Cross-contamination and misidentified cell lines affect approximately one-fifth of biomedical research. Consequently, scientists may believe or claim that they are working with cells derived from one individual or animal species, only to later learn that the cells were derived from a different individual or species altogether.
To provide researchers with confidence in the validity and integrity of their work, and to meet the requirements for acceptance of research publications in peer-reviewed journals, we offer cell line identification services to authenticate human cell lines. As part of Garvan’s NATA ISO 17025 accredited Molecular Genetics Core Facility, our high-throughput, accurate and cost-effective services support researchers within the Research Precinct, as well as external clients.
We offer three key cell line identification services:
Cell Line Identification and Authentication (human)
Cell line Identification and Authentication (mouse).
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Cell line identification and authentication (human)
Short Tandem Repeat (STR) loci consist of short, repetitive sequence elements that are 3–7 base pairs in length. The number of repeats in STR loci can be highly variable among individuals, making these genetic markers effective for human identification or cell line authentication purposes.
Our cell line identification services uses the PowerPlexR 18D System, which has 18 markers (seventeen STR loci and Amelogenin). After analysis, cell-line IDs are considered to be a match if their profiles are more than 80% identical. The choice of >80% as a suitable threshold is based on published work from the ASN-0002 Standard workgroup, looking at the eight core loci in related cell line samples from five cell banks.
This service includes an authentication certificate that will identify reference sample IDs via Garvan’s cell line identification database.
Mycoplasmas are eight species in the Mollicutes class that collectively account for over 95% of isolates from cultured cells (Barile, 1979; McGarrity and Kotani, 1985; Btilske, 1988; McGarrity et al., 1992). Our service detects these mycoplasmas, which are: Mycoplasma hyorhinis, Mycoplasma orale, Mycoplasma hominis, Mycoplasma arginini, Mycoplasma salivarium, Mycoplasma pirum, Mycoplasma fermentans and Acholeplasma laidlawii.
In addition to this, we are able to detect: Mycoplasma pneumoniae, Mycoplasma gallisepticum, Mycoplasma genitalium, Mycoplasma penetrans, Mycoplasma synoviae, Mycoplasma bovis, Mycoplasma hyopneumoniae, Ureaplasma urealyticum and Spiroplasma citri.
We perform four polymerase chain reactions (PCRs) with primers that amplify a portion of the 23S rRNA gene and a conserved region within the 16S rRNA gene. These primers do not detect eukaryotic DNA or bacterial genera with close phylogenetic relation to mycoplasmas, such as Clostridium, Lactobacillus and Streptococcus. Please submit 1 ml of media for this test (with or without cells).
Cell line authentication (mouse)
Our Mouse Cell Line Authentication Service uses a panel that was developed by the Consortium for Mouse Cell Line Authentication, to validate STR markers for intraspecies identification of mouse cell lines. The assay we use targets 19 mouse STRs (Interlaboratory study to validate a STR profiling method for intraspecies identification of mouse cell lines, PLoS ONE 14(6): e0218412).
While there are validated methods, kits and databases available for identity testing for human cells, there are no commercial kits available for mouse cell lines. The assay we use improves upon patented methods (US Patent 9,556,482 B2), which include additional primers that target a total of 19 mouse STR loci and two human STR markers for contamination detection (US patent application publication 2017/0101677 A1).
Our services are provided through our sample submission portal. Visit the portal to create a new account, log into an existing account or find out more about submitting samples and accessing results. You can submit DNA or cells for the Cell Line Identification Service. For mycoplasma testing, we recommend submitting supernatant.
Before you submit a sample, please read the cell line identification service sample submission guidelines for information about labelling your samples and submission requirements. The turnaround time is a maximum of 10 working days, however, samples are usually processed within a week.
Please mail samples to:
Garvan Molecular Genetics
Garvan Institute of Medical Research
384 Victoria Street
Darlinghurst NSW 2010
For support in accessing the portal, please email firstname.lastname@example.org.
Information about pricing is available here. Garvan researchers are eligible for a discounted price. Please visit Facilities on the intranet for more information.
Garvan's Genetics Core Facility was established in 2004 through support from the Australian Cancer Research Foundation (ACRF).