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Cited by 108Identification and Quantification in FoodForensic and Genetic ResearchGenetic diversity and population structure

Investigation of Cross-Contamination and Misidentification of 278 Widely Used Tumor Cell Lines

Open access
Huang, Yaqing, Liu, Yuehong, Zheng, Congyi, Shen, Chao·2017·PLOS ONE
DOI 10.1371/journal.pone.0170384PMC PMC5249119
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Abstract

In recent years, biological research involving human cell lines has been rapidly developing in China. However, some of the cell lines are not authenticated before use. Therefore, misidentified and/or cross-contaminated cell lines are unfortunately commonplace. In this study, we present a comprehensive investigation of cross-contamination and misidentification for a panel of 278 cell lines from 28 institutes in China by using short tandem repeat profiling method. By comparing the DNA profiles with the cell bank databases of ATCC and DSMZ, a total of 46.0% (128/278) cases with cross-contamination/misidentification were uncovered coming from 22 institutes. Notably, 73.2% (52 out of 71) of the cell lines established by the Chinese researchers were misidentified and accounted for 40.6% of total misidentification (52/128). Further, 67.3% (35/52) of the misidentified cell lines established in laboratories of China were HeLa cells or a possible hybrid of HeLa with another kind of cell line. Furthermore, the bile duct cancer cell line HCCC-9810 and degenerative lung cancer Calu-6 exhibited 88.9% match in the ATCC database (9-loci), indicating that they were from the same origin. However, when we used 21-loci to compare these two cell lines with the same algorithm, the percent match was only 48.2%, indicating that these two cell lines were different. The SNP profiles of HCCC-9810 and Calu-6 also revealed that they were different cell lines. 150 cell lines with unique profiles demonstrated a wide range of in vitro phenotypes. This panel of 150 genomically validated cancer cell lines represents a valuable resource for the cancer research community and will advance our understanding of the disease by providing a standard reference for cell lines that can be used for biological as well as preclinical studies.

AI snapshot

Written from the paper’s full text.

TL;DR

STR profiling of 278 tumor cell lines from 28 institutes in China found that 46.0% (128/278) were cross-contaminated or misidentified, with Chinese-established lines especially affected (73.2%, 52/71). The study also shows that 21-loci STR plus SNP genotyping can distinguish HCCC-9810 and Calu-6, which 9-loci ATCC profiling falsely matched at 88.9%.

Key findings

  • A total of 128/278 (46.0%) cross-contamination/misidentification cases were uncovered from 22 institutes; 20/278 (7.2%) failed PCR and were confirmed non-human, complete analysis succeeded for 258/278 (92.8%), and 150 cell lines were unique.
  • Among cell lines established in China, 52/71 (73.2%) were misidentified, and 35/52 (67.3%) of those were HeLa or a possible HeLa hybrid; in the non-Chinese model, 64/193 (33.2%) were misidentified or cross-contaminated. Overall, HeLa caused 60/128 (46.9%) of cross-contamination, affecting 31 cell lines.
  • HCCC-9810 and Calu-6 showed 88.9% match with the ATCC 9-loci database, but only 48.2% match with 21-loci STR; 24-loci SNP genotyping found 8 mismatches, indicating different origins.
  • Genetic instability was observed: 69 HeLa cell lines showed one or more STR alterations versus the prototype; nine other lines (293, BxPC-3, HT-29, DLD-1, HCT-116, Jurkat, Raji, HeLa, 5637) shared 80–94.12% identity to original lines after multiple passages; 32 cell lines with microsatellite instability were identified.

Methods

  • The authenticity of 278 tumor cell lines deposited in CCTCC by 28 institutes was examined; DNA was extracted and profiled using the Microreader TM21D System, co-amplifying 20 STR loci plus amelogenin, then compared with ATCC and DSMZ databases using the ATCC algorithm (≥80% related, <55% not related, 55–80% requiring further authentication).
  • Sensitivity for intraspecies cross-contamination was tested with AsPC-1:HeLa mixtures at 99:1, 95:5, 90:10, and 80:20 ratios; species identification used 10 species-specific primer sets; HCCC-9810 and Calu-6 were further characterized by 24-loci SNP genotyping.

