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A simple, informative, and quantitative flow cytometric method for assessing apoptosis in cultured cells
Robert C Warrington1, Joanne N Norum, Jeffrey L Hilchey
1Department of Biochemistry, College of Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, Canada. warrington@sask.usask.ca
Progress in Neuro-Psychopharmacology & Biological Psychiatry
|March 27, 2003
Summary
This study introduces a new flow cytometry method to easily quantify apoptosis in cultured cells. The technique uses DNA fluorescence and side-scatter signals to reveal more information than traditional methods.
Area of Science:
- Cell Biology
- Biotechnology
- Cancer Research
Background:
- Apoptosis assessment is crucial for understanding cell death and drug efficacy.
- Conventional flow cytometry methods for apoptosis analysis can be limited in the information they provide.
- There is a need for facile, informative, and quantifiable methods to study apoptosis in diverse cell types.
Purpose of the Study:
- To describe a novel, highly informative, and quantifiable method for assessing apoptosis in tissue culture cells.
- To demonstrate the utility of this method in evaluating the effects of antineoplastic agents and other compounds on apoptosis.
- To showcase the advantages of correlated two-parameter flow cytometric analysis for apoptosis studies.
Main Methods:
- Utilized fixed, propidium iodide-stained cells for analysis.
- Employed correlated two-parameter flow cytometry, analyzing log(10)DNA fluorescence signals versus log(10) side-scatter (SS) signals.
- Applied the method to various cultured cell types, including epithelial, fibroblast, and myeloid cells.
Main Results:
- The novel method provided substantially more information than conventional analyses.
- Demonstrated the effects of cisplatinum (cisP) and camptothecin (CAM) on apoptosis in multiple cell lines.
- Showed L-histidinol to be pro-apoptotic in CCRF-CEM cells and to modulate CAM-induced apoptosis in B16f10 melanoma and 3T3 fibroblasts.
Conclusions:
- The described method is facile, highly informative, readily quantifiable, and generally applicable for apoptosis assessment.
- Correlated two-parameter flow cytometry offers significant advantages over conventional methods for studying apoptosis.
- This technique is valuable for monitoring drug effects and understanding cellular responses to apoptotic stimuli.