Webinars

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

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

Time: 12:00 PM Eastern Standard Time

Abstract:

Over the past few decades, much has been written about misidentification of cell lines, primarily human cell lines. It is not unusual that studies of the ‘same’ cell lines performed by different laboratories often show different results using the same methodology. Thus far, numerous cell lines are known to be misidentified due, in part, to lack of adequate testing. The financial loss incurred by using misidentified cell lines is estimated in the millions of dollars. Cell line authentication is crucial. The validity of scientific data demands that consistent and unequivocal verification of cell line identity is precise. The implementation of best tissue culture practices to include routine authentication of human cell lines by short tandem repeat (STR) profiling is highly recommended by funding agencies, journal editors, and scientific societies. This presentation will focus on the use of STR Profiling for human cell line authentication.

Key Points:

• Misidentification of cell lines may result in invalidated research results, withdrawal of funding, publication retraction, and loss of reputation
• Authentication of cell lines is critical to trusting your data
• STR profiling is an inexpensive, reliable method to authenticate human cell lines

Time: 12:00 PM Eastern Standard Time

Abstract:

The recent development of the CRISPR/Cas9 system provides a revolutionary gene-editing technology for basic research in biology and for development of targeted cancer therapies. In addition to enabling the identification of novel drug targets through functional screening, CRISPR/Cas9 facilitates the creation of disease models for drug discovery and development.

In this webinar, ATCC experts will address how ATCC utilized this advanced technology to create novel human cell models that contain disease-relevant point mutations and gene rearrangements. In addition, we will introduce a new type of BRAF inhibitor-resistant cell line that was created by using CRISPR/Cas9 to insert the NRAS Q61K mutation. These human isogenic lines provide useful disease models for the identification and validation of new therapeutics.

Key Points:

• CRISPR/Cas9 gene editing technology is a powerful tool for drug discovery
• Gene editing technology can be used to create disease-relevant cell models for screening new anti-cancer drug targets
• CRISPR/Cas9 is a useful tool for creating new types of drug-resistant cell models

Time: 12:00 PM Eastern Standard Time

Abstract:

Recent updates in the nomenclature rules for fungi (Melbourne Code, 2012) and improvements in contemporary molecular techniques have significantly affected fungal systematics and has triggered numerous taxonomic revisions among yeasts and fungi species in recent years. To deliver accurate and up-to-date information to our customers, we diligently characterize ATCC mycology products not only through rigorous quality control procedures, but also by various research projects. In this webinar, we will discuss how to identify fungi effectively and accurately under the current nomenclatural system by introducing a few recent studies on medically and industrially relevant yeasts and fungi.

Key Points:

• The new ‘One fungus, One name’ rule has significantly affected fungal systematics
• The quality control procedures used to evaluate and characterize ATCC mycology products meet the high standard of industrial demands
• Recent studies support MALDI-TOF mass spectrometry as a powerful rapid identification tool for yeasts and fungi

Time: 12:00 PM Eastern Standard Time

Abstract:

Assessing cell viability and cell health is crucial to all aspects of cell culture. In addition to the proper maintenance of cell stocks, cell health practices increase the level of confidence in experimental results. In this webinar presentation, we will discuss reliable and economic methods for determining cell proliferation and cell viability. Other cell culture practices and issues that affect cell health will also be addressed. These topics include choices in cell culture media/reagents, cryopreservation factors that affect cell lines and primary cells, and hard-to-detect mycoplasma contamination.

Key Points:

• Cell viability/proliferation can be measured quickly and easily with a number of assays, including the ATCC MTT and XTT proliferation kits, as well as the new Reliablue™ cell viability regent.
• When culturing specialty cells, such as stem cells or primary cells, certain considerations with choice of media and reagents must be taken.
• Mycoplasma contamination can cause chronic physiological problems with cultured cells and are hard to detect visually, but can be easily assessed with Mycoplasma Detection Kits and/or services, provided by ATCC.