Limitations

  • Stated: current STR databases such as ATCC, DSMZ, and JCRB provide only 9-loci profiles; 9 markers did not reliably discriminate HCCC-9810 and Calu-6, and authentication improved with 21-loci STR and SNP genotyping; the paper calls for careful and frequent authentication, possibly by more than one method.
  • Evident: the sample consists of cell lines deposited in CCTCC from 28 institutes in China, so the rates may not generalize elsewhere; 20 non-human samples failed PCR and 14 cell lines lacked original background documents, limiting classification; some STR profiles were non-detectable; the text reports both 46.0% and 46.4% for the overall contamination rate.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1371/journal.pone.0170384

Cites

39 of 39 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

13 of 34 cited sources checked · 5 supported · 1 flagged · accuracy not yet measured

  • Premisecitation

    Normal diploid cell lines CCC-HIE-2, FHC and FSH-74int established by the Chinese Academy of Medical Sciences had been shown to be actually derived from HeLa cells.

    “Normal diploid cell lines, namely CCC-HIE-2, FHC and FSH-74int, established by the Chinese Academy of Medical Sciences [32, 33], respectively, had been shown to be actually derived from HeLa cells.”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [32] KM Siddiqui, 1984, Primary and long term epithelial cell cultures from human fetal normal colonic mucosa, In vitro source
    • cites [33] I Elisia, 2013, Modulation of NF-kappaB and Nrf2 control of inflammatory responses in FHs 74 Int cell line is tocopherol isoform-specific, American journal of physiology Gastrointestinal and liver physiology source

      The Materials and Methods (Cell culture; page unavailable) describes FHs 74 Int as a primary cell line derived from a human fetal small intestine, contrary to the claim that it was derived from HeLa cells. Our copy does not address HeLa derivation of CCC-HIE-2 or FHC. (LLM, conf=0.84)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
  • Premisecitation

    Two adenoid cystic carcinoma, ACC-2 and ACC-M were reported as being contaminated by HeLa.

    “Two adenoid cystic carcinoma, ACC-2 and ACC-M were reported as being contaminated by HeLa [22].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [22] J Phuchareon, 2009, Genetic profiling reveals cross-contamination and misidentification of 6 adenoid cystic carcinoma cell lines: ACC2, ACC3, ACCM, ACCNS, ACCS and CAC2, PloS one source
  • Interpretationcitation

    The 8 SNP mismatches suggested that HCCC-9810 and Calu-6 were from different origins.

    “The SNP genotyping at 24-loci showed that there were 8 mismatches loci between HCCC-9810 and Calu-6 cell line, suggesting that they were from different origins [30].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [30] F Castro, 2013, High-throughput SNP-based authentication of human cell lines, Int J Cancer source

      The Results section ('Probability of discrimination and pair-wise analysis'; page unavailable) says that 0–2 mismatches across 24 SNPs indicate identical cell lines, supporting the inference that eight mismatches suggest different origins. Our copy does not report an eight-mismatch comparison between HCCC-9810 and Calu-6; searches for both cell-line names found no matching passage. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
  • Findingcitation

    SNP genotyping at 24 loci showed 8 mismatches between HCCC-9810 and Calu-6.

    “The SNP genotyping at 24-loci showed that there were 8 mismatches loci between HCCC-9810 and Calu-6 cell line, suggesting that they were from different origins [30].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [30] F Castro, 2013, High-throughput SNP-based authentication of human cell lines, Int J Cancer source

      The Methods (MCA-SNP typing; Statistics, page unavailable) describe genotyping 24 SNP loci. The Results (Analysis of DSMZ cell line collection by MCA, page unavailable) confirm that all 24 loci could be typed simultaneously, but do not report a comparison of HCCC-9810 with Calu-6 or eight mismatches; searches for both cell-line names found no such result. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
  • Findingcitation

    WISH was on the panel of cell models for differentiation in these institutions under its false identity as an amnion cell line of human origin.

    “WISH, which has been reported as a HeLa contaminant for decades [37], was also on the panel of cell models for differentiation [38] in these institutions under its false identity-amnion cell line of human origin.”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [37] DA Kniss, 2014, Discovery of HeLa Cell Contamination in HES Cells: Call for Cell Line Authentication in Reproductive Biology Research, Reproductive sciences source

      The Introduction and Discussion (page numbers unavailable) identify WISH as an amnion-derived cell line contaminated with HeLa and report its use in reproductive-biology research. They do not say WISH was on a panel of models for differentiation at the institutions described in the citing sentence. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [38] XD Chen, 2012, Astragalus induces human amniotic epithelial cells (WISH) to differentiate toward neurons, inhibits the expression of Notch1 and promotes cell survival, Sheng li xue bao: [Acta physiologica Sinica]
  • May combine two claims. 23
  • Classified as probably not a substantial claim. 7

  • May combine two claims.

