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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.

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.

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.

Keeping Cells Happy

Topics in Cell Health Maintenance and Viability

9/13/2018

Good practices are vital for safeguarding the health of cells in long-term culture. Cell health, maintenance, and viability cover a wide range of topics, from ensuring that cells are healthy in culture to having confidence that the cells are characterized correctly. This presentation will focus on topics such as cell viability assays that measure cell growth, authentication of cell lines through short tandem repeat (STR) profiling, mycoplasma testing, aseptic technique, and cryopreservation. From services to techniques, we will discuss ATCC’s role in ensuring the health and maintenance of cells in culture.

5/31/2018

Abstract:

Two of the major challenges that many scientists experience when developing a cell-based assay include obtaining cells with high biological relevance and then producing or procuring enough cells to run the assay without introducing cell variability. hTERT-immortalized primary cells address both issues. These cells are genetically modified such that the cells exhibit the growth characteristics of a continuous cell line but maintain the physiology of a primary cell. In this webinar, ATCC scientists will discuss our broad portfolio of hTERT-immortalized primary cells and provide some application data to illustrate how these cell models can easily be incorporated into your workflow. Special emphasis will be placed upon our new kidney transporter models for predictive toxicology (RPTEC/TERT1 OAT1, RPTEC/TERT1 OCT2, and RPTEC/TERT1 OAT3).

Key Points:

  • There is a lack of in vitro models that durably and correctly recapitulate in vivo physiology
  • hTERT-immortalized primary cells solve the problem of limited biological relevancy in cell-based assays
  • hTERT-immortalized primary cells exhibit the growth characteristics of a continuous cell line but maintain the physiology of a primary cell
  • ATCC has created kidney cell models using a well-characterized hTERT-immortalized RPTEC that stably overexpress the OAT1, OCT2, or OAT3 gene; our data show that these modified cell lines are very useful tools that provide kidney tissue-relevant results, improved consistency over time, and predictability for clinical trials

Cells utilize networks that span both temporal and spatial organizations, encompassing many individual steps of regulation. While the regulatory regimes to build networks in synthetic biology has grown from solely transcription to also include protein or RNA modalities, circuits comprised solely of protein-protein interactions have yet to be produced. Here, I'll describe several mechanisms relying on phosphorylation-activated localization and effector actuation for building OR and NOT gates from protein-protein phosphorylation events and their subsequent composition to form fast acting networks for ultrasensitive chemical sensing and phenotypic cellular control. Design and optimization of these networks were enabled by the use of a modular assembly method for rapid construction and testing of network variants. The final protein network spanned 15 individual member species to form a toggle switch that could sense chemical inputs as low as 1.0s in duration and maintain state over cellular division events. Motivated by these synthetic network designs, I will then describe one avenue in which synthetic biology network results can elucidate natural biological networks, i.e., synthetic biology-inspired discovery.

Abstract:

The species of the Mycobacterium tuberculosis Complex (MTBC)—M. tuberculosis, M. africanum, M. bovis, M. caprae, M. microti, and M. pinnipedii—are very closely related. In this webinar, we will discuss the techniques used to examine the MTBC in order to unravel this taxonomic mystery. Using phylogenomic techniques to compare the type strains of these species, we discovered that all of these “species” are, in fact, M. tuberculosis. We further examined all the strains deposited in GenBank under those species names and found all of them to be strains of M. tuberculosis. All known strains of three other putative MTBC members (“M. canettii”, “M. mungi”, and “M. orygis”) were similarly shown to be strain of M. tuberculosis. We have recently published a paper in the International Journal of Systematic and Evolutionary Microbiology officially unifying the previously separate MTBC species as M. tuberculosis.

Key Points:

  • Using whole-genome sequencing (WGS) and phylogenomic analysis of the MTBC species type strains, we discovered that all of these “species” are, in fact, Mycobacterium tuberculosis
  • By similarly analyzing all the MTBC non-type strain whole-genome sequences (>3,700) in GenBank, we determined that all of these strains similarly should be considered to be strains of Mycobacterium tuberculosis
  • We recommend the use of the infrasubspecific term ‘variant’ and infrasubspecific designations that generally retain the historical nomenclature associated with the groups or otherwise convey such characteristics (e.g., M. tuberculosis variant bovis). 
  • ATCC is currently in the process of updating the nomenclature used in our catalog to reflect this phylogenomically modernized taxonomy.

Abstract:

Optimized cell lines are essential for modeling neurodegenerative diseases such as Parkinson’s disease, screening novel therapeutics for preclinical studies, and testing the neurotoxicity of environmental compounds. Neural progenitor cells (NPCs) derived from induced pluripotent stem cells (iPSCs) are excellent in vitro models as they can be induced to differentiate down all three neural lineages. ATCC has recently added a line of NPCs developed from a donor with Parkinson’s disease to its collection of neurological research tools. This webinar will describe how ATCC NPCs can be differentiated into three neural fates and used in toxicological studies, focusing on the performance of the Parkinson’s disease-derived NPCs.

Key Points:

  • ATCC has a wide range of whole-cell models of Parkinson’s disease
  • NPCs cultured in ATCC Dopaminergic Neuron Differentiation Media express tyrosine hydroxylase and TuJ1
  • Parkinson’s disease patient-derived NPCs can be induced to differentiate into all three major neural lineages
  • Parkinson’s disease patient-derived NPCs can be used to screen compounds for toxicity

Abstract:

Kidney membrane transporters are key to drug disposition and renal clearance. Primary renal proximal tubule epithelial cells (RPTEC) are the most physiologically relevant cell models, but lose OAT1 and OCT2 expression in culture. Primary RPTEC transiently expressing these transporters show large variations between production lots. Furthermore, cell line-based models either do not have the kidney tissue origination or are tumor-derived. This presentation will introduce transporter cell models using hTERT-immortalized RPTEC that stably overexpress the OAT1 or OCT2 gene. Our data show that these cell lines provide tissue-relevant results, improved consistency over time, and predictability for clinical trials.

Key Points:

  • There is a lack of in vitro models that durably and correctly recapitulate kidney physiology
  • ATCC has created kidney cell models using hTERT-immortalized RPTEC that stably overexpress the OAT1 or OCT2 gene
  • hTERT-immortalized RPTEC provide kidney tissue-related results, improved consistency over time, and predictability for clinical trials

10/12/2017

Time: 12:00 PM Eastern Standard Time

Abstract:

Employing good aseptic technique and understanding the specific culture requirements of a strain are essential for optimal viral growth. In this webinar, we will discuss the basic principles for propagating animal viruses in the laboratory. Here, we will highlight best practices for making good virus stocks with an emphasis on the art and science of virus culture that should be followed to help avoid contamination and cross-contamination. Further, we will provide an in-depth look at viral culturing practices in both cell culture and embryonated chicken eggs using Influenza virus as an example.

Key Points:

  • Good aseptic technique alone is not sufficient to grow and maintain virus stocks
  • Host cell culture authentication is important for growing a good stock of virus
  • The nature of biomaterials used in culturing viruses in cells and the tendency for viruses to mutate are the primary challenges in obtaining consistent end results