
dc.identifier.scopusauthoridDaheron, L=35261444100en_HK
dc.identifier.scopusauthoridBu, L=8510445400en_HK
dc.identifier.scopusauthoridBarakat, TS=35261610200en_HK
dc.identifier.scopusauthoridOkwieka, P=36477613900en_HK
dc.identifier.scopusauthoridPorter, A=7202005281en_HK
dc.identifier.scopusauthoridGribnau, J=6603021763en_HK
dc.identifier.scopusauthoridHochedlinger, K=35600242700en_HK
dc.identifier.scopusauthoridGeijsen, N=6602709223en_HK
dc.identifier.citeulike7359664-

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