Establishment And Application Of A New Technology For Human Cell Line Authentication

Cell line was a common material which was used in life science,clinical medicine field,its importance was self-evident.Cell line authenticationhas been neglected by many research.Misidentified and cross-contaminated cell lines are unfortunately commonplace.Until recently,many major journals and research agencies recommended that cell line should be verified before authenticity be-fore publication or inclusion in the application.Although there were many new method for cell line authentication,DNA profiling based on the short tandom repeat was still propose as the golden stardard method.Short tandom repeat wasone STR locus per 6-10 kb in human genome.Because of itswide distri-bution,large amount of information,high polymorphism and following Mendel’s genetic law,it was easy to PCR amplification and profiling,STR technology has been widely used in paternity testing and detection of cell line cross－contamination and cell line authentication.In this study,we present a comprehensive investigation of cross－contamination for a panel of 278 cancer cell lines from28 institutesin China us ing 21 loci shorttandem repeat profiling method.The results showed that a total of46%(128 of 278)cross-contamination were uncovered among the 278 cell line.Notable false cell lines(eg:Hep2 andEJ)were still used under their false i dentity.Strikingly,73%(52 of71)of lines established by the Chinese researchers were misidentified and accounted for nearly half of the misidentification(52of 128,40%).Further,67%(35 of 52)of the contaminants in cell lines established in laboratories of china were HeLa cells or a possible hybrid of HeLa with another cell line.Further-more,the bile duct cancer cell line HCCC-9810 and degenerative lung cancer Calu-6 exhibited 88.9%match in the ATCC database(9-loci),indicating that they were from the same origin.However,whenwe used 21-loci to compare these two cell lines with the same algorithm,the percent matchwas only 48.2%,indicating that these two cell lines were different.It was proved that the new STRtechnology with 21 loci could be used to identify human cell linesmore ac curately and reliablely.150 cell lines with unique profilesruled out cell line cross-contamination and confirmed the identity.It was of great practical significance to pro-vide a comparative standard for the STR profiling of human cell lines by constructing the STR database of cell lines with independent intellectual property rights.