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Related Experiment Video

Updated: Jul 13, 2026

Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes

Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes

Published on: December 13, 2012

Analysis of cell differentiation by division tracking cytometry.

Kap-Hyoun Ko¹, Ross Odell, Robert E Nordon

¹Graduate School of Biomedical Engineering, Faculty of Engineering, University of New South Wales, Sydney, Australia.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology

|July 27, 2007

Summary

A new quantitative method using carboxyfluorescein diacetate, succinimidyl ester (CFDA-SE) division tracking analyzes multipotent cell growth and differentiation. This technique reveals distinct proliferation rates between CD34(+) and CD34(-) cord blood cells.

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Simultaneous Assessment of Kinship, Division Number, and Phenotype via Flow Cytometry for Hematopoietic Stem and Progenitor Cells

Simultaneous Assessment of Kinship, Division Number, and Phenotype via Flow Cytometry for Hematopoietic Stem and Progenitor Cells

Published on: March 24, 2023

Area of Science:

Cell Biology
Hematopoiesis
Quantitative Biology

Background:

Multipotent cells, like those in cord blood, are crucial for tissue regeneration and blood formation.
Understanding their growth and differentiation dynamics is key to advancing regenerative medicine and treating blood disorders.
Existing methods may lack the resolution to precisely track cell division and phenotypic changes in heterogeneous cultures.

Purpose of the Study:

To develop and validate a quantitative method for characterizing multipotent cell proliferation and differentiation dynamics.
To apply this method to analyze the behavior of cord blood CD34(+) cells under hematopoietic growth factor stimulation.
To compare the cell cycle kinetics and renewal rates of CD34(+) and CD34(-) cells.

Main Methods:

Utilized time-series carboxyfluorescein diacetate, succinimidyl ester (CFDA-SE) division tracking combined with phenotypic analysis.
Defined population statistics including precursor cell frequency, generation time, and renewal rate.
Employed high-resolution tracking and daily histogram analysis to assign generation numbers and quantify artifacts.

Main Results:

Mitotic activation of CD34(+) cells began within 2 days of cytokine stimulation.
CD34(+) cells exhibited slower cycling (generation time 24.7 h) compared to CD34(-) cells (15.1 h).
A high CD34(+) renewal rate (91%) contributed to a 20-fold increase, while CD34(-) expansion was driven by rapid cycling and differentiation from CD34(+) cells.

Conclusions:

Multitype division tracking offers a detailed quantitative analysis of multipotent cell differentiation.
The method accurately characterizes growth dynamics and reveals differential proliferation rates in heterogeneous cell populations.
This approach provides valuable insights into hematopoietic stem cell behavior and differentiation pathways.