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Cell Culture Contamination Part 1

Jeanne Phillips, Ph.D.
Recent studies have shown that at least 20% of the animal cell cultures currently in use in the US are contaminated by either microorganisms or other cell lines! This three part live, on-line seminar series will review the scope of this major problem and examine some of its causes and techniques for avoiding it.  It will also explore some key, easy to employ strategies for preventing these losses by careful culture management.

This webinar will discuss good asceptic technique: developing a practical approach to asceptic technique; reducing day-to-day contamination problems in the lab; helpful hings for avoiding contamination.
Jan 25 2011
39 mins
Cell Culture Contamination Part 1
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  • An Introduction To Cell Culture Apr 30 2015 4:00 pm UTC 60 mins
    Mark Rothenberg, Ph.D.
    This webinar will introduce the history, theory, basic techniques, and potential pit-falls of mammalian cell culture. It is designed for students and new lab technicians, as well as bench scientists interested in updating their techniques or knowledge in the field.

    Topics to be discussed include:
    • History and practical theories of cell culture and its impact on today’s science
    • The requirements needed to set up a cell culture laboratory
    • Challenges when performing mammalian cell culture and how to overcome them

    About our Presenter:
    Dr. Mark Rothenberg graduated from Emory University with his Ph.D. in Cell and Developmental Biology. Over the past 25 years, Mark has held positions in both academia and industry where he has developed an expertise in the areas of assay development and cell culture. He currently holds the position of Manager Scientific Training and Education with Corning Life Sciences.
  • Tips and Techniques for Serum-free Expansion of hMSCs Recorded: Mar 26 2015 53 mins
    Brian Posey
    Abstract:
    There is a great interest in application of human mesenchymal stem cells (hMSCs) in cell therapy and tissue engineering due to their self-renewal, multi-lineage differentiation, immunomodulation, and trophic potential. One of the challenges faced in the clinical application of hMSCs is the need for efficient expansion of these cells in vitro without altering their capacity. Serum-free mammalian cell culture media, in particular, require optimization of the expansion protocols. Even subtle changes in routine handling can have a significant impact on the cells’ potential.

    This seminar will cover the variables that can influence the desired regenerative and differentiation properties including medium selection, vessel surface treatment, impact of the cell source, and seeding density. We will also discuss how users can select the correct conditions for optimized growth and functionality.

    Speaker Biography:
    Brian Posey is a Product Development Manager for cell culture media at Corning Life Sciences. Brian has over 10 years experience in cell biology and industrial scale cGMP manufacturing of both liquid and powder cell culture media. Since joining Corning in 2012, Brian has lead numerous innovative technology projects for the media business ranging from customer technology transfer for production scale-up to developing new serum-free media for industrial and stem cell lines.
  • A Novel “Thaw and Go” Cell-based Model to Study SLC Transporters Recorded: Feb 24 2015 59 mins
    Na Li, Ph.D.
    Abstract:
    As a key determinant of drug pharmacokinetics, transporter mediated drug-drug interaction has garnered significant attention from the pharmaceutical industry and regulatory authorities. Corning offers a comprehensive list of tools to support drug transporter studies and recently introduced Corning® TransportoCells™ products to support in vitro assessment of drug interaction with SLC transporters. This new model provides a convenient “thaw and go” cell-based model with robust activity and consistent performance. In this webinar, we will provide an overview of Corning TransportoCells products along with applications for in vitro-to-in vivo correlation. Validation data will also be presented for the newly available TransportoCells products, including OATP1A2, OATP2B1, PEPT1, PEPT2, and NTCP.

    Presenter Biography:
    Dr. Na Li received her B.S degree in Biology from Fudan University, Shanghai, China, and her Ph.D. in Pharmacology from Dartmouth Medical School, Hanover, NH. Her major research focus is on drug transporters, including interspecies differences in hepatobiliary transporters, transporter quantification, and in vitro-to-in vivo extrapolation of drug pharmacokinetics. At Corning, Dr. Li contributes her expertise in in vitro drug transporter technology and its application in drug ADME.
  • 3D Tissue Models for Analyzing Dermatologic and Respiratory Systems Recorded: Jan 28 2015 51 mins
    Yukari Tokuyama, Ph.D.
    Join us on January 28th for a special Corning-sponsored webinar presented by ATCC®.

    Abstract:
    The significance of 3D tissue modeling opens up new possibilities for the study of complex physiological processes in vitro. Advances in cell isolation, media development, substrates, and growth surfaces are leading to culture environments that provide better biological and functional properties than traditional 2D cell culture. These models may provide a more predictive analysis and result in a more streamlined process of drug discovery and development. In this webinar, we will discuss recent developments in 3D modeling using ATCC primary and hTERT immortalized cells with specialized Corning® permeable support culture systems in dermatologic and respiratory studies.