    Premisecitation

    The bladder cancer cell line T24 was established in 1972 and has contributed to several cell line cross-contamination.

    “The bladder cancer cell line T24 was established in 1972 and has contributed to several cell line cross-contamination [20, 21].”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [20] G Johnen, 2013, Cross-contamination of a UROtsa stock with T24 cells—molecular comparison of different cell lines and stocks, PloS one source

      The Discussion says T24 was established in 1973, not 1972. The Discussion and Introduction support the other part of the claim: they describe T24 as a frequent source of cell-line contamination and identify multiple affected lines. (LLM, conf=0.90)

      Experimental — accuracy not yet measured
    • cites [21] B Nabarra, 2005, Neoplastic transformation and angiogenesis in the thymus of transgenic mice expressing SV40 T and t antigen under an L-pyruvate kinase promoter (SV12 mice), International journal of experimental pathology source

      The Abstract and Materials and methods describe SV12 transgenic mice and thymic tumours, not the T24 bladder cancer cell line. Searches for T24, its establishment in 1972, and cell-line cross-contamination found no passage addressing either part of the claim. (LLM, conf=0.75)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Microsatellite instability (MSI) was reported in leukemia, gastric, ovarian, breast, and colon cancers.

    “Microsatellite instability (MSI) was reported in leukemia [25], gastric [26], ovarian [27], breast [28], and colon cancers [29].”
    Quote in the paperNot a substantial claim2 of 3 sources checked
    • cites [25] W Parson, 2005, Cancer cell line identification by short tandem repeat profiling: power and limitations, The FASEB journal source
    • cites [26] Y Yao, 2004, Alterations of DNA mismatch repair proteins and microsatellite instability levels in gastric cancer cell lines, Laboratory investigation; a journal of technical methods and pathology source
    • cites [27] S V, 2014, Microsatellite instability, promoter methylation and protein expression of the DNA mismatch repair genes in epithelial ovarian cancer, Genomics source

  • May combine two claims.

    Premisecitation

    CNE-2, derived from Chinese patients, was reported bearing HPV sequences and suspected to be contaminated by fusion of HeLa with another nasopharyngeal cell line.

    “CNE-2, derived from the Chinese patients, was reported bearing HPV sequences and suspected to be contaminated by fusion of HeLa with another nasopharyngeal cell line [23].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [23] SY Chan, 2008, Authentication of nasopharyngeal carcinoma tumor lines, International journal of cancer source

  • May combine two claims.

    Findingcitation

    Four examples in the non-Chinese model were notably false cell lines (U-251, Hep2, Ej and HBL-100) for which the actual identity had been previously reported, contributing 6.3% (4/64) of the contamination rate.

    “including four examples that were notably false cell lines (U-251, Hep2, Ej and HBL-100) for which the actual identity had been previously reported [8–12], contributing 6.3% (4/64) of the contamination rate.”
    Quote in the paperNot a substantial claim1 of 5 sources checked
    • cites [8] 2010, Cell line misidentification: the beginning of the end, Nature reviews Cancer source
    • cites [9] S Azari, 2007, Profiling and authentication of human cell lines using short tandem repeat (STR) loci: Report from the National Cell Bank of Iran, Biologicals: journal of the International Association of Biological Standardization source
    • cites [10] PL Lorenzi, 2009, DNA fingerprinting of the NCI-60 cell line panel, Molecular cancer therapeutics source

      The Introduction and Results and Discussion (Matching Profiles; page unavailable) report that SNB-19 is derived from U251, supporting a previously reported identity relationship involving U251. They do not identify U251 itself as a false cell line or address Hep2, Ej, HBL-100, the non-Chinese model, or the 6.3% (4/64) figure. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [11] JR Masters, 2001, Short tandem repeat profiling provides an international reference standard for human cell lines, Proceedings of the National Academy of Sciences of the United States of America source
    • cites [12] K Yoshino, 2006, Essential role for gene profiling analysis in the authentication of human cell lines, Human cell source

  • May combine two claims.

    Findingcitation

    The HaCaT, HUVEC, SV-HUC-1, and hEM15A cell lines were totally free from cross-contamination.

