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Cell Line Contamination

Mycoplasma Prevention and Detection

9/20/2018

Mycoplasma are a distinct group of bacterial strains notorious for their ability to contaminate cell cultures without resulting in media turbidity or other obvious symptoms. Mycoplasma infection can deprive host cells of nutrients and induce global changes in host gene expression, which in turn can affect data reproducibility and lead to the misinterpretation of experimental results. Best practices in cell culture coupled with routine testing is critical for the prevention and early detection of mycoplasma contamination. In this presentation, we discuss the history of mycoplasma contamination with a focus on current prevention and detection methods. We will also expand on the products and services offered by ATCC for routine mycoplasma testing, highlighting our new PCR-based mycoplasma detection service. 

Key Points

  • Mycoplasma contamination significantly affects cell line physiology, resulting in erroneous data and misinterpretation of results.
  • Laboratory best practices and routine testing are essential for minimizing mycoplasma contamination. 
  • ATCC provides all three of the common methods of mycoplasma detection, including direct culture, Hoechst DNA staining, and PCR-based testing.

CRISPR/Cas9 genome-editing technology is a powerful new tool for developing cell-based models with disease-relevant mutations. In this presentation, we describe how this advanced technology was used to introduce specific point mutation that confers drug resistance into the A375 melanoma line to create the MEK1 Q56P Mutant-A375 Isogenic Cell Line. The new isogenic cell line is resistant to both MEK and BRAF inhibitors and, like the current isogenic KRAS and NRAS Mutant-A375 Cell Lines, sensitive to combination therapies targeting both upstream and downstream elements of the Ras/Raf-MAPK signaling pathway, making it an ideal model for screening and evaluating novel therapeutics and combination treatments targeting multidrug-resistant melanomas.

Abstract:

Throughout the world, acute respiratory infections are responsible for millions of deaths annually and are a leading cause of mortality in children. Even if not fatal, these infections can be debilitating and often result in hospitalization. Viruses such as influenza virus, respiratory syncytial virus, and enterovirus are major contributors to this disease burden. Emerging viruses have also posed significant risks to human health, as demonstrated by the SARS coronavirus outbreak in 2003 and the recent emergence of the MERS coronavirus. This underscores the importance of effective respiratory virus diagnostic assays, vaccines, and antiviral therapies. In this presentation, we will summarize the impact of respiratory viral disease on global health initiatives; the latest efforts to control and prevent infections (e.g., the universal influenza vaccine); and the importance of authenticated viruses and derivatives in the development of diagnostic assays, vaccines, and antiviral therapeutics.

Key Points:

  • Endemic and emerging respiratory virus infections represent a significant global heath burden.
  • Understanding respiratory virus epidemiology and pathogenesis is critical for the development of antiviral therapies.
  • Authenticated viral standards and derivatives are essential tools for the development and validation of novel preventative and therapeutic techniques.