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Abstract
The use of misidentified cell lines contaminated by other cell lines and/or microorganisms has generated much confusion in the scientific literature. Detailed characterization of such contaminations is therefore crucial to avoid misinterpretation and ensure robustness and reproducibility of research. Here we use DNA-seq data produced in our lab to first confirm that the Hep2 (clone 2B) cell line (Sigma-Aldrich catalog number: 85011412-1VL) is indistinguishable from the HeLa cell line by mapping integrations of the human papillomavirus 18 (HPV18) at their expected loci on chromosome 8. We then show that the cell line is also contaminated by a xenotropic murine leukemia virus (XMLV) that is nearly identical to the mouse Bxv1 provirus and we characterize one Bxv1 provirus, located in the second intron of the pseudouridylate synthase 1 (PUS1) gene. Using an RNA-seq dataset, we confirm the high expression of the E6 and E7 HPV18 oncogenes, show that the entire Bxv1 genome is moderately expressed, and retrieve a Bxv1 splicing event favouring expression of the env gene. Hep2 (clone 2B) is the fourth human cell line so far known to be contaminated by the Bxv1 XMLV. This contamination has to be taken into account when using the cell line in future experiments.
Highlights
Continuous cell lines are a cornerstone of cellular and molecular biology as well as of biomedical research
One of the best characterized cases of xenotropic murine leukemia viruses (MLV) (XMLV) contamination is that of the so-called xenotropic murine leukemia virus-related virus (XMRV), which was originally thought to be a possible cause of prostate cancer and chronic fatigue s yndrome[26]
These results confirm that Hep[2] was contaminated by HeLa at some point during its propagation and they show that human papillomavirus 18 (HPV18) sequences integrated in this subclone of HeLa have the same characteristics as in other subclones in terms of position in the genome and expression p attern[6,7,34]
Summary
Continuous cell lines are a cornerstone of cellular and molecular biology as well as of biomedical research. In addition to standard short tandem repeat (STR) typing, known virus-HeLa cell junctions of HPV18 integrations have been used to unveil cases of cancer cell lines contamination by HeLa7 Multiple microorganisms such as bacteria, fungi, viruses or yeasts have been identified as persistent contaminants of widely-used cell lines[8,9,10]. Other well-characterized cases of XMLV contamination involve a virus originating from expression of a mouse chromosome 1 proviral locus called the B10 xenotropic virus 1 (Bxv1)[28] This provirus is present in many severe combined immunodeficient (SCID) and nude mice s trains[20] and Bxv[1] infection of human cell lines passaged onto these mice strains has been experimentally demonstrated[24]. Bxv[1] contamination has been detected in two prostate cancer cell lines, one B-cell line and two pancreatic β cell lines, in which it was shown to produce infectious viral particles[16,17,29]
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