    “The HaCaT [34], HUVEC [35], SV-HUC-1[36], and hEM15A cell lines established from immortalized human skin cutis cells, umbilical cords, ureteral epithelium and endometrial stromal cells were totally free from cross-contamination.”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [34] P Boukamp, 1988, Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line, The Journal of cell biology source
    • cites [35] SE Hughes, 1996, Functional characterization of the spontaneously transformed human umbilical vein endothelial cell line ECV304: use in an in vitro model of angiogenesis, Experimental cell research source
    • cites [36] CA Reznikoff, 1988, Neoplastic transformation of SV40-immortalized human urinary tract epithelial cells by in vitro exposure to 3-methylcholanthrene, Carcinogenesis source

  • May combine two claims.

    Findingown data

    The subclones generated from K562 showed a high concordance of the clonotype homogeneity, i.e., one different clonotype was observed among the six K562 cell lines and the profile of the subclones differed from the parental culture by a single change of STR.

    “In sharp contrast, the subclones generated from K562 showed a high concordance of the clonotype homogeneity, i.e., one different clonotype was observed among the six K562 cell lines and the profile of the subclones differed from the parental culture by a single change of STR.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Different samples of SGC-7901, SMMC-7721, BEL-7402, BEL-7404, BEL-7704, HL-7702, BGC-823 and TSCCA cell lines collected from independent laboratories were contaminated by either HeLa or another contaminant, possibly caused by a second intruder when they were cultured.

    “Different samples of SGC-7901, SMMC-7721, BEL-7402, BEL-7404, BEL-7704, HL-7702, BGC-823 and TSCCA cell lines collected from independent laboratories were contaminated by either HeLa or another contaminant, possibly caused by a second intruder when they were cultured (Table 3).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    95 C was obtained from the primary tumor site while 95 D originated from lymph nodes of the metastatic site.

    “95 C was obtained from the primary tumor site while 95 D originated from lymph nodes of the metastatic site.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    When 21-loci were used to compare HCCC-9810 and Calu-6 with the same algorithm, the percent match was only 48.2%, indicating these two cell lines were different.

    “However, when we used 21-loci to compare these two cell lines with the same algorithm, the percent match was only 48.2%, indicating that these two cell lines were different.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The match algorithm and similarity thresholds used to compare STR profiles correctly indicate whether two cell lines are the same or different.

  • May combine two claims.

    Findingown data

    Two multilocus profiles revealed a close relationship between LNCaP and EJ, which was a vector for T24 cell line.

    “Two multilocus profiles revealed a close relationship between LNCaP and EJ, which was a vector for T24 cell line.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: The match algorithm and similarity thresholds used to compare STR profiles correctly indicate whether two cell lines are the same or different.

  • May combine two claims.

    Findingown data

    In the non-Chinese model, 129 instances out of 193 characterized samples had distinct STR profiles and consisted of 78 individual cell lines.

    “129 instances out of 193 characterized samples had distinct STR profiles and consisted of 78 individual cell lines.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The ATCC and DSMZ cell bank STR databases used for comparison are accurate and authoritative references for cell line identity.

  • May combine two claims.

    Findingown data

    Our findings showed evidence of 46.9% (60/128) cross-contamination was caused by HeLa, affecting 31 cell lines, which were purported to have been established from 10 types of tumor and 3 types of normal tissue.

    “Our findings showed evidence of 46.9% (60/128) cross-contamination was caused by HeLa, affecting 31 cell lines, which were purported to have been established from 10 types of tumor (colon, liver, breast, stomach, lung, nasopharynx, tougue, laryngeal, salivary gland and bronchia) and 3 types of normal tissue (lung, intestine and liver).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The subclones of these nine cell lines shared 80-94.12% identity to the original cell lines and appeared to bear null allele and mutations.

    “The subclones of these nine cell lines shared 80–94.12% identity to the original cell lines and appeared to bear null allele and mutations (Table 4).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    20 samples failed to be amplified by PCR and were confirmed to be inter-species cross-contamination.

