ECACC offers an authentication service to verify the identity and provenance of cell lines; the consequences of working with a cell line that is misidentified or cross contaminated can be profound.
If cell cultures are not what they are reported to be then work can be invalidated and resources wasted.
Instances of cross contamination are more common than generally appreciated.
Estimates suggest that up to one in five experiments in fields such as cancer and microbiology employ the wrong cells.
ECACC recommends authenticating cell lines prior to commencing work and re-authenticating cell lines at regular intervals.
If you are generating new cell lines it is advisable to define their DNA profiles at an early stage.
ECACC can advise on an authentication programme to support individual customer needs.
Authenticity of a cell line can be assured within four weeks at a relatively low cost.
Methods of authentication available at ECACC.
Isoenzyme analysis.
Isoenzyme analysis looks at the enzymes malate dehydrogenase, glucose-6 phosphate dehydrogenase, nucleoside phosporylase and lactate dehydrogenase which are present in all species but show hetrogenicity which can be seen by electrophoresis.
By analysing these four isoenzymes and comparing the migration patterns with known standards and controls it is possible to speciate cell lines.
DNA fingerprinting.
A powerful molecular tool for the validation of cell lines.
Multilocus DNA fingerprinting produces a complex banding pattern unique to that cell line.
This technique enables the demonstration of continuity between master and working banks and identifies cross contamination and instability of cell lines using the multilocus DNA fingerprint library established at ECACC.
This technique can be used for most animal species.
Multiplex PCR.
A quick and powerful tool for the validation of human cell lines only.
This semi-automated system for DNA profiling uses up to ten primer sets to produce a unique banding pattern that can be compared with archived profiles on a database.
Like multilocus DNA fingerprinting, the technique can be used to demonstrate continuity between master and working banks and detect contamination and instability in cell lines.
DNA profiling in itself only confirms the continuity of a cell line, it does not speciate cells.
Such characterisation is carried out by isoenzyme analysis and is usually carried out in conjunction with DNA profiling to fully authenticate cells.
