Species identification and confirmation of human and animal cell lines: a PCR-based method.
Misidentification and cross-contamination of cell lines are major problems of cell cultures that can make scientific results and their reproducibility unreliable. This paper describes a PCR-based method for easily identifying or confirming the species of origin of cell lines by using a panel of oligonucleotides specific for the nine animal species most common in cell culture laboratories. A panel of 35 human and animal cell lines, whose species of origin were previously confirmed by isoenzyme assay, was studied with nine species-specific primer pairs that specifically anneal to DNA sequences codifying for human, cat, dog, mouse, rat, horse, rabbit, African Green monkey cytochrome c oxidase subunit I (cox I), and one primer pair specific for the cytochrome b gene of Chinese hamster. The amplified fragments were analyzed by electrophoresis in ethidium bromide-stained 2% agarose gels. The method is simple, rapid, highly sensitive, and useful for routinely monitoring the species identity of cell cultures.
Parodi
B
B
National Institute for Cancer Research, Genoa, Italy. iclc@isge.it
Aresu
O
O
Bini
D
D
Lorenzini
R
R
Schena
F
F
Visconti
P
P
Cesaro
M
M
Ferrera
D
D
Andreotti
V
V
Ruzzon
T
T
D000818:Animals; D002415:Cats; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D004285:Dogs; D006736:Horses; D006801:Humans; D007527:Isoenzymes / Q000235:genetics; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*; D011817:Rabbits; D051381:Rats; D012680:Sensitivity and Specificity; D013045:Species Specificity
D007527:Isoenzymes