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Deoxyribonucleic Acid Profiling Analysis of 40 Human Thyroid Cancer Cell Lines Reveals Cross-Contamination Resulting in Cell Line Redundancy and Misidentification

Division of Endocrinology, Diabetes and Metabolism (R.E.S., J.P.K., U.P., B.R.H.), Department of Medicine and University of Colorado Cancer Center DNA Sequencing and Analysis Core (C.K.), University of Colorado Cancer Center, University of Colorado Denver, Aurora, Colorado 80045; Human Oncology and Pathogenesis Program (M.B., J.A.K., J.A.F.), Memorial Sloan-Kettering Cancer Center, New York, New York, 10021; and Division of Endocrinology (R.C.S.), Internal Medicine Department, Department of Cancer Biology (L.A.M., J.A.C.), Mayo Clinic Comprehensive Cancer Center, Jacksonville, Florida 32224

Address all correspondence and requests for reprints to: Rebecca E. Schweppe, Ph.D., Division of Endocrinology, Diabetes and Metabolism, University of Colorado Denver, Aurora, Colorado 80045. E-mail: Rebecca.Schweppe{at}ucdenver.edu.

Context: Cell lines derived from human cancers provide critical tools to study disease mechanisms and develop novel therapies. Recent reports indicate that up to 36% of cell lines are cross- contaminated.

Objective: We evaluated 40 reported thyroid cancer-derived cell lines using short tandem repeat and single nucleotide polymorphism array analysis.

Results: Only 23 of 40 cell lines tested have unique genetic profiles. The following groups of cell lines are likely derivatives of the same cell line: BHP5-16, BHP17-10, BHP14-9, and NPA87; BHP2-7, BHP10-3, BHP7-13, and TPC1; KAT5, KAT10, KAT4, KAT7, KAT50, KAK1, ARO81-1, and MRO87-1; and K1 and K2. The unique cell lines include BCPAP, KTC1, TT2609-C02, FTC133, ML1, WRO82-1, 8505C, SW1736, Cal-62, T235, T238, Uhth-104, ACT-1, HTh74, KAT18, TTA1, FRO81-2, HTh7, C643, BHT101, and KTC-2. The misidentified cell lines included the DRO90-1, which matched the melanoma-derived cell line, A-375. The ARO81-1 and its derivatives matched the HT-29 colon cancer cell line, and the NPA87 and its derivatives matched the M14/MDA-MB-435S melanoma cell line. TTF-1 and Pax-8 mRNA levels were determined in the unique cell lines.

Conclusions: Many of these human cell lines have been widely used in the thyroid cancer field for the past 20 yr and are not only redundant, but not of thyroid origin. These results emphasize the importance of cell line integrity, and provide the short tandem repeat profiles for a panel of thyroid cancer cell lines that can be used as a reference for comparison of cell lines from other laboratories.

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