Authentication of Scientific Human Cell Lines: Easy-to-Use DNA Fingerprinting

Dirks, Wilhelm G.; Drexler, Hans G.

doi:10.1385/1-59259-838-2:035

Cited by 13 publications

(17 citation statements)

References 4 publications

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“…To circumvent this limitation, the DSMZ performs additionally cytogenetic analyses and uses an alternative genotyping technique, which is not controlled by the MMR system. 22 The MCA technique, described in our report, may represent a promising alternative or complementary method for the authentication of cell lines. In contrast to STR profiling that is affected by MMR defects, defective MMR did not negatively influence MCA results in our study.…”

Section: Discussionmentioning

confidence: 89%

“…In these cell lines, STR profiling may lead to erroneous results when the numbers of tetranucleotides in the analysed STR loci are altered by a selective bottleneck, e. g. isolation of subclones or single clones upon transfection. To circumvent this limitation, the DSMZ performs additionally cytogenetic analyses and uses an alternative genotyping technique, which is not controlled by the MMR system 22 . The MCA technique, described in this report, may represent a promising alternative or complementary method for the authentication of cell lines.…”

Section: Discussionmentioning

confidence: 99%

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High‐throughput SNP‐based authentication of human cell lines

Castro

Dirks

Fähnrich

et al. 2012

Intl Journal of Cancer

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168

147

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Use of false cell lines remains a major problem in biological research. Short tandem repeat (STR) profiling represents the gold standard technique for cell line authentication. However, mismatch repair (MMR) deficient cell lines are characterized by microsatellite instability, which could force allelic drifts in combination with a selective outgrowth of otherwise persisting side lines, and thus, are likely to be misclassified by STR-profiling. Based on the high-throughput Luminex platform, we developed a 24-plex SNP-profiling assay, called Multiplex Cell Authentication (MCA), for determining authentication of human cell lines. MCA was evaluated by analysing a collection of 436 human cell lines from the DSMZ, previously characterised by eight loci STR profiling. Both assays showed a very high degree of concordance and similar average matching probabilities (~1 × 10−8 for STR-profiling and ~1 × 10−9 for MCA). MCA enabled the detection of less than 3% contaminating human cells. Analysing MMR deficient cell lines, evidence was obtained for a higher robustness of the MCA compared to STR profiling. In conclusion, MCA could complement routine cell line authentication and replace the standard authentication STR technique in case of MSI cell lines.

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Section: Discussionmentioning

confidence: 89%

Section: Discussionmentioning

confidence: 99%

High‐throughput SNP‐based authentication of human cell lines

Castro

Dirks

Fähnrich

et al. 2012

Intl Journal of Cancer

Self Cite

168

147

View full text Add to dashboard Cite

show abstract

“…This combination allows discrimination of one biospecimen from another at the level of 10 6 . 10,11 This specificity is in our view sufficient for biobanking authentication QC purposes. Therefore, this is a rapid but highly reliable and robust method, enabling easy verification of biospecimen identity.…”

Section: Resultsmentioning

confidence: 92%

“…For that procedure, the PCR amplification of six minisatellite VNTR loci (Apo-B1: with expected product length 522-909 bp; Col2A1: 600-850 bp; D17S5: 168-1080 bp; D1S80: 400-940 bp; D2S44: 600-> 5000 bp; PAH: 370-760 bp), and one microsatellite locus (GTG N : 10-> 15000 bp) was performed. 10,11 After size determination of the amplicons from each loci on an agarose FIG. 1. A relationship between the examined samples.…”

Section: Dna Fingerprintingmentioning

confidence: 99%

DNA Fingerprinting: A Quality Control Case Study for Human Biospecimen Authentication

Kofanova

Mathieson

Thomas

et al. 2014

Biopreservation and Biobanking

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This case study illustrates the usefulness of the DNA fingerprinting method in biobank quality control (QC) procedures and emphasizes the need for detailed and accurate record keeping during processing of biological samples. It also underlines the value of independent third-party assessment to identify points at which errors are most likely to have occurred when unexpected results are obtained from biospecimens.

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“…The results of the cytogenetic analysis of the cell line did not reflect those of the metastatic tumor, suggesting that the cell line has arisen by clonal selection. To rule out mislabelling or contamination as possible explanations for this finding, 24 we carried out DNA fingerprinting analysis which confirmed the identity of the PER‐568 cell line and primary tumor tissue.…”

mentioning

confidence: 99%

Classic and desmoplastic medulloblastoma: Complete case reports and characterizations of two new cell lines

et al. 2009

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Medulloblastoma (MB) is the most common type of brain tumor affecting children. These tumors are a significant cause of childhood mortality and morbidity, and more effective and less invasive treatment options are urgently required. To achieve these aims, it will be critical to develop a more comprehensive understanding of the molecular pathogenesis of MB. At present, there are relatively few well-characterized MB cell lines available to the research community for the study of MB molecular and cellular biology. Here we present the case reports of two children diagnosed with classic and desmoplastic MB, and describe the characteristics of two new MB cell lines derived from these individuals. A number of genes encoding components of the sonic hedgehog (SHH) and WNT pathways were up-regulated in the desmoplastic relative to the classic MB cell line consistent with aberrant activation of these pathways in desmoplastic MB. These cell lines represent an additional resource for the analysis of diverse aspects of MB biology.

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Authentication of Scientific Human Cell Lines: Easy-to-Use DNA Fingerprinting

Cited by 13 publications

References 4 publications

High‐throughput SNP‐based authentication of human cell lines

High‐throughput SNP‐based authentication of human cell lines

DNA Fingerprinting: A Quality Control Case Study for Human Biospecimen Authentication

Classic and desmoplastic medulloblastoma: Complete case reports and characterizations of two new cell lines

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