 The facility performs three forms of quality control. Firstly, all incoming cell lines and all batches of cells for storage are assessed for Mycoplasma and other contaminating micro-organisms. To test for Mycoplasma, medium samples are routinely assessed using two test methods. One of these is commercially available (MycoAlert® Mycoplasma Detection Kit, Lonza), while the second method is an in-house PCR assay. The PCR primers used detect the six Mycoplasma species commonly encountered in cell culture (Kong et al 2001, Appl Environ Microbiol 67(7): 3195-3200) and can detect femtogram quantities of Mycoplasma DNA. Where a clear result cannot be determined, Mycoplasma can be enriched by culture on an indicator cell line.
The identities of all human cell lines are confirmed using short tandem repeat (STR) profiling. This technique is widely used for forensic identification and has been recommended as best practice for authentication of cell lines worldwide (Masters et al 2001, Proc Natl Acad Sci USA 98(14): 8012-7). CellBank Australia uses Applied Biosystems' AmpFLSTR® Identifiler™ amplification kit, which detects the CODIS loci used by forensics laboratories. Loci are also used by other repositories such as the American Type Culture Collection (ATCC), which means that cell lines can be directly compared to those in other collections.
Finally, all batches stored in liquid nitrogen are assessed for recovery and viability after cryopreservation. Sentinel cell lines are also tested periodically to confirm the integrity of liquid nitrogen storage long-term. |