    Presenter Biography:
    Dr. Yukari Tokuyama is a Field Application Scientist at ATCC. Prior to this role, she led the Stem Cell Product Development group and focused on products for human induced pluripotent stem cells and lineage specific differentiation. She earned her Ph.D. in Cell and Molecular Biology from the College of Medicine at the University of Cincinnati, where she studied the mechanism of genomic instability in cancer. She completed her post-doctoral training at the Oregon Health & Science University, Oregon National Primate Research Center, with a research focus on human and non-human primate stem cell biology.
  • Factors to Consider When Choosing Cell-based Assays for Use with 3D Cultures Recorded: Dec 16 2014 60 mins
    Terry Riss, Ph.D.
    Join us for a special Corning sponsored webinar presented by Promega Corporation.

    Cells cultured in 3D model systems often acquire relatively large in vivo-like structures compared to the thickness of a 2D monolayer of cells grown on standard plastic plates. Multicellular 3D culture systems containing more than one cell type and exhibiting formation of a complex extracellular matrix represent a more physiologically relevant environment, yet provide a challenge for assay chemistries originally designed for measuring events from monolayers of cells. There is an unmet need for guidelines for design and verification of convenient and effective assays useful for larger 3D microtissues. Critical factors to consider for each model system and cell type include effective penetration of detection reagents and/or complete lysis of microtissue structures using combinations of detergent and physical disruption. We will present the approach used to verify performance of a bioluminescent ATP detection assay for measuring cell viability, a caspase assay for detecting apoptosis, and cell stress reporter assays to detect mechanisms leading to cytotoxicity. Recommendations for factors to consider when verifying performance of cell health assays on 3D culture models will be presented.

    Speaker Bio:

    Dr. Terry Riss started the Cell Biology program at Promega Corporation in 1990 and has since held several R&D and Project Management positions. Dr. Riss managed development of cell viability, cytotoxicity, apoptosis, and protease assay systems and also led efforts to identify and promote multiplexing of cell-based assays to determine the mechanism of cell death. Dr. Riss now serves as Senior Product Specialist, Cell Health involved in outreach educational training activities including validating assay systems applied to 3D cell culture models.
  • Tips and Techniques for Culturing and Scaling-Up Primary Cells Recorded: Nov 19 2014 59 mins
    Mark Rothenberg, Ph.D.
    Primary cells can more closely mimic an in vivo-like state and generate more physiologically relevant data than immortalized cells. But unlike immortalized cells, primary cells have complex nutritional needs and require optimized growth conditions. In this webinar, you will learn:
    •Proven techniques for isolating primary cells from tissues
    •The importance of choosing the correct dissociation method
    •Troubleshooting techniques for culturing healthier primary cells.
    •Other topics to be discussed include:
    -Choosing the optimal cell culture surface
    -Selecting the optimal cell culture media 
    -Choosing the correct vessel for scaling-up
  • In Vitro Models For Studying Angiogenesis Recorded: Oct 30 2014 58 mins
    Paula M. Flaherty
    Angiogenesis is the process by which a new blood supply is established from pre-existing blood vessels. It is initiated by degradation of vessel basement membrane, endothelial cell proliferation, invasion, and directional migration towards chemoattractants, tube formation, and finally the establishment of a new vasculature.

    We will discuss a portfolio of Corning products designed to investigate specific stages of angiogenesis through standardized and quantitative in vitro cell-based assays.

    •HUVEC-2 Endothelial Cells—Widely studied endothelial cells that have been pre-screened for responsiveness to VEGF, a prototypic stimulator of angiogenesis
    •ECMs and Chemoattractants—ECMs and key pro-angiogenic chemoattractants for optimal EC propagation, attachment, differentiation, and other key functions
    •Endothelial Cell Invasion and Migration Assays—An enabling platform technology consisting of Corning® FluoroBlok™ microporous permeable supports coated with our unique extracellular matrix proteins for quantitation of endothelial cell migration and invasion
    •Endothelia Cell Tube Formation Assay—A rapid assay system allowing direct screening of angiogenic compounds for their effects on endothelial cell tube formation


    Biography:

    Paula Flaherty is a Technology Manager at Corning Life Sciences. Her team develops strategy and products focused on the modulation of in vitro cell behavior using extracellular matrix, media, vessel design, and growth factors. Prior to joining Corning Life Sciences, Paula studied retinal degeneration at the Berman-Gund Laboratory, Harvard Medical School in Boston, MA. She received her bachelor’s degree in Microbiology from the State University of New York and is an In Vitro Cell Biology Fellow, W. Alton Jones Cell Science Center in Lake Placid, NY.
  • Tips and Tricks to Help Develop and Run Microplate-based Assays Recorded: Sep 25 2014 59 mins
    Mark Rothenberg, Ph.D.
    Are you relying on biochemical and cell based in vitro assays in your research? In this webinar, Dr. Mark Rothenberg will share “insider” tips and tricks for enhancing your biochemical and cell-based assay performance.
    Critical parameters for achieving success with these assays include understanding the assay dynamics, the instrumentation involved in reading the assay, and, in the case of cell-based assays, the environment in which the cells are grown impacts cellular physiology. Factors such as microplate geometry, density, surfaces, and instrumentation all play important roles in the success of the assay.
    Learn how to:
    •Select the correct microplate and its role in the success of the assay
    •Choose the correct tip for your liquid handling needs
    •Determine the correct instrumentation and settings to run your assay
  • A Beginner’s Guide to Scaling-Up Cells Recorded: Aug 26 2014 58 mins
    Dr. Katherine Strathearn
    In the past several decades, the usage of mammalian cell culture processes have allowed for better understanding of basic research and manufacturing of higher quality products compared to previous methods. Although much progress has been developed throughout the years, limitations and challenges during optimization of such processes in scale-up of mammalian cell cultures may be encountered.

