Detection algorithm for the validation of human cell lines

Eltonsy, Nevine H.; Gabisi, Vivian A.; Li, Xuesong; Russe, K. Blair; Mills, Gordon B.; Stemke-Hale, Katherine

doi:10.1002/ijc.27533

Cited by 14 publications

(12 citation statements)

References 23 publications

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“…STR profiles were cross-checked with the ATCC database. HCT116 and RKO cell lines with ≥ 80% match with the ATCC online STR database are considered valid (28). STR profiles of FET, CBS, HCT116b and TENN cell lines have never been reported before.…”

Section: Methodsmentioning

confidence: 99%

R-Spondin1/LGR5 Activates TGFβ Signaling and Suppresses Colon Cancer Metastasis

et al. 2017

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Leucine rich repeat containing G protein-coupled receptor 5 (LGR5), an intestinal stem cell marker, is known to exhibit tumor suppressor activity in colon cancer, the mechanism of which is not understood. Here we show that R-spondin 1 (RSPO1)/LGR5 directly activates TGFβ signaling cooperatively with TGFβ type II receptor in colon cancer cells, enhancing TGFβ-mediated growth inhibition and stress-induced apoptosis. Knockdown of LGR5 attenuated downstream TGFβ signaling and increased cell proliferation, survival, and metastasis in an orthotopic model of colon cancer in vivo. Upon RSPO1 stimulation, LGR5 formed complexes with TGFβ receptors. Studies of patient specimens indicate that LGR5 expression was reduced in advanced stages and positively correlated with markers of TGFβ activation in colon cancer. Our study uncovers a novel crosstalk between LGR5 and TGFβ signaling in colon cancer and identifies LGR5 as a new modulator of TGFβ signaling able to suppress colon cancer metastasis.

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Section: Methodsmentioning

confidence: 99%

R-Spondin1/LGR5 Activates TGFβ Signaling and Suppresses Colon Cancer Metastasis

et al. 2017

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“…Match criteria generally incorporate an algorithm to compare two STR profiles 7, 16, 30–32. One STR profile is defined as the “questioned” profile (the sample being tested), while the other is a “reference” profile, ideally from the same donor.…”

mentioning

confidence: 99%

“…Match criteria generally incorporate an algorithm to compare two STR profiles. 7,16,[30][31][32] One STR profile is defined as the ''questioned'' profile (the sample being tested), while the other is a ''reference'' profile, ideally from the same donor. Where another sample from that donor is not available, the questioned STR profile should be compared to other samples from that laboratory and databases online.…”

mentioning

confidence: 99%

“…5,6 Various match algorithms have been proposed for comparison of DNA fingerprints, some requiring specialized bioinformatics expertise. 32,33 Although the more complex algorithms can yield a substantial amount of information on authenticity and instability, early validation of STR profiling has shown that simple match algorithms also work well to discriminate between related and unrelated samples. 7,30 The STR profiles made available by the cell banks generally included eight STR loci plus amelogenin for gender determination.…”

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confidence: 99%

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Match criteria for human cell line authentication: Where do we draw the line?

Capes‐Davis

Reid

Kline

et al. 2012

Intl Journal of Cancer

150

119

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Continuous human cell lines have been used extensively as models for biomedical research. In working with these cell lines, researchers are often unaware of the risk of cross-contamination and other causes of misidentification. To reduce this risk, there is a pressing need to authenticate cell lines, comparing the sample handled in the laboratory to a previously tested sample. The American Type Culture Collection Standards Development Organization Workgroup ASN-0002 has developed a Standard for human cell line authentication, recommending short tandem repeat (STR) profiling for authentication of human cell lines. However, there are known limitations to the technique when applied to cultured samples, including possible genetic drift with passage. In our study, a dataset of 2,279 STR profiles from four cell banks was used to assess the effectiveness of the match criteria recommended within the Standard. Of these 2,279 STR profiles, 1,157 were grouped into sets of related cell linesduplicate holdings, legitimately related samples or misidentified cell lines. Eight core STR loci plus amelogenin were used to unequivocally authenticate 98% of these related sets. Two simple match algorithms each clearly discriminated between related and unrelated samples, with separation between related samples at !80% match and unrelated samples at <50% match. A small degree of overlap was noted at 50-79% match, mostly from cell lines known to display variable STR profiles. These match criteria are recommended as a simple and effective way to interpret results from STR profiling of human cell lines.

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“…However, the problem still exists (Nardone 2008) and newer methods, like analysis of microsatellite short tandem repeats (STR) markers with high heterozygosity values, are being used to identify human and simian cells in place of older methods like isoenzyme profiling (Almeida et al 2011;Eltonsy et al 2012). The former requirement comes from the known history of mis-identified cells, particularly HeLa being deposited in repositories.…”

Section: Cell Identity and Cell Productivitymentioning

confidence: 99%