We’ll start with a scary but true fact: As reported at Lab Manager in 2011 and other publications, in Europe,mycoplasma contamination levels have been found to be extremely high—between 25 percent and 40 percent—and reported rates in Japan have been as high as 80 percent. In the US this value is lower but above 15 percent due to testing availability. This number is, unfortunately, on the rise. In the last decade alone bacterial contamination incidence almost doubled. Mycoplasmas are extremely detrimental to any cell culture: they affect the host cells’ metabolism and morphology, cause chromosomal aberrations and damage, and can provoke cytopathic responses, rendering any data from contaminated cultures unreliable.
Out of all of the bacterial contaminants, mycoplasmas are the most serious and widespread – and the most detrimental, owing to their low detection rates and their toxicity to mammalian cells (Drexler and Uphoff, Cytotechnology 2002). While technically bacteria, mycoplasmas are smaller, have no visible appearance, and lack a cell wall, which makes them resistant to antibiotics and allows them to filter through sterilization media. In recent years, there has been a growing interest to understand and develop robust tools for rapid mycoplasma detection. Approaches for the detection of mycoplasmas traditionally employ cell culture (Dvorakova et al. Vet. Med. 2005), which makes for a laborious process in terms of time and resources. On the other hand, more recent strategies employ the polymerase chain reaction (PCR) to detect mycoplasmas, which gives results in a few hours. It is based on the amplification of DNA from cell culture supernatant via PCR which can then be visualized using gel electrophoresis. While guidelines for mycoplasma testing are addressed in several international pharmacopoeias (United States Pharmacopoeia, European Pharmacopoeia and Japanese Pharmacopoeia), PCR-based detection kits are only just finding regulatory acceptance – in 2007, the European Pharmacopoeia described foundational guidelines for the use of such kits for mycoplasma detection and the Japanese pharmacopoeia has published similar guidelines. While the United States Pharmacopoeia is yet to establish such rules, it does state that any alternative method for the detection of mycoplasma outside of the established cell culture-based approaches must prove to be comparable in terms of quality.
Because of our ever-growing commitment to controllable, safe and clean laboratory environments, Akron has developed a best-in-class range of mycoplasma solutions. From treatment solutions to detection kits, with Akron’s cleanroom products you can make contamination a thing of the past. The primer set – specific to a segment of the 16S rRNA region of the mycoplasma genome – enables the detection of the various species of mycoplasma, (including M. pneumoniae, M. fermentans, M. arginini, M. salivarium, M. hominis, M. pulmonis, M. arthritidis, M. bovis, M. pirum and more), with extremely high sensitivity. It gives off a 270bp product. To shave off even more detection time, Akron is releasing an RT-PCR mycoplasma kit, which enables simultaneous amplification and detection. Easy to use. Will be launching officially in October.
Because a good, clean culture doesn’t just happen – it has to start that way – Akron’s commitment to a contaminant-free, sustainable laboratory starts at the root – with the selection of raw materials. One of the most overlooked, yet most frequent sources of variability in mammalian cell culture is the composition of the raw materials. This is even more critical in biopharmaceuticals production settings, where regulatory requirements put extreme pressure on the quality of the final product. The European Medicines Agency and the European Pharmacopoeia have outlined a series of guidelines for cell therapy products to highlight specific control measures to be followed to ensure raw material quality. These are fundamental to the development of cell-based products and cover raw material origin, source and quality.
To learn more about the importance of raw materials in cell therapy and Akron’s commitment to a sustainable, clean and safe laboratory – and learn about our products that span the drug development pipeline – come and meet us at the International Society for Cellular Therapy’s North America Annual Meeting in Philadelphia from September 8-11, 2013. Akron will have a booth at the meeting and our staff will be happy to talk and answer any questions. In addition to that, Akron’s CEO Dr. Claudia Zylberberg will chair a session on Raw Materials and their impact on cell therapy. If you would like to schedule an appointment to discuss customized products or services, feel free to email us at firstname.lastname@example.org.