Université Paris-Saclay, CNRS, IRD, UMR Évolution, Génomes, Comportement et Écologie, 91198, Gif-sur-Yvette, France.
Université de Poitiers, CNRS UMR 7267 Laboratoire Ecologie et Biologie des Interactions, Equipe Ecologie Evolution Symbiose, 5 Rue Albert Turpain, TSA 51106, 86073, Poitiers Cedex 9, France.
Sci Rep. 2020 Sep 29;10(1):16046. doi: 10.1038/s41598-020-73169-y.
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.
使用被其他细胞系和/或微生物污染的错误鉴定的细胞系在科学文献中造成了很多混淆。因此，详细地表征这种污染对于避免误解以及确保研究的稳健性和可重复性至关重要。在这里，我们使用我们实验室生成的 DNA-seq 数据首先证实 Hep2（克隆 2B）细胞系（Sigma-Aldrich 目录编号：85011412-1VL）通过在其预期的 8 号染色体位置上对人乳头瘤病毒 18（HPV18）的整合进行作图，与 HeLa 细胞系无法区分。然后，我们表明该细胞系还受到了类似于小鼠 Bxv1 前病毒的异嗜性鼠白血病病毒（XMLV）的污染，并且我们对位于假尿嘧啶核苷合成酶 1（PUS1）基因第二个内含子中的一个 Bxv1 前病毒进行了特征描述。使用 RNA-seq 数据集，我们证实了 HPV18 致癌基因 E6 和 E7 的高表达，表明整个 Bxv1 基因组中度表达，并检索到有利于 env 基因表达的 Bxv1 剪接事件。Hep2（克隆 2B）是迄今为止已知的第四个被 Bxv1 XMLV 污染的人类细胞系。在将来的实验中使用该细胞系时，必须考虑到这种污染。
