Multiplexed division tracking dyes for proliferation-based clonal lineage tracing
Details
Publication Year 2018-06-18,Volume 201,Issue #3,Page 1097-1103
Journal Title
Journal of Immunology
Publication Type
Journal Article
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.
Publisher
ASI
Research Division(s)
Immunology
PubMed ID
29914887
NHMRC Grants
NHMRC/1010654NHMRC/1054925
Terms of Use/Rights Notice
Refer to copyright notice on published article.


Creation Date: 2018-06-27 09:13:25
Last Modified: 2018-10-22 01:22:45
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