Interlaboratory study to validate a STR profiling method for intraspecies identification of mouse cell lines

Published

June 20, 2019

Author(s)

Jamie L. Almeida, Aleksandra Dakic, Karin Kindig, Maikan Kone, Deborah L. Letham, Scott Langdon, Ruth Peat, Jayamalini Holding-Pillai, Erin Hall, Mark Ladd, Megan Shaffer, Heath Berg, Jinliang Li, Georges Wigger, Steven P. Lund, Carolyn R. Steffen, Barbara Fransway, Bob Geraghty, Manuela Natoli, Beth Bauer, Susanne M. Gollin, Dale Lewis, Yvonne A. Reid

Abstract

The Consortium for Mouse Cell Line Authentication was formed to validate Short Tandem Repeat (STR) markers for intraspecies identification of mouse cell lines. The STR profiling method is a multiplex polymerase chain reaction (PCR) assay comprised of primers targeting 19 mouse STR markers and two human STR markers (for interspecies contamination screening). The goals of the Consortium were to perform an interlaboratory study to – (1) validate the mouse STR markers to uniquely identify mouse cell lines (intraspecies identification), (2) to provide a public database of mouse cell lines with the National Institute of Standards and Technology (NIST)- validated mouse STR profiles, and (3) to publish the results of the interlaboratory study. The study was based on 50 of the most commonly used mouse cell lines obtained from the American Type Culture Collection (ATCC). Of the 50 mouse cell lines, 18 had unique STR profiles that were 100% concordant (match) among all 12 laboratories, and the remaining 32 cell lines had discordance that was resolved readily and led to improvement of the assay. The discordance was due to low signal and interpretation issues involving artifacts and genotyping errors. Although the total number of discordant STR profiles was relatively high in this study, the percent of agreement in allele calls among the discordant samples was above 91%. The STR profiles, including electropherogram images, for NIST-validated mouse cell lines will be published on the NCBI BioSample Database (https://www.ncbi.nlm.nih.gov/biosample/). Overall, the interlaboratory study showed that the multiplex PCR method using 19 mouse STR markers is capable of discriminating at the intraspecies level between mouse cell lines.

Citation

Plos Biology

Volume

14

Issue

6

Pub Type

Journals

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DOI Link

Keywords

mouse cell line authentication, genotyping cell lines, short tandem repeat, STR, validation of mouse multiplex PCR assay, identity testing of mouse cell lines

Citation

Almeida, J. , Dakic, A. , Kindig, K. , Kone, M. , Letham, D. , Langdon, S. , Peat, R. , Holding-Pillai, J. , Hall, E. , Ladd, M. , Shaffer, M. , Berg, H. , Li, J. , Wigger, G. , Lund, S. , Steffen, C. , Fransway, B. , Geraghty, B. , Natoli, M. , Bauer, B. , Gollin, S. , Lewis, D. and Reid, Y. (2019), Interlaboratory study to validate a STR profiling method for intraspecies identification of mouse cell lines, Plos Biology, [online], https://doi.org/10.1371/journal.pone.0218412 (Accessed November 23, 2024)