CELL LINE INDIVIDUALIZATION BY STR MULTIPLEX SYSTEM IN THE CELL BANK FOUND CROSS-CONTAMINATION BETWEEN ECV304 AND EJ-1/T24

Abstract

Short tandem repeat (STR) regions represent highly polymorphic microsatellite markers in the human gnome that have tandemly repetitive sequence elements of 2 to 7 by in length as a unit. The application of STR regions to population genetics and personal identification has been well studied. Recent technical advances have enabled us to analyze multilocus STR regions simultaneously by a method, called the STR Multiplex system, that uses a single PCR amplification in one tube. We established a new evaluation system for the identification of cell lines based on an STR Multiplex method that uses 9 different loci: D5S818, D13S317, D7S820, D16S539, vWA, TH01, Amelogenin, TPDX, and CSF1P0. The STR profiling data from 96 cell lines were examined and an efficiency of this approach for cell standardization was found. Using this method, we have analyzed the STR profiles of human cell lines, ECV304, EJ1, and 124, recently reported by the DSMZ-German Collection of Microorganisms and Cell Cultures to have been cross-contaminated. Our results clearly detect the cross-contamination between ECV304 and EJ-1/T24. The cross-contamination was estimated to be derived from the T24 cells. Collectively, the STR Multiplex system provides a rapid, precise, and powerful method in cell line identification for quality control at the JCRB Cell Bank.