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-identified1,2.
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misidentification or cross-contamination which are either currently available in, or were previously available and now withdrawn from, the ECACC catalogue.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.
| Cell Line | ECACC cat no. | Determined to be: | Claimed tissue of origin: |
| C16 (HeLa derivative) | 84121902 | HeLa derivative, human cervix carcinoma cell line | Human foetal lung |
| Chang Liver (HeLa derivative) | 88021102 | HeLa derivative, human cervix carcinoma cell line | Human liver |
| Clone 1-5c-4 (HeLa deriative) | 88021103 | HeLa derivative, human cervix carcinoma cell line | Human conjunctiva |
| D98/AH2 Clone B (HeLa derivative) | 85112701 | HeLa derivative, human cervix carcinoma cell line | Somatic cell hybrid (HeLa cells and mouse 3T3 cells) |
| FL (HeLa derivative) | 90111910 | HeLa derivative, human cervix carcinoma cell line | Human amnion |
| GIRARDI HEART (HeLa derivative) | 93120822 | HeLa derivative, human cervix carcinoma cell line | Human heart |
| Hep-2C (HeLa derivative) | 85020207 | HeLa derivative, human cervix carcinoma cell line | Human larynx |
| Hep2 (Clone 2B) (HeLa derivative) | 85011412 | HeLa derivative, human cervix carcinoma cell line | Human larynx |
| Hep2 (HeLa derivative) | 86030501 | HeLa derivative, human cervix carcinoma cell line | Human larynx |
| HSG (HeLa derivative) | 95031024 | HeLa derivative, human cervix carcinoma cell line | Human sub-mandibular gland |
| INT 407(HeLa derivative) | 85051004 | HeLa derivative, human cervix carcinoma cell line | Human embryonic intestine |
| JIII (HeLa derivative) | 93120824 | HeLa derivative, human cervix carcinoma cell line | Human monocytic leukaemia |
| KB (HeLa derivative) | 94050408 | HeLa derivative, human cervix carcinoma cell line | Human oral epidermis |
| L-41(HeLa derivative) | 96121716 | HeLa derivative, human cervix carcinoma cell line | Human bone marrow |
| L132 (HeLa derivative) | 89111004 | HeLa derivative, human cervix carcinoma cell line | Human embryonic lung |
| WISH (HeLa derivative) | 88102403 | HeLa derivative, human cervix carcinoma cell line | Human amnion |
| WKD (HeLa derivative) | 93120839 | HeLa derivative, human cervix carcinoma cell line | Human conjunctiva |
| WRL 68 (HeLa derivative) | 89121403 | HeLa derivative, human cervix carcinoma cell line | Human embryonic liver |
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)
| Cell Line | ECACC cat no. | Determined to be: | Originally referred to as: |
| ECV304 | 92091712 | identical to T24 - a human bladder carcinoma epithelial cell line1,3 | ‘spontaneously transformed’ human umbilical cord endothelial cells |
| TE671 | 89071904 | a derivative of the rhabdomyosarcoma cell line RD4,5 | a medulloblastoma cell line |
| TE671 Subline No.2 | 94052610 | a derivative of the rhabdomyosarcoma cell line RD4,5 | a medulloblastoma cell line |
Table 2. Cell lines that have been found to be cross-contaminated with 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 have 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 is undertaking an investigation into the authenticity of its own stock of the U-373 MG. U-373 MG will remain unavailable from ECACC until this investigation is complete.
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 IPTP98 has been withdrawn from the ECACC catalogue.
If additional misidentified cell lines are discovered ECACC will report findings on this page.
References
- 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)
- Drexler HG et al. (2003) False leukemia-lymphoma cell lines: an update on over 500 cell lines. Leukemia, 17(2):416-426.
- 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)
- Stratton et al., (1989) Misidentified Cell. Nature;337:311; (Cell line: TE671)
- Chen et al., (1989) Cell Identity Resolved. Nature;340:106; (Cell line: TE671)
- 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)
