There are five major Cell Culture Authentication Databases:

- the American Type Culture Collection (ATCC)

- the German Collection of Microorganisms and Cell Cultures / DSMZ

- European Collection of Cell Cultures (ECACC)

- the Japanese Collection of Research Bioresources (JCRB)

- the RIKEN Bioresource Center Cell Bank / RIKEN

All use the standard for cell line authentication (validation and identification) which is STR Profiling.

Four of these five - DSMZ, ATCC, JCRB, and RIKEN have cooperated under the leading auspice of DSMZ to create a collaborative initiative, using a Consensus STR Panel, of an online Cell Verification Tool known as OSTRA (Online STR Analysis) for confirming the identity of any human cell line which allows easy multi-center interactive searches across all these databases.

The OSTRA tool can be accessed at:

http://www.dsmz.de/services/services-human-and-animal-cell-lines/online-str-analysis.html

This currently represents the most straightforward way to conduct authoritative authentication.

Constantine Kaniklidis

Director of Medical Research,

No Surrender Breast Cancer Foundation (NSBCF)

European Association for Cancer Research (EACR)

I don't have much experience here, but have you tried finding some labeled antibodies for protein markers on the cell surface? Some simple staining followed by flow cytometry should give a decent answer, I would think. Another option might be some quick PCR on the samples for a cell-specific gene.

Sorry I can't be more of a help.

For a flow and check for CD markers for the particular cell line. Easiest and quickest way!

Screening CD markers would be an option, if there is a database that provides information about correct CD markers for each cell line, right?

There are five major Cell Culture Authentication Databases:

- the American Type Culture Collection (ATCC)

- the German Collection of Microorganisms and Cell Cultures / DSMZ

- European Collection of Cell Cultures (ECACC)

- the Japanese Collection of Research Bioresources (JCRB)

- the RIKEN Bioresource Center Cell Bank / RIKEN

All use the standard for cell line authentication (validation and identification) which is STR Profiling.

Four of these five - DSMZ, ATCC, JCRB, and RIKEN have cooperated under the leading auspice of DSMZ to create a collaborative initiative, using a Consensus STR Panel, of an online Cell Verification Tool known as OSTRA (Online STR Analysis) for confirming the identity of any human cell line which allows easy multi-center interactive searches across all these databases.

The OSTRA tool can be accessed at:

http://www.dsmz.de/services/services-human-and-animal-cell-lines/online-str-analysis.html

This currently represents the most straightforward way to conduct authoritative authentication.

Constantine Kaniklidis

Director of Medical Research,

No Surrender Breast Cancer Foundation (NSBCF)

European Association for Cancer Research (EACR)

There are lists of CD markers for every cell line. Look in the literature.

What alternatives exist except CD marker analysis with FACS?

Just to get it straight: "How does YOUR LAB validate the identity of a cell line". Does your lab do this at all?

Hi Kai

We do STR profiling at our institute as Constantine suggested. It's done on every cell line we use, and is by far the current gold standard (short of sequencing the cell line). Most of the top journals have been making noises that they will (in the near future) require all cell lines to be authenticated by STR profiling prior to publication. As far as I'm aware there are two main STR databases and the kits you can buy will be compatible with one or the other.

Best of luck

Al

Alasdair is correct that STR Profiling is the now global standard and consensus for cell line characterization. The available technologies for establishing cell authentication and provenance - what I call jointly, cell "characterization" - include:

(1) Human leukocyte antigen (HLA) typing (as used in patient-donor tissue typing),

(2) karyotyping (visualization of chromosomal pattern and architecture),

(3) isoenzyme typing and

(4) perhaps the most sophisticated, DNA fingerprinting,

(5) STR Profiling

but all but STR Profiling are expensive and of limited reproducibility. In terms of practicality in the here and now, only the alternative technique of short tandem repeat (STR) profiling - the same PCR-based method I note being used by the FBI to populate the infamous CODIS (Combined DNA Index) database of forensic DNA samples) - is both manifestly feasible and affordable, being now a well-established and well-regarded technique for unambiguously characterizing a number of different loci in the human genome to provide an authoritative reference standard for human cell lines (as Eugene Elmore at the University of California (In Vitro Cell Dev Biol Anim. 2010) among others has convincingly argued, based on the demonstration by John Masters at University College London and colleagues (Proc Natl Acad Sci USA. 2001) that using STR profiling can effectively eliminate scientific misrepresentation due of cell line cross-contamination can be largely eliminated, and can serve to function as an international reference standard for human cell lines, a proposal I have advocated for some while.

