Early reprogramming regulators identified by prospective isolation and mass cytometry
Identifieur interne : 000971 ( Ncbi/Checkpoint ); précédent : 000970; suivant : 000972Early reprogramming regulators identified by prospective isolation and mass cytometry
Auteurs : Ernesto Lujan [États-Unis] ; Eli R. Zunder [États-Unis] ; Yi Han Ng [États-Unis] ; Isabel N. Goronzy [États-Unis] ; Garry P. Nolan [États-Unis] ; Marius Wernig [États-Unis]Source :
- Nature [ 0028-0836 ] ; 2015.
Abstract
In the context of most induced pluripotent stem (iPS) cell reprogramming methods, heterogeneous populations of nonproductive and staggered productive intermediates arise at different reprogramming time points
Url:
DOI: 10.1038/nature14274
PubMed: 25830878
PubMed Central: 4441548
Affiliations:
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<front><div type="abstract" xml:lang="en"><p id="P2">In the context of most induced pluripotent stem (iPS) cell reprogramming methods, heterogeneous populations of nonproductive and staggered productive intermediates arise at different reprogramming time points<sup><xref rid="R1" ref-type="bibr">1</xref>
–<xref rid="R11" ref-type="bibr">11</xref>
</sup>
. Despite recent reports claiming substantially increased reprogramming efficiencies using genetically modified donor cells<sup><xref rid="R12" ref-type="bibr">12</xref>
,<xref rid="R13" ref-type="bibr">13</xref>
</sup>
prospectively isolating distinct reprogramming intermediates remains an important goal to decipher reprogramming mechanisms. Previous attempts to identify surface markers of intermediate cell populations were based on the assumption that during reprogramming cells progressively lose donor cell identity and gradually acquire iPS cell properties<sup><xref rid="R1" ref-type="bibr">1</xref>
,<xref rid="R2" ref-type="bibr">2</xref>
,<xref rid="R7" ref-type="bibr">7</xref>
,<xref rid="R8" ref-type="bibr">8</xref>
,<xref rid="R10" ref-type="bibr">10</xref>
</sup>
. Here, we report that iPS cell and epithelial markers, such as SSEA1 and EpCAM, respectively, are not predictive of reprogramming during early phases. Instead, in a systematic functional surface marker screen we find that early reprogramming-prone cells express a unique set of surface markers, including CD73, CD49d and CD200 that are absent in fibroblasts and iPS cells. Single cell mass cytometry and prospective isolation show that these distinct intermediates are transient and bridge the gap between donor cell silencing and pluripotency marker acquisition during the early, presumably stochastic reprogramming phase<sup><xref rid="R2" ref-type="bibr">2</xref>
</sup>
. Expression profiling revealed early upregulation of the transcriptional regulators <italic>Nr0b1</italic>
and <italic>Etv5</italic>
in this reprogramming state, preceding activation of key pluripotency regulators such as <italic>Rex1, Dppa2, Nanog</italic>
and <italic>Sox2</italic>
. Both factors are required for the generation of the early intermediate state and fully reprogrammed iPS cells, and thus mark some of the earliest known regulators of iPS cell induction. Our study deconvolutes the first steps in a hierarchical series of events that lead to pluripotency acquisition.</p>
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HfdIndexSelect -h $EXPLOR_AREA/Data/Ncbi/Checkpoint/RBID.i -Sk "pubmed:25830878" \ | HfdSelect -Kh $EXPLOR_AREA/Data/Ncbi/Checkpoint/biblio.hfd \ | NlmPubMed2Wicri -a ThuliumV1
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