First searchable database for DNA profiles of human cell lines: Sequentialuse of fingerprint techniques for authentication

The authenticity and freedom from cross-contaminants of a cell line are imp ortant prerequisites for any research, development or production programs i nvolving cell lines. Mini- and microsatellites in the human genome harborin g variable-numbers of tandem repeat (VNTR) DNA markers allow individualizat ion at the DNA level and are of practical value for genetic linkage mapping , forensic legal medicine, paternity testing, monitoring of bone marrow tra nsplants, and individualization of established cell lines. We have validate d fingerprint techniques of different single- and multiple-locus VNTRs enab ling the establishment of a searchable database of DNA profiles. As a resul t, multiplexed polymerase chain reaction amplification fragment length poly morphism (AmpFLP) of four prominent and highly polymorphic minisatellite VN TR loci was proven as the best tool for screening the uniqueness of DNA pro files in a fingerprint database. In order to avoid false positivity, identi cal or similar DNA profiles based on AmpFLP VNTR were tested further using a multi-locus fingerprint system. Our data demonstrate that misidentificati on remains a chronic problem among human continuous cell lines (detailed in formation at URL http://www.dsmz.de). The combination of rapidly generated DNA profiles based on single-locus VNTR loci, their authentication by scree ning the fingerprint database, and confirmation of duplicate banding patter ns using multilocus fingerprints constitute a highly reliable and robust me thod, which enables high fidelity and quality of maintenance independent fr om the quantity of individual cell lines.