A Continuous Molecular Roadmap to iPSC Reprogramming through Progression Analysis of Single-Cell Mass Cytometry
Identifieur interne : 000200 ( Main/Exploration ); précédent : 000199; suivant : 000201A Continuous Molecular Roadmap to iPSC Reprogramming through Progression Analysis of Single-Cell Mass Cytometry
Auteurs : Eli R. Zunder [États-Unis] ; Ernesto Lujan [États-Unis] ; Yury Goltsev [États-Unis] ; Marius Wernig [États-Unis] ; Garry P. Nolan [États-Unis]Source :
- Cell stem cell [ 1934-5909 ] ; 2015.
Abstract
To analyze cellular reprogramming at the single-cell level, mass cytometry was used to simultaneously measure markers of pluripotency, differentiation, cell-cycle status, and cellular signaling throughout the reprogramming process. Time-resolved progression analysis of the resulting data sets was used to construct a continuous molecular roadmap for three independent reprogramming systems. Although these systems varied substantially in Oct4, Sox2, Klf4, and c-Myc stoichiometry, they presented a common set of reprogramming landmarks. Early in the reprogramming process, Oct4highKlf4high cells transitioned to a CD73highCD104highCD54low partially reprogrammed state. Ki67low cells from this intermediate population reverted to a MEF-like phenotype, but Ki67high cells advanced through the M-E-T and then bifurcated into two distinct populations: an ESC-like NanoghighSox2highCD54high population and a mesendoderm-like NanoglowSox2lowLin28high CD24highPDGFR-αhigh population. The methods developed here for time-resolved, single-cell progression analysis may be used for the study of additional complex and dynamic systems, such as cancer progression and embryonic development.
Url:
DOI: 10.1016/j.stem.2015.01.015
PubMed: 25748935
PubMed Central: 4401090
Affiliations:
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Zunder</name><affiliation wicri:level="2"><nlm:aff id="A1">Department of Microbiology and Immunology, Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, CA 94305, USA</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Microbiology and Immunology, Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, CA 94305</wicri:regionArea><placeName><region type="state">Californie</region></placeName></affiliation></author><author><name sortKey="Lujan, Ernesto" sort="Lujan, Ernesto" uniqKey="Lujan E" first="Ernesto" last="Lujan">Ernesto Lujan</name><affiliation wicri:level="2"><nlm:aff id="A2">Department of Pathology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Pathology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305</wicri:regionArea><placeName><region type="state">Californie</region></placeName></affiliation><affiliation wicri:level="2"><nlm:aff id="A3">Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305</wicri:regionArea><placeName><region type="state">Californie</region></placeName></affiliation></author><author><name sortKey="Goltsev, Yury" sort="Goltsev, Yury" uniqKey="Goltsev Y" first="Yury" last="Goltsev">Yury Goltsev</name><affiliation wicri:level="2"><nlm:aff id="A1">Department of Microbiology and Immunology, Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, CA 94305, USA</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Microbiology and Immunology, Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, CA 94305</wicri:regionArea><placeName><region type="state">Californie</region></placeName></affiliation></author><author><name sortKey="Wernig, Marius" sort="Wernig, Marius" uniqKey="Wernig M" first="Marius" last="Wernig">Marius Wernig</name><affiliation wicri:level="2"><nlm:aff id="A2">Department of Pathology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Pathology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305</wicri:regionArea><placeName><region type="state">Californie</region></placeName></affiliation></author><author><name sortKey="Nolan, Garry P" sort="Nolan, Garry P" uniqKey="Nolan G" first="Garry P." last="Nolan">Garry P. 