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    Multiplexed Division Tracking Dyes for Proliferation-Based Clonal Lineage Tracing


    Horton, Miles B. and Prevedello, Giulio and Marchingo, J.M. and Zhou, Jie H.S. and Duffy, Ken R. and Heinzel, Susanne and Hodgkin, Philip D. (2018) Multiplexed Division Tracking Dyes for Proliferation-Based Clonal Lineage Tracing. Journal of Immunology, 201. pp. 1097-1103. ISSN 0022-1767

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    Abstract

    The generation of cellular heterogeneity is an essential feature of immune responses. Understanding the heritability and asymmetry of phenotypic changes throughout this process requires determination of clonal-level contributions to fate selection. Evaluating intraclonal and interclonal heterogeneity and the influence of distinct fate determinants in large numbers of cell lineages, however, is usually laborious, requiring familial tracing and fate mapping. In this study, we introduce a novel, accessible, high-throughput method for measuring familial fate changes with accompanying statistical tools for testing hypotheses. The method combines multiplexing of division tracking dyes with detection of phenotypic markers to reveal clonal lineage properties. We illustrate the method by studying in vitro–activated mouse CD8+ T cell cultures, reporting division and phenotypic changes at the level of families. This approach has broad utility as it is flexible and adaptable to many cell types and to modifications of in vitro, and potentially in vivo, fate monitoring systems.

    Item Type: Article
    Additional Information: This work was supported by the National Health and Medical Research Council of Australia via Project Grant 1010654, Program Grant 1054925, and a fellowship to P.D.H., as well as Australian Government National Health and Medical Research Council Independent Research Institutes Infrastructure Support Scheme Grant 361646. K.R.D. was supported by Science Foundation Ireland Grant 12IP1263. J.M.M. was the recipient of a Sydney Parker Smith Postdoctoral Research Fellowship from the Cancer Council of Victoria. J.H.S.Z. was the recipient of an Australian Postgraduate Award. The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007–2013) under Grant Agreement 317040 (QuanTI)
    Keywords: Multiplexed; Tracking Dyes; Proliferation-Based; Clonal Lineage Tracing;
    Academic Unit: Faculty of Science and Engineering > Research Institutes > Hamilton Institute
    Item ID: 13075
    Identification Number: https://doi.org/10.4049/jimmunol.1800481
    Depositing User: Dr Ken Duffy
    Date Deposited: 19 Jun 2020 15:21
    Journal or Publication Title: Journal of Immunology
    Publisher: American Association of Immunologists
    Refereed: Yes
    Funders: National Health and Medical Research Council of Australia, Science Foundation Ireland (SFI), Cancer Council of Victoria, European Union Seventh Framework Programme
    URI:

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