Abstract
Establishment of continuous cell lines from human normal and tumor tissues is an extended and useful methodology for molecular characterization of cancer pathophysiology and drug development in research laboratories. The exchange of these cell lines between different labs is a common practice that can compromise assays reliability due to contamination with microorganism such as mycoplasma or cells from different flasks that compromise experiment reproducibility and reliability. Great proportions of cell lines are contaminated with mycoplasma and/or are replaced by cells derived for a different origin during processing or distribution process. The scientific community has underestimated this problem and thousand of research experiment has been done with cell lines that are incorrectly identified and wrong scientific conclusions have been published. Regular contamination and authentication tests are necessary in order to avoid negative consequences of widespread misidentified and contaminated cell lines. Cell banks generate, store and distribute cell lines for research, being mandatory a consistent and continuous quality program. Methods implementation for guaranteeing both, the absence of mycoplasma and authentication in the supplied cell lines, has been performed in the Andalusian Health System Biobank. Specifically, precise results were obtained using real time PCR detection for mycoplasma and 10 STRs identification by capillary electrophoresis for cell line authentication. Advantages and disadvantages of these protocols are discussed.
Highlights
The use of cultured cells that acquired the ability to proliferate indefinitely is an extended tool in research laboratories
Twenty-four supernatant samples from tumor cell lines generated by the Biobank were used to mycoplasma detection. 100 ll supernatants were heated at 95 °C for 10 min and centrifuged at 1000 g for 5 s to discard cellular debris
By conventional PCR a 481 bp band was visualized for internal control detection in negative and positive samples and a specific 260 ± 8 bp band for mycoplasma positive samples (Fig. 1a)
Summary
The use of cultured cells that acquired the ability to proliferate indefinitely is an extended tool in research laboratories. Cell lines are used as in vitro models of health and disease by retaining many of the properties of the parental tissue or cell type, including diseasespecific changes (Christine Alston-Roberts et al 2010; Shannon et al 2016). Because of those reasons, cell lines based screening platforms are excellent models to test new therapeutic approaches. Many cell lines currently used are contaminated with mycoplasma and/or are replaced by cells derived for a different origin without researcher knowledge (CapesDavis et al 2010; Drexler et al 2017). The consequences of widespread misidentified and contaminated cell lines are immeasurable and it cannot be ignored by the scientific community (Huang et al 2017)
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