Authentication of scientific human cell lines: easy-to-use DNA fingerprinting.
Human cell lines are an important resource for research and most often used in reverse genetic approaches or as in vitro model systems of human diseases. In this regard, it is crucial that the cells faithfully correspond to the purported objects of study. A number of recent publications have shown an unacceptable level of cell lines to be false, in part as a result of the nonavailability of a simple and easy DNA profiling technique. We have validated different single- and multiple-locus variable numbers of tandem repeats (VNTRs) enabling the establishment of a noncommercial, but good laboratory practice, method for authentication of cell lines by DNA fingerprinting. Polymerase chain reaction amplification fragment length polymorphism (AmpFLP) of six prominent and highly polymorphic minisatellite VNTR loci, requiring only a thermal cycler and an electrophoretic system, was proven as the most reliable tool. Furthermore, the generated banding pattern and the determination of gender allows for verifying the authenticity of a given human cell line by simple agarose gel electrophoresis. The combination of rapidly generated DNA profiles based on single-locus VNTR loci and information on banding patterns of cell lines of interest by official cell banks (detailed information at the website www.dsmz.de) constitute a low-cost but highly reliable and robust method, enabling every researcher using human cell lines to easily verify cell line identity.
Dirks
Wilhelm G
WG
DSMZ-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany.
Drexler
Hans G
HG
D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000737:chemistry / Q000235:genetics; D016172:DNA Fingerprinting; D017931:DNA Primers; D004587:Electrophoresis, Agar Gel; D006801:Humans; D018598:Minisatellite Repeats; D008970:Molecular Weight; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D004247:DNA