Molecular detection of cell line cross-contaminations using amplified fragment length polymorphism DNA fingerprinting technology |
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Authors: | E Milanesi P Ajmone-Marsan E Bignotti M N Losio J Bernardi F Chegdani M Soncini M Ferrari |
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Institution: | (1) Istituto di Zootecnica, Università Cattolica del Sacro Cuore, Via Emilia Parmense, 84, 29100 Piacenza, Italy;(2) Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna, 25124 Brescia, Italy |
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Abstract: | Summary We have tested amplified fragment length polymorphism (AFLP) technology, in comparison with isoenzyme analysis, for the simultaneous
detection of inter-and intraspecific cell line cross-contaminations (CCCs) in the cell line collection held at the Istituto
Zooprofilattico della Lombardia e dell’Emilia Romagna. Isoenzyme analysis identified four cases of interspecific CCCs. In
a single expreiment, AFLP was able to identify the species of origin of all cell lines for which a reference genomic deoxyribonucleic
acid was available and to detect five interspecific contaminations. Four CCCs confirmed data on isoenzymes, whereas the fifth
CCC was detected in a species for which isoenzyme analysis was noninformative. In addition, AFLP was able to identify the
putative source of the contaminations detected. The utility of the technology in the detection of intraspecific cell line
contaminations, depends on the number of cell lines that have to be distinguished in a specific species and on the availability
of highly informative fingerprinting systems. In mice, a single AFLP primer pair produced 16 polymorphisms and distinguished
all the 15 strains of mouse cell lines analyzed. In humans, 18 AFLPs identified 83 different profiles in the 159 cell lines
analyzed. Amplified fragment length polymorphism can conveniently be applied for cell line fingerprinting in species for which
hypervariable markers are not available. In species for which a highly informative multiplex of microsatellite markers is
available, AFLP can still provide a useful and cheap tool for simultaneously testing inter-and intraspecific contaminations. |
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Keywords: | cell cultures species origin isoenzymes molecular markers |
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