Misidentified Cell Lines

The implementation of STR analysis as part of routine authentication procedures has led to the discovery that some human cell lines deposited to ATCC are misidentified. STR analysis allows for verification of human cell line identity, and is now routinely performed by ATCC during accessioning and replenishment of cultures to avoid distribution of misidentified cell lines to the scientific community.

Authentication Technology Applied at
ATCC Reveals Misidentified Cell Lines

As a result of implementing STR analysis (DNA profiling) as part of ATCC routine authentication procedures, ATCC discovered that some human cell lines are not as originally reported by the depositor (see below). These cell lines were deposited many years before such procedure was available. See the complete list.

ATCC STR analysis uses multiplex PCR to simultaneously amplify eight STR loci plus amelogenin for gender determination. These loci are among the most informative polymorphic markers in the human genome (D16S539, D7S820, D13S317, D5S818, CSF1PO, TPOX, TH01, vWA). A unique DNA pattern of repeating units is generated for each human cell line analyzed. STR analysis is critical today to verify the identity of human cell lines and is now routinely performed at the time of accessioning a new cell line and at the replenishment of each distribution stock to avoid misidentified cell lines before they are distributed to the scientific community. The DNA profile of each replenished stock is verified by comparing to the baseline profile of the token stock derived from the depositor.

Inappropriate Y – DNA profiling at ATCC for amelogenin, a sex-chromosome-specific PCR assay that can distinguish X chromosome-specific products from Y chromosome-specific products revealed the presence of Y chromosomes in six human cell lines of putative female origin. Confirmation of the general findings was accomplished by QM staining, C-banding, and FISH, with a whole chromosome paint probe to the human Y chromosome.

The cell lines include:

 Designation

ATCC® No.

 Cytogenetic
Confirmation

 OV-1063  CRL-2183™  QM, C, FISH
 CHP-234  CRL-2272™  
 NCI-H738  CRL-5839™  QM, C, FISH
 NCI-H1514  CRL-5873™  QM, C, FISH
 NCI-H1622  CRL-5880™  
 HBL-100  HTB-124™  QM, C, FISH

ATCC discontinued distribution of these lines following an investigation which uncovered no compelling reasons not to do so.

HeLa Derivatives: — ATCC strongly advises against the use of these cell lines as models for original source material. Published data from researchers and testing performed during ATCC accessioning have established that these cell lines are contaminated with HeLa cells. These findings are based on isoenzyme analysis, HeLa marker chromosomes and DNA fingerprinting. HeLa contaminated cell lines should not be chosen for study when the specific organ or tissue of presumptive origin is of importance to the validity of the research as results can be compromised.

These cell lines include:
Designation ATCC® No.
L132 CCL-5™
Intestine 407 CCL-6™
Chang Liver CCL-13™
KB CCL-17™
clone 1-5c-4 CCL-20.2™
AV-3 CCL-21™
HEp-2 CCL-23™
WISH CCL-25™
FL CCL-62™

Identities in Question:

MDA-MB-435S (ATCC® HTB-129™) — ATCC advises against the use of this cell line as a model for original source material. Recent studies have generated questions about the origin of the parental cell line, MDA-MB-435, and by extension MDA-MB-435S and its derivatives listed below. Gene expression analysis of the cells produced microarrays in which MDA-MB-435 clustered with cell lines of melanoma origin instead of breast (PubMed IDs: 10700174, 1510101, 15679052).

Additional studies have since corroborated a melanocyte origin of MDA-MB-435, to which ATCC has responded by pursuing its own investigation into the identity of this cell line. The cell line to which MDA-MB-435 is reported to have been cross-contaminated with is the M14 melanoma line (PubMed IDs: 12354931, 17004106).

Derivatives of MDA-MB-435S with identifies in question are:

Designation ATCC® No.
M4A4 CRL-2914™
M4A4 GFP CRL-2915™
M4A4 LM3-2 GFP CRL-2916™
M4Af LM3-4 CL16 GFP CRL-2917™
NM2C5 CRL-2918™
NM2C5 GFP CRL-2919™

OLGA-PH-J/92 [OL-J/92] (ATCC® CRL-2576™) — This cell line was originally deposited as a crayfish cerebral ganglion cell line. However, cytochrome c oxidase subunit I (COI) testing at ATCC cannot confirm the crayfish origin. Based on this analysis, the distribution of ATCC® CRL-2576™ has been discontinued.

EPC (ATCC® CRL-2872™) — Cytochrome c oxidase subunit I (COI) testing at ATCC revealed that EPC, originally deposited as a Carp cell line (Cyprinus carpio), was in fact a Fathead Minnow cell line (Pimephales promelas). Since the time of deposit, isoenzymology testing has correctly and consistently identified EPC (ATCC® CRL-2872™) as a fish cell line. However, isoenzymology does not allow for speciation between genus, and information regarding the species of fish was previously only provided by the depositor. These observations were confirmed via COI testing of the original stock available to ATCC.

MPanc-96 (ATCC® CRL-2380™) – STR analysis at ATCC revealed that MPanc-96, a human pancreatic cell line, has an STR pattern identical to that of another human pancreatic cell line, AsPC-1 (ATCC® CRL-1682™). These observations were confirmed with the original stock available to ATCC. The distribution of ATCC® CRL-2380™ has been discontinued.

SNB-19 (ATCC® CRL-2219™) and U-373 MG (ATCC® HTB-17™) – STR analysis at ATCC revealed that SNB-19, a human glioblastoma cell line has a STR pattern identical to that for U-373 MG (ATCC® HTB-17™). SNB-19 and U-373 MG also share derivative chromosomes. These observations were confirmed with the original stock available to ATCC. Since then distribution of SNB-19 was discontinued.

U-373 MG (ATCC® HTB-17™) – As a result of sequencing, the authenticity of ATCC® HTB-17™ has been questioned by R.F. Petersson in Stockholm and collaborator E.G. Van Meir in Atlanta (personal communication and see Ishii, N., et al. Brain Pathol 9: 469-79, 1999). They report similarities between U-373 MG (ATCC® HTB-17™) and another glioblastoma, U-251. The cell line U-373 MG, obtained from the original lab in Uppsala has differing genetic properties from the ATCC® HTB-17™ (U-373 MG). Following further investigations, ATCC stopped distribution of this cell line.

U-118 MG (ATCC® HTB-15™) AND U-138 MG (ATCC® HTB-16™) – Two human glioblastoma cell lines reportedly from different individuals have identical variable number of tandem repeats (VNTR) and similar STR patterns and karyotype. U-118 MG and U-138 MG are very similar cytogenetically and share at least six derivative marker chromosomes. ATCC will continue distribution with an alert that the two lines have similar origin.

ECV-304, a presumptive endothelial cell line (ATCC® CRL-1998™) and human bladder cell line T24 (ATCC® HTB-4™) – STR analysis at ATCC suggested that ECV-304 was a derivative of T24. In addition, karyotype analysis of the two cell lines showed that they shared-marker chromosomes. Combined, these results confirm that ECV-304 is a derivative of T24, a line that was developed years earlier.

It is important to emphasize that all stocks of ECV show similar properties, not just those from ATCC. It is clear that the contamination took place before the cell banks independently received initial stocks. ATCC has written to each recipient of ATCC® CRL-1998™ (ECV-304) to inform them of these findings.

If additional misidentified cell lines are discovered ATCC scientists will report these general findings on this page after the depositors and other cell resource banks have been informed.