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

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 interlaboratory study was an international effort that consisted of 12 participating laboratories representing institutions from academia, industry, biological resource centers, and government. 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 Consortium laboratory members, 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 labs agreeing on allele calls among the discordant samples was above 92%. 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 18 of the 19 mouse STR markers is capable of discriminating at the intraspecies level between mouse cell lines. Further studies are ongoing to refine the assay including (1) development of an allelic ladder for improving the accuracy of allele calling and (2) integration of stutter filters to identify true stutter.

Authentication, characterization and contamination detection of cell lines, xenografts and organoids by barcode deep NGS sequencing

“…Among these technologies, STR profiling has been most widely used and there is an STR standard (ASN-0002) for guiding human cell line authentication ( 20 ). A panel of 19 STR markers for mouse cell lines was also developed ( 21 ). The sensitivity of STR assays for detecting contaminants is ∼5–10% ( 8 ).…”

Targeting 14-3-3ζ Overcomes Resistance to Epidermal Growth Factor Receptor-Tyrosine Kinase Inhibitors in Lung Adenocarcinoma via BMP2/Smad/ID1 Signaling

“…The cells were maintained at 37°C in a humidified incubator with 5% CO 2 . Cell lines were periodically authenticated by short-tandem repeat (STR) profiling using previously reported methods ( 34 ).…”

CLASTR: The Cellosaurus STR similarity search tool ‐ A precious help for cell line authentication

“…It should also be noted that there have been some efforts recently to develop sets of STR markers for nonhuman species, specifically for dog and mouse …”

“…It should also be noted that there have been some efforts recently to develop sets of STR markers for nonhuman species, specifically for dog 9,10 and mouse. 11,12 Results CLASTR, the Cellosaurus STR similarity search tool provides an intuitive and reliable platform to perform similarity searches on the human STR profiles contained in the Cellosaurus resource. It allows the efficient identification of cell lines and thus helps to detect contaminated and/or misidentified samples.…”