acumen: high throughput quantification of hepG2 cell colony formation in soft agar

Research to identify new anticancer drugs is currently facing significant challenges, as only 5% of compounds that show efficacy in preclinical development go on to become licensed drugs. Traditionally 2D cell culture models have been employed to evaluate drug candidates in the early phases of the drug discovery process, however, there is increasing evidence that cells grown in 2D monolayers do not accurately reflect the biological complexity of tumours. The requirement for better in vitro tumour models has led to the development of 3D cell culture models, which retain many of the morphological and genetic traits of tumours.

Enumeration of colonies has been traditionally carried out using a semi-solid agarose bilayer system in Petri dishes and involves manual counting on a microscope. High content analysis can be used to determine additional parameters such as colony size, shape, or fluorescence intensity, however, this approach has proved to be complicated due to the technical limitations of many CCD-based imaging systems…