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Related Experiment Video

Updated: Jul 2, 2026

Enumeration of Neural Stem Cells Using Clonal Assays

Enumeration of Neural Stem Cells Using Clonal Assays

Published on: October 4, 2016

Don't look: growing clonal versus nonclonal neural stem cell colonies.

Brenda L K Coles-Takabe¹, Ian Brain, Kelly A Purpura

¹Departments of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.

Stem Cells (Dayton, Ohio)

|September 2, 2008

Summary

The neurosphere assay

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Last Updated: Jul 2, 2026

Enumeration of Neural Stem Cells Using Clonal Assays

Enumeration of Neural Stem Cells Using Clonal Assays

Published on: October 4, 2016

Neural-Colony Forming Cell Assay: An Assay To Discriminate Bona Fide Neural Stem Cells from Neural Progenitor Cells

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Published on: March 6, 2011

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Published on: December 16, 2017

Area of Science:

Neuroscience
Stem Cell Biology

Background:

The neurosphere assay is widely used for neural stem cell (NSC) research.
Recent studies question the clonality of this assay for quantitative NSC assessment.

Purpose of the Study:

To rigorously evaluate the clonality of the neurosphere assay.
To identify factors influencing clonal sphere formation.

Main Methods:

Culturing mixtures of differentially labeled neural cells.
Video microscopy to track single neural cell proliferation.
Encapsulation of single neural stem cells (NSCs) and their progeny.

Main Results:

Clonal colonies formed from primary cells at ≤10 cells/µL.
Passaged NSCs required ≤1 cell/µL for clonal spheres.
Plate movement significantly increased non-clonal colonies.

Conclusions:

Clonality in neurosphere assays is sensitive to cell density and handling.
Avoiding movement and using single-cell encapsulation ensures clonal neurosphere formation.
This work refines methods for reliable neural stem cell quantification.