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    Release from cell cycle arrest with Cdk4/6 inhibitors generates highly synchronized cell cycle progression in human cell culture

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    Authors
    Trotter, Eleanor W
    Hagan, Iain M
    Affiliation
    Cell Division Group, CRUK Manchester Institute, The University of Manchester, Alderley Park SK10 4TG, UK.
    Issue Date
    2020
    
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    Abstract
    Each approach used to synchronize cell cycle progression of human cell lines presents a unique set of challenges. Induction synchrony with agents that transiently block progression through key cell cycle stages are popular, but change stoichiometries of cell cycle regulators, invoke compensatory changes in growth rate and, for DNA replication inhibitors, damage DNA. The production, replacement or manipulation of a target molecule must be exceptionally rapid if the interpretation of phenotypes in the cycle under study is to remain independent of impacts upon progression through the preceding cycle. We show how these challenges are avoided by exploiting the ability of the Cdk4/6 inhibitors, palbociclib, ribociclib and abemaciclib to arrest cell cycle progression at the natural control point for cell cycle commitment: the restriction point. After previous work found no change in the coupling of growth and division during recovery from CDK4/6 inhibition, we find high degrees of synchrony in cell cycle progression. Although we validate CDK4/6 induction synchronization with hTERT-RPE-1, A549, THP1 and H1299, it is effective in other lines and avoids the DNA damage that accompanies synchronization by thymidine block/release. Competence to return to cycle after 72 h arrest enables out of cycle target induction/manipulation, without impacting upon preceding cycles.
    Citation
    Trotter EW, Hagan IM. Release from cell cycle arrest with Cdk4/6 inhibitors generates highly synchronized cell cycle progression in human cell culture. Open Biol. 2020;10(10):200200.
    Journal
    Open Biology
    URI
    http://hdl.handle.net/10541/623442
    DOI
    10.1098/rsob.200200
    PubMed ID
    33052073
    Type
    Article
    ae974a485f413a2113503eed53cd6c53
    10.1098/rsob.200200
    Scopus Count
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    All Paterson Institute for Cancer Research

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