DNA-FINGERPRINTING OF HUMAN CELL-LINES USING PCR AMPLIFICATION OF FRAGMENT LENGTH POLYMORPHISMS

Methods for monitoring cell line identification and authentication inc lude species-specific immunofluorescence, isoenzyme phenotyping, chrom osome analysis, and DNA fingerprinting. Most previous studies of DNA f ingerprinting of cell lines have used restriction fragment length poly morphism analysis. In this study, we examined the utility of an altern ative and simpler method of cell line DNA fingerprinting-polymerase ch ain reaction (PCR) amplification of fragment length polymorphisms. Fou rteen human cell lines previously found by other methods to be either related or disparate were subjected to DNA fingerprinting by PCR ampli fication of selected fragment length polymorphism loci. Cell identific ation patterns by this method were concordant with those obtained by i soenzyme phenotyping and restriction fragment length polymorphism-DNA fingerprinting, and were reproducible within and between assays on dif ferent DNA extracts of the same cell line. High precision was achieved with electrophoretic separation of amplified DNA products an high res olution agarose or polyacrylamide gels, and with fragment length polym orphism (FLP) loci-specific ''allelic ladders'' to identify individual FLP alleles. Determination of the composite fingerprint of a cell lin e at six appropriately chosen fragment length polymorphism loci should achieve a minimum discrimination power of 0.999. The ability of PCR-b ased fragment length polymorphism DNA fingerprinting to precisely and accurately identify the alleles of different human cell lines at multi ple polymorphic fragment length polymorphism loci demonstrates the fea sibility of developing a cell line DNA fingerprint reference database as a powerful additional tool for future cell line identification anti authentication.