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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<author><name sortKey="Lujan, Ernesto" sort="Lujan, Ernesto" uniqKey="Lujan E" first="Ernesto" last="Lujan">Ernesto Lujan</name>
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<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>
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<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>
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<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>
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