Multiplexed division tracking dyes for proliferation-based clonal lineage tracing

Miles B. Horton, Giulio Prevedello, Julia M. Marchingo, Jie H.S. Zhou, Ken R. Duffy, Susanne Heinzel, Philip D. Hodgkin

Research output: Contribution to journalArticle

2 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1097-1103
Number of pages7
JournalJournal of Immunology
Volume201
Issue number3
Early online date23 Jul 2018
DOIs
Publication statusPublished - 1 Aug 2018

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    Horton, M. B., Prevedello, G., Marchingo, J. M., Zhou, J. H. S., Duffy, K. R., Heinzel, S., & Hodgkin, P. D. (2018). Multiplexed division tracking dyes for proliferation-based clonal lineage tracing. Journal of Immunology, 201(3), 1097-1103. https://doi.org/10.4049/jimmunol.1800481