Identification and authentication of animal cell culture by polymerase chain reaction amplification and DNA sequencing.
Polymerase chain reaction (PCR) amplification and deoxyribonucleic acid (DNA) sequence analysis were used to identify the species origin of cell lines used in a cell culture facility where various cell lines of different species are routinely propagated. The aldolase gene family was selected for PCR amplification because the DNA sequences of this gene are highly conserved over a wide range of animals and humans. A total of 36 cell lines representing 13 different species were selected for this study. The DNA from each cell line was amplified, and PCR products were analyzed by agarose gel electrophoresis. The results showed unique profiles of amplified bands on agarose gels that allowed differentiation among non-closely related species. However, DNA amplification of closely related species, including rat and mouse or human and primate, resulted in similar and indistinguishable banding patterns that could be further differentiated by DNA sequence analysis. These results suggested that aldolase gene amplification coupled with DNA sequence analysis is a useful tool for identification of cell lines and has potential application for use in identification of interspecies cross-contamination.
Liu
Merry Y
MY
Biologics Branch, Scientific Resources Program, National Center for Infectious Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road, MS-D43, Atlanta, Georgia 30333, USA. mkl6@cdc.gov
Lin
Seh-Ching
SC
Liu
Hsi
H
Candal
Francisco
F
Vafai
Abbas
A
D000818:Animals; D001483:Base Sequence; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D017931:DNA Primers; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics; D005784:Gene Amplification; D000882:Haplorhini; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D051381:Rats; D015183:Restriction Mapping / Q000379:methods
D017931:DNA Primers; D005634:Fructose-Bisphosphate Aldolase