Vol 51, No 3 (2013)
Original paper
Submitted: 2013-07-23
Accepted: 2013-09-23
Published online: 2013-11-07
Get Citation

Monitoring cell proliferation in vitro with different cellular fluorescent dyes

Joanna Zolnierowicz, Magdalena Ambrozek-Latecka, Jerzy Kawiak, Danuta Wasilewska, Grazyna Hoser
DOI: 10.5603/FHC.2013.0027
·
Folia Histochem Cytobiol 2013;51(3):193-200.
Vol 51, No 3 (2013)
ORIGINAL PAPERS
Submitted: 2013-07-23
Accepted: 2013-09-23
Published online: 2013-11-07

Abstract

There are few methods for quantifying cell proliferation. Those tests describe the proliferation kinetics of a cell population, but they do not report the history of single cells, the number and frequency of cell divisions, or the precursor cell frequency. Cell-tracking assays based on dilution of the green fluorescent protein labelling dye, CFSE, has become the standard for monitoring cell proliferation. Other labelling dyes, e.g. CellTrace Violet and CellVue Claret, are also used for the same purpose. This study aimed to compare these three cell labelling methods for analysing the kinetics of cell viability, proliferation, and precursor cell frequency. Human peripheral blood mononuclear cells stimulated with Concanavalin A (ConA) were used as a model system. After labelling with a cell-tracking dye cells were divided into groups with and without ConA stimulation. From the 5th to 8th day, cells were collected and analysed with flow cytometry. Cell viability was not significantly different between labelled and unlabelled cells that received ConA stimulation. The proliferative fraction, proliferation index, and nonproliferative fraction were not significantly different among lymphocytes labelled with different dyes. Precursor cell frequency was also similar among cells labelled with the three cell-tracing dyes. The practical conclusion from our observations is that the results from cells labelled with different tracers may be compared directly and discussed jointly.

Abstract

There are few methods for quantifying cell proliferation. Those tests describe the proliferation kinetics of a cell population, but they do not report the history of single cells, the number and frequency of cell divisions, or the precursor cell frequency. Cell-tracking assays based on dilution of the green fluorescent protein labelling dye, CFSE, has become the standard for monitoring cell proliferation. Other labelling dyes, e.g. CellTrace Violet and CellVue Claret, are also used for the same purpose. This study aimed to compare these three cell labelling methods for analysing the kinetics of cell viability, proliferation, and precursor cell frequency. Human peripheral blood mononuclear cells stimulated with Concanavalin A (ConA) were used as a model system. After labelling with a cell-tracking dye cells were divided into groups with and without ConA stimulation. From the 5th to 8th day, cells were collected and analysed with flow cytometry. Cell viability was not significantly different between labelled and unlabelled cells that received ConA stimulation. The proliferative fraction, proliferation index, and nonproliferative fraction were not significantly different among lymphocytes labelled with different dyes. Precursor cell frequency was also similar among cells labelled with the three cell-tracing dyes. The practical conclusion from our observations is that the results from cells labelled with different tracers may be compared directly and discussed jointly.

Get Citation

Keywords

cell proliferation; CFSE; CellTrace Violet; CellVue Claret; FACS analysis

About this article
Title

Monitoring cell proliferation in vitro with different cellular fluorescent dyes

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 51, No 3 (2013)

Article type

Original paper

Pages

193-200

Published online

2013-11-07

DOI

10.5603/FHC.2013.0027

Bibliographic record

Folia Histochem Cytobiol 2013;51(3):193-200.

Keywords

cell proliferation
CFSE
CellTrace Violet
CellVue Claret
FACS analysis

Authors

Joanna Zolnierowicz
Magdalena Ambrozek-Latecka
Jerzy Kawiak
Danuta Wasilewska
Grazyna Hoser

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