Increased efficiency for performing colony formation assays in 96-well plates: novel applications to combination therapies and high-throughput screening

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Original Research Paper

Increased efficiency for performing colony formation assays in 96-well plates: novel applications to combination therapies and high-throughput screening

David Katz, Emma Ito, Ken S. Lau, Joseph D. Mocanu, Carlo Bastianutto, Aaron D. Schimmer, and Fei-Fei Liu

BioTechniques 44:Pix-Pxiv (February 2008) doi 10.2144/000112757

Review

The colony formation assay (CFA) is the standard assay for measuring the effects of cytotoxic agents on cancer cells in vitro. Traditionally, this assay is performed in 6-well plates, which is time consuming, particularly when evaluating multiple compounds, in combination, in a single assay.

This paper, from a group in Canada, describes an automated method for improving the throughput of the CFA by incorporating high content analysis – or automated microscopy – and software analysis so that 96-well plates can be used. Two types of human cancer cell lines were analyzed after treatment with ionizing radiation in combination with anti-cancer compounds. Colonies were stained with a two color technique. By using this high throughput assay, several compounds from a known small molecule library were identified as “hits” in a rapid, automated program and these compounds were confirmed as active in separate assays. This CFA format could lend itself to evaluation of multiple permutations of combination therapies, such as radiation and drug treatments, thus speeding up the overall evaluation process for such treatments in cancer research.

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  Figure 1. Reproducibility of a 96-well colony formation assay (CFA) compared with a traditional 6-well CFA. (A) CFAs were performed on FaDu cells using the 96-well and the 6-well assays. (B) CFAs were performed on A549 cells using the 96-well and the 6-well assays. Each datum represents the mean ± sem from three independent experiments for 6-well data and two independent experiments for 96-well data.