Clonogenic assay of cells in vitro

Clonogenic assay or colony formation assay is an in vitro cell survival assay based on the ability of a single cell to grow into a colony. The colony is defined to consist of at least 50 cells. The assay essentially tests every cell in the population for its ability to undergo “unlimited” division....

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Bibliographic Details
Published in:Nature protocols Vol. 1; no. 5; pp. 2315 - 2319
Main Authors: Franken, Nicolaas A P, Rodermond, Hans M, Stap, Jan, Haveman, Jaap, van Bree, Chris
Format: Journal Article
Language:English
Published: London Nature Publishing Group UK 01.12.2006
Nature Publishing Group
Subjects:
Analytical Chemistry
Assaying
Biological assay
Biological Techniques
Biomedical and Life Sciences
Cell Culture Techniques
Cell death
Cell Proliferation - drug effects
Cell Proliferation - radiation effects
Cell survival
Cells
Cloning
Colonies
Colony-Forming Units Assay - methods
Computational Biology/Bioinformatics
Cytotoxic agents
Cytotoxicity
Humans
Ionizing radiation
Life Sciences
Methods
Microarrays
Organic Chemistry
Physiological aspects
protocol
Radiation
Radiation Dosage
Stains and staining (Microscopy)
Stem cells
Survival
Tumors
X-rays
Netherlands
ISSN:1754-2189, 1750-2799, 1750-2799
Online Access:Get full text
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Summary:Clonogenic assay or colony formation assay is an in vitro cell survival assay based on the ability of a single cell to grow into a colony. The colony is defined to consist of at least 50 cells. The assay essentially tests every cell in the population for its ability to undergo “unlimited” division. Clonogenic assay is the method of choice to determine cell reproductive death after treatment with ionizing radiation, but can also be used to determine the effectiveness of other cytotoxic agents. Only a fraction of seeded cells retains the capacity to produce colonies. Before or after treatment, cells are seeded out in appropriate dilutions to form colonies in 1–3 weeks. Colonies are fixed with glutaraldehyde (6.0% v/v), stained with crystal violet (0.5% w/v) and counted using a stereomicroscope. A method for the analysis of radiation dose–survival curves is included.
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ISSN:1754-2189
1750-2799
1750-2799
DOI:10.1038/nprot.2006.339