    “84.4% (108/128) of cross-contamination was intra-species while 20 samples failed to be amplified by PCR and confirmed to be inter-species cross-contamination.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    A total of 32 cell lines with MSI were identified with three banding patterns, including 7 liver cancer cell lines, 8 cervical cancer cell lines, 2 ovarian cancer cell lines, 1 leukemia cell line, 3 stomach cancer cell lines, 4 breast cancer cell lines, 1 colon cancer cell line, 2 osteosarcoma cancer cell lines, 1 pancreatic cancer cell line, 1 kidney cell line, 1 lung cancer cell line and 1 lymph

    “A total of 32 cell lines with MSI were identified with three banding patterns, including 7 liver cancer cell lines, 8 cervical cancer cell lines, 2 ovarian cancer cell lines, 1 leukemia cell line, 3 stomach cancer cell lines, 4 breast cancer cell lines, 1 colon cancer cell line, 2 osteosarcoma cancer cell lines, 1 pancreatic cancer cell line, 1 kidney cell line, 1 lung cancer cell line and 1 lymph”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In the Chinese model, only 19 instances out of 71 had distinct STR profiles (26.8%) and consisted of 11 individual cell lines.

    “In the Chinese model, only 19 instances out of 71 had distinct STR profiles (26.8%) and consisted of 11 individual cell lines (Table 3).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The original descriptions or documents of cell lines accurately identify their tissue of origin and whether they were established in China.

  • May combine two claims.

    Findingown data

    vWA, D19S433, D16S539 and D13S317 loci were the least involved, showing three alleles at one locus.

    “On the other hand, vWA, D19S433, D16S539 and D13S317 loci were the least involved, showing three alleles at one locus.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingnone given

    CCTCC repository generated a new 21-loci STR database.

    “CCTCC repository generated a new 21-loci STR database (www.CCTCC.org/STR.html)).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Zos and Zos-M were established respectively from the primary tumor site and the skip metastasis of an osteosarcoma patient.

    “Similarly, Zos and Zos-M were established respectively from the primary tumor site and the skip metastasis of an osteosarcoma patient [14].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [14] C-y Zou, 2008, Establishment and characteristics of two syngeneic human osteosarcoma cell lines from primary tumor and skip metastases, Acta Pharmacologica Sinica source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Microsatellite instability, characterized by the occurrence of new alleles in tumors, is a major event in certain carcinomas brought about by mismatch repair deficiency.

    “In contrast to the loss of heterozygosity, microsatellite instability [24], characterized by the occurrence of new alleles in tumors, is a major event in certain carcinomas brought about by mismatch repair deficiency.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [24] H Yamamoto, 2015, Microsatellite instability: an update, Archives of toxicology source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Interpretationreasoning

    This discrepancy could be resulted from either a technical issue in PCR, in which one allele was preferentially amplified while another was below the detectable level, or a biological variation where the Jurkat cell line from one laboratory had lost one allele.

    “This discrepancy could be resulted from either a technical issue in PCR, in which one allele was preferentially amplified while another was below the detectable level, or a biological variation where the Jurkat cell line from one laboratory had lost one allele.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    HCCLM-3 was a tumorigenic subclone of MHCC-97L and MHCC-97H, and was established from nude mice.

    “HCCLM-3 was a tumorigenic subclone of MHCC-97L and MHCC-97H, and was established from nude mice [15].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [15] J Yang, 2005, Molecular cytogenetic characteristics of the human hepatocellular carcinoma cell line HCCLM3 with high metastatic potential: comparative genomic hybridization and multiplex fluorescence in situ hybridization, Cancer genetics and cytogenetics source

  • May combine two claims.

    Premisecitation

    The Y chromosome is unstable, and some cell lines and even normal cells from older male donors lost the Y chromosome, such as A549 and Jurkat.

    “Since Y chromosome was unstable, some cell lines and even normal cells from older male donors lost the Y chromosome [13], such as A549 and Jurkat cell line.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [13] HG Drexler, 2002, Bladder carcinoma cell line ECV304 is not a model system for endothelial cells, In Vitro Cellular & Developmental Biology-Animal source

  • May combine two claims.

    Premisecitation

    HeLa and its derivatives HeLa S3, AV3 and WISH contaminations are notorious problems and may be simply detected by either isoenzyme or STR profiling.

    “HeLa [16] and its derivatives HeLa S3, AV3 [17] and WISH contaminations are notorious problems and may be simply detected by either isoenzyme [18] or STR profiling [19].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [16] J Fogh, 1977, Absence of HeLa cell contamination in 169 cell lines derived from human tumors, Journal of the National Cancer Institute source
    • cites [17] JR Masters, 2002, HeLa cells 50 years on: the good, the bad and the ugly, Nature reviews Cancer source
    • cites [18] SM Gartler, 1968, Apparent Hela cell contamination of human heteroploid cell lines, Nature source

  • May combine two claims.

