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13 October 2007 Species identification in cell culture: a two-pronged molecular approach
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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.

Jason K. Cooper, Greg Sykes, Steve King, Karin Cottrill, Natalia V. Ivanova, Robert Hanner, and Pranvera Ikonomi "Species identification in cell culture: a two-pronged molecular approach," In Vitro Cellular & Developmental Biology - Animal 43(10), 344-351, (13 October 2007).
Received: 30 August 2007; Accepted: 10 September 2007; Published: 13 October 2007

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