Cross-Contaminated & Misidentified Cell Lines

Instances of cross-contamination between cell lines are more common than generally appreciated. Surveys of the scientific literature have suggested that up to 20% of published work based on the use of human cell lines specifies lines that are known to be mis-identified^1,2.

This page aims to draw attention to those cell lines that are known, or suspected to be misidentified. To view an extensive list of misidentified cell lines compiled by Dr. R. Ian Freshney and Dr. Amanda Capes-Davis click here (opens in a new window). This list has been compiled from published reports in the scientific literature and details obtained from cell banks, their websites, and Wikipedia. Please also view the Nardone Open Letter which should be of concern to all individuals and organisations using cell cultures.

The information below is specific to cell lines affected by misidentification or cross-contamination which are either currently available in, or were previously available and now withdrawn from, the ECACC catalogue. Where cell lines continue to be available their description in the ECACC catalogue highlights their determined identity which supersedes the original assumed origin.

1. HeLa Derivatives

The cell lines listed in Table 1 below have been found to be genetically indistinguishable from the human cervix carcinoma cell line HeLa by short tandem repeat (STR)-PCR profiling. Therefore, these cell lines must be considered to be HeLa. To highlight this, the original cell line name has had ‘(HeLa derivative)’ appended to it where these cell line names appear in the ECACC catalogue.

Such cell lines should be considered as derived from cervical tissue and should not be used in studies to represent any other type of tissue, including that wrongly assigned to the misidentified cell line.

Table 1. Cell lines that are known to be derived from the cervix carcinoma cell line HeLa. The cross-contamination occurred prior to deposit with ECACC and affects all known sources of these cell lines.

2. Cell lines with Re-assigned Identities (Non-HeLa Derivatives)

Table 2. Cell lines that have been found to be cross-contaminated with or mis-identified as another cell line and as a result have had their identities reassigned. Please note that these re-assignments affect all official sources of the cell lines available. The cross-contamination occurred prior to deposit with ECACC.

3. Identification Queries

U-373 MG

The authenticity of some sources of the U-373 MG cell line has been questioned⁶. The ATCC reported that their stock of U-373 MG has been shown to have differing genetic properties to stock from the originators laboratory, and to share similarities with another glioblastoma cell line, U-251. In light of this, ECACC suspended supply of the U-373 MG cell line and undertook an investigation into the authenticity of its own stock. ECACC found similar results to the ATCC i.e. the stock held as U-373 MG was found to be identical by short tandem repeat (STR)-PCR profiling to U-251. The U-373 MG cell line listed under catalogue number 89081403 has been re-named as ‘U-251 (formally known as U-373 MG)’ and allocated a new ECACC catalogue number: 09063001. A new deposit of U-373 MG has been obtained from the originator and shown to differ from U-251 by STR-PCR profiling, this has been added to the collection and is called U-373 MG (Uppsala), ECACC catalogue number 08061901, to distinguish it from the U-373 MG cell line which was listed under catalogue number 89081403 – which is now known to be U-251. Both cell lines are derived from glioblastoma astrocytomas.

IPTP/98

ECACC has recently determined, using isoenzyme species analysis, that the IPTP/98 cell line sample deposited with ECACC is of rat origin rather than human. Although the sample received was purported by the depositor to be a human glioblastoma cell line an investigation has shown that the original material received was in fact of rat origin. As a result of this discovery IPTP/98 has been withdrawn from the ECACC catalogue.

IPDDC-A2

ECACC has recently determined, using isoenzyme species analysis, that the IPDDC-A2 cell line sample deposited with ECACC is of rat origin rather than human. Although the sample received was purported by the depositor to be a human astrocytoma cell line an investigation has shown that the original material received was in fact of rat origin. As a result of this discovery IPDDC-A2 has been withdrawn from the ECACC catalogue.

TEC61

ECACC has recently determined using STR-PCR profiling that TEC61, ECACC cat no. 06031505, which was claimed to be derived from thyroid endothelium is actually JEG3, a human choriocarcinoma. The cell-conditioned medium from JEG3 was used to support the growth of the putative endothelial cell line (Patel et al., 2009 PMID: 12388152, retraction - PMID:19625678)⁷. As a result of this discovery, TEC61 has been withdrawn from the ECACC catalogue.

CHP-100

ECACC has recently determined using STR-PCR profiling that CHP-100, ECACC cat no. 06122001, which was claimed to be a neuroblastoma is actually IMR 32 another neuroblastoma⁸. As a result of this discovery, CHP-100 has been withdrawn from the ECACC catalogue.

If additional misidentified cell lines are discovered ECACC will report findings on this page.

References

1. MacLeod et al., (1999) Widespread intraspecies cross-contamination of human tumor cell lines arising at source. Int J Cancer 83(4) 555-563 (Cell line: ECV304)
   
2. Drexler HG et al. (2003) False leukemia-lymphoma cell lines: an update on over 500 cell lines. Leukemia, 17(2):416-426.
   
3. Dirks et al., (1999) ECV304 (endothelial) is really T24 (bladder carcinoma): cell line cross- contamination at source In Vitro Cell Dev Biol Anim 35: 558-559 (Cell line: ECV304)
   
4. Stratton et al., (1989) Misidentified Cell. Nature;337:311; (Cell line: TE671)
   
5. Chen et al., (1989) Cell Identity Resolved. Nature;340:106; (Cell line: TE671)
   
6. ATCC website & Ishii et al., (1999) Frequent Co-Alterations of TP53, p16/CDKN2A, p14ARF, PTEN Tumor Suppressor Genes in Human Glioma Cell Lines. Brain Pathol 9: 469-79, (Cell line: U-373 MG)
   
7. Patel et al., 2003 Isolation and characterization of human thyroid endothelial cells. Am J Physiol Endocrinol Metab. 2003 Jan;284(1):E168-76. PMID: 12388152 Retraction (2009): PMID:19625678 (Cell line: TEC61)
   
8. Reported by the European Collection of Cell Cultures (ECACC) on the Health Protection Agency Culture Collections website. (Cell line: CHP-100)