    Premisecitation

    Fang Ye and colleagues investigated 380 human cell lines and found about 25% were cross-contaminated or misidentified.

    “An alarming rate of cross-contamination among human cell lines used in China was reported by Fang Ye and colleagues [6] who investigated 380 human cell lines in which about 25% was cross-contaminated or misidentified.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [6] F Ye, 2015, Genetic profiling reveals an alarming rate of cross-contamination among human cell lines used in China, FASEB journal: official publication of the Federation of American Societies for Experimental Biology source

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Each sample was repeated at least twice to confirm the results.

    “Each sample was repeated at least twice to confirm the results.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Well-characterized and validated reference data obtained from ATCC and DSMZ were used for data comparison.

    “For data comparison, well-characterized and validated reference data obtained from ATCC and DSMZ were used.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S4 (based).

    Study design and conductnone given

    If a sample could not be amplified in three time repeats, it was considered as non-human origin.

    “If the sample could not be amplified in three time repeats, it will be considered as non-human origin.”
    Quote in the paperNot a substantial claim
28 of 28

  • The paper relies on this without support.

    The ATCC and DSMZ cell bank STR databases used for comparison are accurate and authoritative references for cell line identity.

    “By comparing the DNA profiles with the cell bank databases of ATCC and DSMZ”
    “had authenticated prototypes deposited in cell repositories or research institutions worldwide”

  • The paper relies on this without support.

    The original descriptions or documents of cell lines accurately identify their tissue of origin and whether they were established in China.

    “The verified cell lines were derived from 20 different tissues (Table 2).”

  • The paper relies on this without support.

    The 278 cell lines from 28 institutes in China are representative of widely used tumor cell lines in China.

    “a panel of 278 cell lines from 28 institutes in China”
    “278 samples collected from 28 independent sources of China are divided into 2 groups”

  • The paper relies on this without support.

    A unique STR profile is sufficient to consider a cell line genomically validated and suitable as a reference.

    “150 cell lines with unique profiles demonstrated a wide range of in vitro phenotypes.”
    “This panel of 150 genomically validated cancer cell lines represents a valuable resource”

  • The paper relies on this without support.

    The match algorithm and similarity thresholds used to compare STR profiles correctly indicate whether two cell lines are the same or different.

    “when we used 21-loci to compare these two cell lines with the same algorithm, the percent match was only 48.2%, indicating that these two cell lines were different.”
    “The pairwise comparison of STR profiles revealed three matched pairs with similarity scores >0.9.”

  • The paper relies on this without support.

    The misidentification and cross-contamination of cell lines are a widespread problem in research, dating back as far as 45 years ago.

    “The misidentification and cross-contamination of cell lines are deemed to be a widespread problem in research, dating back as far as 45 years ago.”

  • The paper relies on this without support.

    Spontaneous immortalization is rare in human cell lines.

    “Instructively, spontaneous immortalization is rare in human cell lines.”

  • The paper relies on this without support.

    Current STR databases such as ATCC, DSMZ and JCRB only provide 9-loci STR profiles.

    “Unfortunately, currant STR databases such as ATCC, DSMZ and JCRB only provide 9-loci STR profiles.”

  • The paper relies on this without support.

    Tainted cell lines are still on sale for use.

    “Despite sporadic reports about cell line cross-contamination provided by cell repositories in China through systematic quality control check of their own cell lines, the tainted cell lines are still on sale for use.”

  • The paper relies on this without support.

    The grave problem of cell line misidentification in China has been neglected.

    “The grave problem of cell line misidentification in China has been neglected.”

  • The paper relies on this without support.

    Lack of information has prevented researchers from keeping up to date with literature reports of cell line misidentification.

    “Lack of information has prevented researchers from keeping up to date with literature reports of cell line misidentification.”

  • The paper relies on this without support.

    The grave problem continues to fester due to the negligence of cell line users and originators as well as journal editors and funding agencies.

    “Although the international cell repositories have compiled a list of cross-contaminated cell lines to alert their users, the grave problem continues to fester due to the negligence of cell line users and originators as well as journal editors and funding agencies.”

  • The paper relies on this without support.

    STR-based profiling methods have been used for the authentication of cell lines for more than 24 years.

    “STR-based profiling methods have been used for the authentication of cell lines for more than 24 years.”

