Cell lines authentication and mycoplasma detection as minimun quality control of cell lines in biobanking

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.

Keywords: Biobanking; Cell line authentication; Mycoplasma; PCR; Quality control; STRs.

Fig. 1

Representative negative and positive results for mycoplasma detection. a Conventional PCR + Bioanalyzer, b Real-time PCR

Fig. 2

Results obtained using 5 STRs loci detection method. Four cell lines were compared with the original tissue (A and A′, B and B′, C and C′, D and D′), 3 cell lines were compared with frozen blood from the original donor (E and E′, F and F′, G and G′), and 3 cell lines were compared with FTA punch (H and H′, I and I′, J and J′)