Mycoplasma and Misidentified Cell Lines One Day Workshop
Workshop Success!
Mycoplasma and Misidentified Cell Lines – The Insidious Threats to Research – Risks, Issues and Solutions.
The two major risks facing any practitioner of cell culture: cell line misidentification and the contamination of cell cultures with Mycoplasma were explored in a new one day workshop hosted at ECACC March 2011. Due to the success of the workshop, attracting delegates from academic and commercial organisations from across Europe, it promises to become an annual event.
ECACC Mycoplasma & Misidentified Cell Lines Proceedings 2011
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ECACC would like to thank all the presenters who included internal specialists from the HPA Culture Collectons and external speakers such as Professor Glyn Stacey and Lyn Healy from the UK National Stem Cell Bank (NSCB), Ross Hawkins from NIBSC and representatives from Lonza and Life Technologies.
The workshop was designed to provide delegates with a rounded and balanced understanding of the topics and be able to make informed judgements about testing solutions. Furthermore delegates could return to their labs with a pragmatic and practical knowledge set to effectively deal with Mycoplasma and authentication issues and help answer these questions:
- How can I be confident that the cell line I propose to use is correctly identified and true to its claimed provenance?
- How can I confirm the identity of my cell line(s)?
- Modern DNA profiling methods for the identification of cell lines are designed for human cell lines. How do I confirm the identity of animal cell lines?
- How should I ensure that my cell stocks are free from Mycoplasma contamination?
- What working practices and screening procedures should I use to keep my cell culture laboratory free from Mycoplasma?
- What do I do if I suspect my cells are cross contaminated or contaminated with Mycoplasma?
The day began witha key note introduction to Good Cell Culture Practice (GCCP) guidelines, the history of cell line authentication and misidentification and how the portfolio of cell line authentication techniques has evolved since the problem was first brought to light in the 1960s. Subsequently a focused appraisal of Short Tandem Repeat (STR) profiling of human cell lines was given, including how to select a set of primers and the issues surrounding comparison of profiles from laboratory to laboratory. Developments at ECACC were presented such as the recent introduction of DNA bar-coding to replace the use of isoenzyme analysis for species identification.
The emphasis then shifted to Mycoplasma beginning with a detailed review of the biology and pathology of the Mycoplasma genus from the perspective of a microbiologist. Following this, testing methods for Mycoplasma were covered; the classic methods of detection by Culture Isolation and Hoechst staining and strategies to be employed in the design and selection of PCR primers for Mycoplasma testing and the relative merits and pitfalls of using 'off the shelf' single round PCR kits. Snapshot presentations of novel Mycoplasma detection techniques from Lonza (MycoAlert™) and Life Technologies’ (Myco Seq™) were given. An extended lunch break included a tour of the ECACC Mycoplasma and Molecular Biology laboratories.
The requirement for biological standards in Mycoplasma testing and cell line authentication, particularly with regard to the manufacture of novel cell based therapeutics was discussed.
A specialist responsible for the culturing of newly acquired cell cultures gave a fascinating insight into her experiences of dealing with misidentified, cross contaminated and Mycoplasma infected cell lines. This included some thought provoking data from experiments that simulated accidental low dose inoculation of cells into typical cell culture reagents, the surprising longevity of these cells and their ability to be recovered and cultured from these reagents after weeks of cold storage.
The workshop closed with a summary of conclusions taken from the discussions. These conclusions are well worth reading and can be viewed as part of the detailed write up of the proceedings available to download:
ECACC Mycoplasma & Misidentified Cell Lines Workshop Proceedings 2011
