Genotyping and Authentication of Cell Lines
Published studies with incorrectly assigned cell lines are one of the causes of non-reproducible scientific data. For this reason, among others, the NIH has made recommendations that should help to improve this situation (www.nih.gov/research-training/rigor-reproducibility/principles-guidelines-reporting-preclinical-research). This includes the authentication of cell lines, i. e. confirmation of the use of the correct cell line by genotyping, as well as proof of the absence of contamination, e. g. with mycoplasmas or other (e. g. human) cell lines.
Basically, one method has become established, genotyping by means of STR markers (microsatellites). For human cell lines, the identical STR kits are used as in forensics. These have proven to be very effective. Consequently, there is an effort to use this type of genotyping for other cell lines as well. However, other methods are also available, such as karyotyping, whole genome sequencing, specific probes, CO1 barcoding or SNP genotyping.
We have STR sets for genotyping of mouse, rat as well as human cell lines.
The STR genotyping set of the GVG is also able to distinguish all offspring of one parent pair of an inbred strain from each other, i. e. also between individuals of C57BL/6J. This is exactly what is used for authentication to obtain individual STR profiles. From the marker set, the most appropriate STR marker was identified per chromosome and these were combined into an authentication multiplex for cell lines. Thus, there is a high probability that your newly generated cell line will have a unique, specific genetic fingerprint.
As a quality assurance measure for mouse cell lines, markers specific to defined inbred strains of mouse are genotyped. These include indels (insertion-deletion polymorphisms) as well as STR markers, which have strain-specific alleles. The sex is also determined. This is used to check whether the designated inbred strain of the mouse is actually present. In addition, the cell line is prooved for possible contamination, e. g. with mycoplasma or cell lines foreign to the species (e. g. human cell lines).
The STR set for authentication of mouse cell lines also offers the possibility for genetic monitoring of changes in the cell line depending on the number of passages performed. In this way, new mutations can be detected, which indicate that gene drift has occurred (see also question: Why more than 2 alleles per marker).
If the same cell line is further grown in different laboratories, such changes will not occur in the same STR markers and the genetic profiles of the cell line between the two laboratories will differ slightly over time. This would be an indication of gene drift that has already occurred.
The processing time is 10 working days after receipt of the sample. As a rule, however, the result is communicated much earlier.
The report includes the identity of the inbred strain, the sex of the animal from which the cell line was generated, and the specific genetic fingerprint based on the results of STR genotyping.
In principle, a frozen pellet of the cell line can be submitted. Ready extracted DNA from commercial extraction kits (no "quick-and-dirty" extractions) works equally well. In this case, please send us half of the extract and keep the other half as a reserve sample.
Please always ship samples refrigerated, either on ice or ideally on dry ice, preferably early in the week so that samples are not kept over the weekend without cooling.
Our sample supply note should be used for reliable assignment and for the unambiguous naming of samples (in the report). Please fill out the template accordingly and send it to us electronically.
We do not provide any packaging materials. We recommend using simple 1.5 ml Eppendorf tubes for the samples.
Delivery to GVG can be made to the attention of Mrs. Schmidt (Tel. 0341- 392 859 21, GVG Genetic Monitoring GmbH, Deutscher Platz 5e, 04103 Leipzig). For shipping you can use our DHL account (DHL Express #144005514). In this case, we will invoice you for the transport costs together with our analysis service. Just call your local representative from DHL Express to have your samples picked up.
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