Human, mouse or rat? Species authentication of glioma-derived cell cultures

Cell culture and the use of cell lines are often fundamental requirements in basic scientific research. It is of the utmost importance for researchers to ensure that the cell lines in use have a well defined origin and are routinely re-analysed to highlight possible areas of contamination. In this preliminary study species specific primers were designed to easily distinguish between human, mouse and rat DNA with standard agarose gel electrophoresis. Inter-species contamination is often the most common form of contamination experienced, with the most common of cell lines in use being of human, mouse and rat derivation. A PCR-based assay was therefore developed to ensure an accurate, quick and cost effective means of determining any cell line contamination which could be easily executed on a routine basis. Furthermore, this simple PCR is able to identify the species in the inter-species mixture of DNA and therefore provides a valuable tool for the authentication of human cell lines.

Research highlights▶ Species specific primers have been designed to easily distinguish between human, mouse and rat DNA with standard agarose gel electrophoresis. ▶ The PCR assay was developed to ensure an accurate, quick and cost effective means of determining inter-species contamination of cell cultures on a routine basis. ▶ The PCR findings were cross-validated with species specific ICC using CD15 antibodies. ▶ Test-It™ Kit provides a valuable tool for the authentication of human cell lines.