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Characterization of neurosphere cell phenotypes by flow cytometry
1Cancer Center and Cell Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA. Ruud.Hulspas@umassmed.edu
Cytometry
|July 6, 2000
Summary
Neurosphere cultures contain a subpopulation of neural progenitor cells characterized by low Hoechst 33342 fluorescence. Purifying this subpopulation yields cells resembling freshly isolated neural cells.
Area of Science:
- Neuroscience
- Stem Cell Biology
Background:
- Neurosphere cultures are vital for neural stem cell research.
- Current assays indicate a low number of neural progenitors in these cultures.
Purpose of the Study:
- Characterize neurosphere cultures.
- Define subpopulations for neural progenitor cell purification.
Main Methods:
- Embryonic mouse neurosphere cells stained with Hoechst 33342.
- Flow cytometry used for analysis and sorting based on fluorescence.
- Sorted subpopulations reanalyzed after 7 days.
Main Results:
- Neurosphere cultures contain a high number of weakly Hoechst-stained cells.
- This subpopulation, when cultured separately, generates cells similar to fresh embryonic brain cells.
- The purified subpopulation shows fewer weakly Hoechst-stained cells upon re-culture.
Conclusions:
- Neurosphere cultures harbor a distinct subpopulation with low Hoechst fluorescence.
- This subpopulation can be purified and generates cells with a phenotype similar to uncultured neural cells.