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Clonogenic Assay: Adherent Cells

Clonogenic Assay: Adherent Cells

Published on: March 13, 2011

Clonogenic assay: adherent cells.

Haloom Rafehi¹, Christian Orlowski, George T Georgiadis

¹Epigenomic Medicine, BakerIDI Heart and Diabetes Institute, The Alfred Medical Research and Education Precinct.

Journal of Visualized Experiments : Jove

|March 30, 2011

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The clonogenic assay measures cell reproductive viability after treatments like radiation. This study details the assay for adherent cells and shows how an antioxidant can modify radiation response.

Area of Science:

Cell Biology
Radiation Biology
Cancer Therapeutics

Background:

The clonogenic assay, established over 50 years ago, assesses cell reproductive viability.
It's a cornerstone in radiation biology for evaluating cell sensitivity and treatment efficacy.
The assay quantifies a single cell's capacity to form a colony of 50+ cells.

Purpose of the Study:

To demonstrate the clonogenic assay procedure for adherent cell lines.
To detail the calculation of plating efficiency and survival fractions post-radiation.
To exemplify radiation-response modification using a natural antioxidant.

Main Methods:

Treatment of cell monolayers in tissue culture flasks.
Preparation of single-cell suspensions and plating in petri dishes.

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Incubation, fixation, and staining of colonies to assess survival.

Main Results:

The study provides a general protocol for performing the clonogenic assay with an immortalized human keratinocyte cell line (FEP-1811).
It outlines the calculation of key metrics like plating efficiency and survival fraction.
Demonstrated modification of radiation response by a natural antioxidant formulation.

Conclusions:

The clonogenic assay is a robust method for assessing cell survival and treatment effects.
The protocol is applicable to adherent cell lines, including human keratinocytes.
Natural antioxidants show potential in modulating cellular responses to radiation therapy.