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Quality Control
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Data Management
Every frozen vial, testing sample, culture flask and reagent is individually bar coded for tracking purposes. Similarly every event from vial thawing, sample testing, cell line passage through to the freezing step is electronically recorded for each cell line, along with the batch of the consumables used, the specific equipment utilised and the identity of the handler. All quality control steps are recorded and must be passed before a cell line is to be released.
Mycoplasma Testing
All incoming cell lines and all batches of cells for storage are assessed for Mycoplasma infection. To test for Mycoplasma, medium samples are assessed using a commercial biochemical kit (MycoAlert, Cambrex) and an in-house PCR assay. The latter PCR is based on published primer sequence (Kong et al 2001, Appl Environ Microbiol 67(7): 3195-3200). Our detection methods have been validated for all Mycoplasma species that have been reported to infect cultured cell lines as well as most of the Mycoplasma species reported to infect humans. Mycoplasma can also be directly visualised in fixed cells using DAPI staining as an alternate form of detection.
Identification Testing
The identities of all human cell lines and all batches for storage 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 a commercial kit, which includes loci used by other cell banks such as the American Type Culture Collection for cell line authentication.
Sterility Testing
Every cell line is tested for the presence of contaminating micro-organisms upon culture in our facility and prior to freezing. Sterility testing of conditioned media in nutrient broths at specific temperatures ensures that our cell lines are free of fungal or bacterial contamination. Unless requested by researchers, all cell lines are cultured in the absence of antibiotic to ensure that antibiotic resistant or growth suppressed micro-organisms contaminations are detected.
Viability Testing
All cell line batches stored in liquid nitrogen are assessed for recovery and viability after cryopreservation, with viability determined 24 hours post-thaw. Sentinel cell lines are tested periodically for viability to confirm the integrity of liquid nitrogen storage long-term.
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