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Zunder</name><affiliation wicri:level="2"><nlm:aff id="A1">Department of Microbiology and Immunology, Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, CA 94305, USA</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Microbiology and Immunology, Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, CA 94305</wicri:regionArea><placeName><region type="state">Californie</region></placeName></affiliation></author><author><name sortKey="Lujan, Ernesto" sort="Lujan, Ernesto" uniqKey="Lujan E" first="Ernesto" last="Lujan">Ernesto Lujan</name><affiliation wicri:level="2"><nlm:aff id="A2">Department of Pathology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Pathology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305</wicri:regionArea><placeName><region type="state">Californie</region></placeName></affiliation><affiliation wicri:level="2"><nlm:aff id="A3">Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305</wicri:regionArea><placeName><region type="state">Californie</region></placeName></affiliation></author><author><name sortKey="Goltsev, Yury" sort="Goltsev, Yury" uniqKey="Goltsev Y" first="Yury" last="Goltsev">Yury Goltsev</name><affiliation wicri:level="2"><nlm:aff id="A1">Department of Microbiology and Immunology, Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, CA 94305, USA</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Microbiology and Immunology, Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, CA 94305</wicri:regionArea><placeName><region type="state">Californie</region></placeName></affiliation></author><author><name sortKey="Wernig, Marius" sort="Wernig, Marius" uniqKey="Wernig M" first="Marius" last="Wernig">Marius Wernig</name><affiliation wicri:level="2"><nlm:aff id="A2">Department of Pathology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Pathology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305</wicri:regionArea><placeName><region type="state">Californie</region></placeName></affiliation></author><author><name sortKey="Nolan, Garry P" sort="Nolan, Garry P" uniqKey="Nolan G" first="Garry P." last="Nolan">Garry P. Nolan</name><affiliation wicri:level="2"><nlm:aff id="A1">Department of Microbiology and Immunology, Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, CA 94305, USA</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Microbiology and Immunology, Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, CA 94305</wicri:regionArea><placeName><region type="state">Californie</region></placeName></affiliation></author></analytic><series><title level="j">Cell stem cell</title><idno type="ISSN">1934-5909</idno><idno type="eISSN">1875-9777</idno><imprint><date when="2015">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><title>SUMMARY</title><p id="P2">To analyze cellular reprogramming at the single-cell level, mass cytometry was used to simultaneously measure markers of pluripotency, differentiation, cell-cycle status, and cellular signaling throughout the reprogramming process. Time-resolved progression analysis of the resulting data sets was used to construct a continuous molecular roadmap for three independent reprogramming systems. Although these systems varied substantially in Oct4, Sox2, Klf4, and c-Myc stoichiometry, they presented a common set of reprogramming landmarks. Early in the reprogramming process, Oct4<sup>high</sup>Klf4<sup>high</sup> cells transitioned to a CD73<sup>high</sup>CD104<sup>high</sup>CD54<sup>low</sup> partially reprogrammed state. Ki67<sup>low</sup> cells from this intermediate population reverted to a MEF-like phenotype, but Ki67<sup>high</sup> cells advanced through the M-E-T and then bifurcated into two distinct populations: an ESC-like Nanog<sup>high</sup>Sox2<sup>high</sup>CD54<sup>high</sup> population and a mesendoderm-like Nanog<sup>low</sup>Sox2<sup>low</sup>Lin28<sup>high</sup> CD24<sup>high</sup>PDGFR-α<sup>high</sup> population. The methods developed here for time-resolved, single-cell progression analysis may be used for the study of additional complex and dynamic systems, such as cancer progression and embryonic development.</p></div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Californie</li></region></list><tree><country name="États-Unis"><region name="Californie"><name sortKey="Zunder, Eli R" sort="Zunder, Eli R" uniqKey="Zunder E" first="Eli R." last="Zunder">Eli R. Zunder</name></region><name sortKey="Goltsev, Yury" sort="Goltsev, Yury" uniqKey="Goltsev Y" first="Yury" last="Goltsev">Yury Goltsev</name><name sortKey="Lujan, Ernesto" sort="Lujan, Ernesto" uniqKey="Lujan E" first="Ernesto" last="Lujan">Ernesto Lujan</name><name sortKey="Lujan, Ernesto" sort="Lujan, Ernesto" uniqKey="Lujan E" first="Ernesto" last="Lujan">Ernesto Lujan</name><name sortKey="Nolan, Garry P" sort="Nolan, Garry P" uniqKey="Nolan G" first="Garry P." last="Nolan">Garry P. Nolan</name><name sortKey="Wernig, Marius" sort="Wernig, Marius" uniqKey="Wernig M" first="Marius" last="Wernig">Marius Wernig</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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