At this time all five authoritative Cell Databases I identified - ATCC, DSMZ, ECACC, JCRB, and RIKEN - use STR Profiling, and four (ATCC, DSMZ, JCRB and Riken) have, as I noted above, pooled their resources to provide the first and online Cell Verification Tool, OSTRA (Online STR Analysis). However, ECACC also provides, besides STR Profiling, an option for cell line verification by DNA fingerprinting/barcoding, an option that is secondary but still available on request from the other Cell Database providers. Pricing of the service through OSTRA under DSMZ is standardized across the four internal database providers used (ATCC, DSMZ, JCRB, RIKEN) under DSMZ auspices (for "one-stop" shopping), but ECACC still maintains a distinct priceline although within the same small range as the others.

Alasdair is also right in noting the increasing pressure from journal publishers many of whom have initiated requests for proof of cell line characterization, although at the moment the AACR, ASCO/JCO, Nature and other high-impact groups will apparently accept either STR profiling (their preference) or DNA fingerprinting but not HLA typing, karyotyping, nor isoenzyme typing which have significant limitations I have documented elsewhere, and we should - long overdue - be seeing some standardization begin to appear across major journal groups on this issue which has caused many of them some significant embarrassment.

Constantine Kaniklidis

Director of Medical Research,

No Surrender Breast Cancer Foundation (NSBCF)

European Association for Cancer Research (EACR)

have you heard about multiplexion? they are in heidelberg and make specific tests for cell line authentication for example. www.multiplexion.de

Karyotype

Hi

We use STR analysis for 9 targets recommended by the ATCC database.

STR profiling is the most simple, straight forward and cost-effective way to determine if the cell line you are working with is what you think it is. All major repositories now have STR profile databases available for their cell lines for ease of comparison with what you have growing in your lab. In addition, if you are creating "in-house" cancer or stem cell lines, the best method to "authenticate" those lines would be to compare a DNA sample from the patient to the established cancer or stem cell line - this sets your "baseline" STR profile for all future testing with that cell line (for all future passages or if you give that line to colleagues for use). There are many service providers that offer very low cost testing with high quality - many of them work with international clients. I work for a service provider (Genetica in the USA) and we offer very competitive pricing with many different reporting options to suit your specific needs (whether just routine quality control or formal submission to journals for publication). Our website has a lot of information about the testing and a whole list of additional resources if you would like to read about the history of the problem of misidentification and cross-contamination. www.celllineauthentication.com

Hi Kai,

I hope you got some help by now but as Erin and others have pointed out, there are commercial services that make this fast, easy, and inexpensive with impeccable results. iQ Biosciences offers this service so if you need more technical info, you can see this webpage: http://www.iqbiosciences.com/bioservices/testing-services/cell-line-authentication or this youtube video with describes how the process is done: http://www.youtube.com/watch?v=-WCknRLzt0g

iQ Biosciences is very high throughput so unlike other services providers, can provide the highest service quality and lowest pricing for samples anywhere around the world. If you (or anyone reading this) would like to try a sample at no/minimal cost, please contact [email protected]. You will be impressed.

Best Regards,

Omar

Hi Kai,

Recently we noticed same thing in my lab too. I recommend to go for STR analysis. And don't go to higher passage number also.  If  STR is fine only then continue with the  experiments.

 Please follow these links.

 https://www.atcc.org/STR_ Database.aspx

https://www.atcc.org/Products/ Cells_and_Microorganisms/Cell_ Lines/Misidentified_Cell_ Lines.aspx

 http://iclac.org/databases/ cross-contaminations/

This is the address of Cell Line Authentication Services :

(Cell Line Authentication Services

Genetica DNA Laboratories - a LabCorp brand

[email protected]

1-800-IDENTITY (433-6848)

Direct Line:  1-513-206-1653

www.celllineauthentication.com

"Authenticate Today to Publish Tomorrow")

I hope this will help.

Thanks

Kiran