Species identification in cell culture: a two-pronged molecular approach
- 13 October 2007
- journal article
- Published by Springer Science and Business Media LLC in In Vitro Cellular & Developmental Biology – Animal
- Vol. 43 (10), 344-351
- https://doi.org/10.1007/s11626-007-9060-2
Abstract
Species identification of cell lines and detection of cross-contamination are crucial for scientific research accuracy and reproducibility. Whereas short tandem repeat profiling offers a solution for a limited number of species, primarily human and mouse, the standard method for species identification of cell lines is enzyme polymorphism. Isoezymology, however, has its own drawbacks; it is cumbersome and the data interpretation is often difficult. Furthermore, the detection sensitivity for cross-contamination is low; it requires large amounts of the contaminant present and cross-contamination within closely related species may go undetected. In this paper, we describe a two-pronged molecular approach that addresses these issues by targeting the mitochondrial genome. First, we developed a multiplex PCR-based assay to rapidly identify the most common cell culture species and quickly detect cross-contaminations among these species. Second, for speciation and identification of a wider variety of cell lines, we amplified and sequenced a 648-bp region, often described as the “barcode region” by using a universal primer mix targeted at conserved sequences of the cytochrome C oxidase I gene (COI). This method was challenged with a panel of 67 cell lines from 45 diverse species. Implementation of these assays will accurately determine the species of cell lines and will reduce the problems of misidentification and cross-contamination that plague research efforts.Keywords
This publication has 36 references indexed in Scilit:
- Comprehensive DNA barcode coverage of North American birdsMolecular Ecology Notes, 2007
- Cases of Mistaken IdentityScience, 2007
- bold: The Barcode of Life Data System (http://www.barcodinglife.org)Molecular Ecology Notes, 2007
- Identifying Canadian mosquito species through DNA barcodesMedical and Veterinary Entomology, 2006
- The problems and promise of DNA barcodes for species diagnosis of primate biomaterialsPhilosophical Transactions Of The Royal Society B-Biological Sciences, 2005
- Biological identifications through DNA barcodesProceedings Of The Royal Society B-Biological Sciences, 2003
- DNA Banks for Endangered Animal SpeciesScience, 2000
- Autoactivation of Avian Urokinase-type Plasminogen Activator (uPA)Published by Elsevier BV ,1998
- Contamination of Hodgkin's disease cell culturesNature, 1981
- Characterisation of human cell lines and differentiation from HeLa by enzyme typingNature, 1976