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Updated: Aug 23, 2025

Measuring Cell Cycle Progression Kinetics with Metabolic Labeling and Flow Cytometry
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CycleFlow simultaneously quantifies cell-cycle phase lengths and quiescence in vivo.

Adrien Jolly1, Ann-Kathrin Fanti2, Csilla Kongsaysak-Lengyel2

  • 1Division of Theoretical Systems Biology, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany.

Cell Reports Methods
|October 31, 2022
PubMed
Summary

CycleFlow quantifies cell proliferation and quiescence using a novel mathematical model. This method reveals significant cell cycle heterogeneity in T cell progenitors, improving quantitative cell cycle analysis.

Keywords:
BrdU labelingEdU labelingG0cell cyclecell proliferationcell-cycle arrestnon-Markovian modelquiescencestatistical inferencethymocyte development

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Area of Science:

  • Cell Biology
  • Quantitative Biology
  • Immunology

Background:

  • Cell populations exhibit proliferative heterogeneity, with both proliferating and quiescent cells.
  • Accurate quantification of quiescent cells and cell cycle progression remains challenging.

Purpose of the Study:

  • To develop a robust method for inferring cell cycle dynamics, including quiescence.
  • To quantify proliferative heterogeneity in cell populations.

Main Methods:

  • Developed CycleFlow, a method based on a mathematical cell cycle model.
  • Utilizes standard pulse-chase experiments with thymidine analogs.
  • Incorporates realistic waiting time distributions for G1, S, G2/M, and G0 phases.

Main Results:

  • CycleFlow successfully quantifies quiescent subpopulations and cell cycle progression.
  • Validated in vitro with an exponentially growing cancer cell line.
  • Revealed significant proliferative heterogeneity in vivo T cell progenitors, with rapid cycling and subsequent quiescence observed in a subset of CD4+CD8+ cells.

Conclusions:

  • CycleFlow provides a robust and comprehensive method for quantitative cell cycle analysis.
  • The method is suitable for routine use in biological research.
  • Uncovered previously uncharacterized proliferative heterogeneity in T cell progenitors.