The percentage of CD133+ cells in human colorectal cancer cell lines is influenced by Mycoplasma hyorhinis infection

BMC Cancer

Research Article

Peppino Mirabelli¹  Giuliana Fortunato¹  Rosa Di Noto¹  Francesca D'Alessio¹  Elisabetta Mariotti¹  Luigi Del Vecchio¹  Marica Gemei² 

[1] CEINGE - Biotecnologie Avanzate, via Comunale Margherita 482, 80145, Napoli, Italy;Dipartimento di Biochimica e Biotecnologie Mediche, Università Federico II, via Pansini 5, 80131, Napoli, Italy;CEINGE - Biotecnologie Avanzate, via Comunale Margherita 482, 80145, Napoli, Italy;European School of Molecular Medicine, CEINGE - Biotecnologie Avanzate, via Comunale Margherita 482, 80145, Napoli, Italy;

关键词: Enrofloxacin;    Human Colon Cancer Cell Line;    Continuous Cell Line;    Mycoplasma Species;    Mycoplasma Detection;   

DOI  :  10.1186/1471-2407-10-120

 received in 2009-12-07, accepted in 2010-03-30,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundMollicutes contamination is recognized to be a critical issue for the cultivation of continuous cell lines. In this work we characterized the effect of Mycoplasma hyorhinis contamination on CD133 expression in human colon cancer cell lines.MethodsMycoAlert® and mycoplasma agar culture were used to detect mycoplasma contamination on GEO, SW480 and HT-29 cell lines. Restriction fragment length polymorphism assay was used to determine mycoplasma species. All cellular models were decontaminated by the use of a specific antibiotic panel (Enrofloxacin, Ciprofloxacin, BM Cyclin 1 and 2, Mycoplasma Removal Agent and MycoZap®). The percentage of CD133 positive cells was analyzed by flow cytometry on GEO, SW480 and HT-29 cell lines, before and after Mycoplasma hyorhinis eradication.ResultsMycoplasma hyorhinis infected colon cancer cell lines showed an increased percentage of CD133+ cells as compared to the same cell lines rendered mycoplasma-free by effective exposure to antibiotic treatment. The percentage of CD133 positive cells increased again when mycoplasma negative cells were re-infected by Mycoplasma hyorhinis.ConclusionsMycoplasma hyorhinis infection has an important role on the quality of cultured human colon cancer cell lines giving a false positive increase of cancer stem cells fraction characterized by CD133 expression. Possible explanations are (i) the direct involvement of Mycoplasma on CD133 expression or (ii) the selective pressure on a subpopulation of cells characterized by constitutive CD133 expression.In keeping with United Kingdom Coordinating Committee on Cancer Research (UKCCCR) guidelines, the present data indicate the mandatory prerequisite, for investigators involved in human colon cancer research area, of employing mycoplasma-free cell lines in order to avoid the production of non-reproducible or even false data.

【 授权许可】

Unknown   
© Mariotti et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.