Related Experiment Video
Updated: May 12, 2026
06:26
Analysis of Side Population in Solid Tumor Cell Lines
Published on: February 23, 2021
Flow cytometry of the side population (SP).
1Vall d'Hebron Institut de Recerca, Hospital Universitari Vall d'Hebron, Universitat Autònoma de Barcelona, Barcelona, Spain.
Current Protocols in Cytometry
|April 3, 2013
Summary
Side population (SP) stem cells are crucial for research and isolation. New methods using Vybrant DyeCycle Violet (DCV) stain offer improved discrimination over older Hoechst 33342 (Ho342) assays for stem cell detection.
Area of Science:
- Cell Biology
- Stem Cell Research
- Biotechnology
Background:
- Side population (SP) cells are a key stem cell marker.
- SP cells are identified by their ability to efflux fluorescent dyes like Hoechst 33342 (Ho342).
- Traditional Ho342 assays present challenges in experimental setup and data analysis.
Purpose of the Study:
- To detail methods for identifying and isolating stem cells using the side population.
- To introduce and explain the Vybrant DyeCycle Violet (DCV) assay as an alternative to Ho342.
- To provide guidance for optimizing SP cell detection and analysis.
Main Methods:
- Utilizing fluorescence-activated cell sorting (FACS) for cell isolation.
- Employing Hoechst 33342 (Ho342) dye efflux for SP cell identification.
- Implementing the Vybrant DyeCycle Violet (DCV) stain assay for SP cell discrimination.
Main Results:
- Side population (SP) cells are characterized by CD34(-) status and dye efflux properties.
- The Ho342 assay, while effective, requires specialized expertise and careful condition setting.
- The DCV assay provides a documented alternative for discriminating SP cells.
Conclusions:
- SP analysis is vital for stem cell research and isolation.
- The DCV assay offers a potentially more accessible method for SP cell detection.
- This work provides practical guidance for researchers using SP assays.