  • The paper relies on this without support.

    There are variations in the numbers of loci and in the specific loci comprising the published profiling.

    “However, there are variations in the numbers of loci and in the specific loci comprising the published profiling.”
14 of 14

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Some studies have shown that misidentification and mycoplasma contamination are still common issues.

    “Some studies have shown that misidentification and mycoplasma contamination are still common issues (Horbach and Halffman, 2017; Huang et al., 2017; Olarerin-George and Hogenesch, 2015; Timenetsky et al., 2006).”

    From Recommendations on fit-for-purpose criteria to establish quality management for microphysiological systems and for monitoring their reproducibility (2024) · cites this paper as Huang et al., 2017

    The Discussion (page unavailable in our copy) describes cell-line misidentification as widespread, and reports authentication problems in 46.4% of the cell lines studied. It recommends mycoplasma contamination tests but does not report that mycoplasma contamination is common; that part of the claim is unsupported by this source. (LLM, conf=0.91)

    Experimental — accuracy not yet measured

  • Levels of cell line contamination for China are between 25% and 46% according to recent publications.

    “several recent publications indicate levels of cell line contamination for China between 25% [13] and 46% [46]”

    From The ghosts of HeLa: How cell line misidentification contaminates the scientific literature (2017) · cites this paper as [46]

  • The CCCM rate among Chinese-derived cell lines was 73.2% (52/71).

    “Again, the CCCM rate among Chinese-derived cell lines was much higher: 73.2% (52/71) [104].”

    From The history, landscape, and outlook of human cell line authentication and security (2024) · cites this paper as [104]

  • A 2017 21-STR evaluation of 278 lines originating from 28 Chinese sources found a CCCM rate of 46%, and 22 of the sources had contributed contaminated lines.

    “Two years later, a 21-STR evaluation of 278 lines originating from 28 Chinese sources found a CCCM rate of 46% (128/278); 22 of the sources had contributed contaminated lines [104].”

    From The history, landscape, and outlook of human cell line authentication and security (2024) · cites this paper as [104]

  • China has been a member of the World Federation for Culture Collections since 1987 and has made bioeconomic development a strategic priority.

    “A member of the World Federation for Culture Collections (”

    From The history, landscape, and outlook of human cell line authentication and security (2024) · cites this paper as [104]

    The Results and Discussion address cell-line authentication and misidentification in China, not membership in the World Federation for Culture Collections. Searches for the federation, 1987, and bioeconomic development found no passage supporting either part of the claim in our copy. Page numbers were not provided. (LLM, conf=0.76)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • STR locus name differs between methods and results

    Minor

    The methods identify D16S1043 as one of the amplified loci, whereas the results tables label that locus D6S1043.

  • SNP assayed in Table 6 differs from the selected-SNP list

    Minor

    The methods list rs1800896 among the 24 selected SNPs, but the 24-locus results table reports rs1800869 instead. The text does not establish whether these are meant to be the same assay.

2 of 2

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1 of the 40 references couldn’t be matched to a known paper.

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Related

Authors
Huang, Y., Liu, Y., Zheng, C., & Shen, C.
Affiliations
Wuhan University (CN)
Corresponding author
  • Zheng, Congyi
  • Shen, Chao
ORCID
Venue
PLOS ONE
Publisher
Public Library of Science (PLoS)
Published
2017-01-20
Volume
12
Issue
1
Pages
e0170384
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0170384
PMC
PMC5249119
OpenAlex
W2574356023
Semantic Scholar
31b0e5a474894e03ce42b50eedbb3b34366a26db
Cited by
105
References
40
Citations by year
2026: 8 · 2025: 9 · 2024: 25 · 2023: 8 · 2022: 11 · 2021: 12 · 2020: 9 · 2019: 7 · 2018: 11 · 2017: 5
FWCI
3.58
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • Figshare (submittedVersion, cc-by)
Funders
  • National Natural Sciences Foundation of China (31370185)
  • National Basic Research Program of China (NSTI-CR14, NSTI-CR15)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Identification and Quantification in Food
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Forensic and Genetic Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic diversity and population structure
Keywords
cancer cell lines, cross-contamination, cell line misidentification, short tandem repeat profiling, cell line authentication, HeLa cells
MeSH terms
China, Humans, Neoplasms, Genomic Instability, Cell Line, Tumor

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-28.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-28.

Cited sources are checked by gpt-6-sol.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

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