    This webinar will provide a simple guide and review of novel cell culture vessels and surface technologies that have enabled researchers to improve suspension and adherent mammalian cell culture scale-up. Additionally, this webinar will cover certain parameters to consider when scaling-up cells. Selecting the correct vessel and cell culture conditions will increase throughput without increasing laboratory space, incubator space, or time.
  • Genetic Alteration Cell Panels: Tools for High Throughput In Vitro Screening Recorded: Aug 6 2014 52 mins
    Fang Tian, Ph.D. & David Randle, PhD
    Extensive genomic sequencing efforts in recent years have provided detailed profiles of the somatic gene mutations that occur in a wide range of human cancers. In order to facilitate basic and translational cancer research, ATCC has designed and validated a number of genetic alteration cell panels targeting the key molecular pathways identified in these studies. To demonstrate suitability of the panels for high throughput screening, the EGFR panel was selected for evaluation using Corning® Epic® Technology, a label-free platform that uses optical biosensors for high sensitivity biochemical and cell-based assays.
    Label-free dynamic mass redistribution (DMR) responses in whole cells provide phenotypic activity profiles which are readily amenable to evaluation of compound activity and pharmacology. DMR responses obtained using the EGFR cell panel showed that Epic Technology can be utilized to evaluate receptor responsiveness to ligands and successfully predict drug response. Furthermore, label-free phenotypic responses can provide profiles of cellular signaling pathways downstream of receptor activation that may identify alternative targets for drug screening in the cell panel. In summary, combining Epic Technology and the EGFR genetic alternation panel offers convenient tools to screen for ligands or biologics that directly target or affect EGFR receptor biology.
  • Partnering With Corning® Gentest℠ Contract Research For In Vitro ADME Testing Recorded: Jul 22 2014 60 mins
    David M. Stresser, Ph.D.
    In this webinar, you will learn why partnering with Corning® GentestSM Contract Research will enable your company to reach goals faster. You will also learn about our latest service offerings, including new CYP induction and SLC transporter assay services - all designed to align with regulatory agency guidance documents. These services continue to strengthen our in vitro drug-drug interaction testing portfolio with a deep focus on enzyme inhibition, induction, and transport.
  • Applications of Corning® FluoroBlok™ Permeable Supports Recorded: Jun 25 2014 57 mins
    Jeffrey Partridge
    In vitro cell migration and invasion assays are frequently used as model systems for studying the directed movement of cells towards a chemoattractant stimulus, or to determine how a particular drug, growth factor or extra cellular matrix coating affects that movement. The classic method used to analyze this movement, the Boyden chamber, can be time-consuming, labor-intensive, and subjective.
    An improved version of this technology was developed, Corning FluoroBlok. As cells migrate or invade through a fluorescence blocking membrane, they are detected using a bottom-reading microplate reader or inverted fluorescence microscope. Cells remaining in the upper chamber of the insert are shielded from detection, allowing for quantitation of cell numbers in this homogeneous assay system.
    Recommended uses of the FluoroBlok system as well as recent improvements will be reviewed in this seminar.
  • Corning® Matrigel® Matrix: Material Properties, Applications and Proper Usage Recorded: May 22 2014 61 mins
    Shabana Islam, PhD
    The extracellular matrix (ECM) is a complex material that serves to stabilize tissues, promote cell attachment, and modulate cell functionality by specifically interacting with cell surface receptors. This material has been shown to play structural, biochemical and mechanical roles in the normal growth and differentiation of various cell types. Corning Matrigel Matrix, an ECM-based material isolated from EHS mouse tumors, is a widely used solubilized basement membrane preparation. Based on its physiological composition and functionality, Matrigel Matrix effectively models the physical interplay between diverse cell types and the ECM microenvironment in vivo.
  • Animal-free, Synthetic Surfaces for Cell Types Relevant in Clinical Research Recorded: Apr 24 2014 42 mins
    Deepa Saxena
    Expansion of stem cells and some primary cells requires either animal-origin components in culture media or coating of the culture vessels with human or animal-derived extracellular matrix (ECM) protein. Growing concerns about introducing human and animal-derived pathogens into the culture necessitate the need for animal- free (xeno-free and human-origin components-free) culture environment. Herein, we present defined and synthetic surfaces for cell culture. Corning® PureCoat™ ECM Mimetic cultureware collagen I peptide and fibronectin peptide support culture of cell types requiring collagen I or fibronectin protein -coated surfaces. Corning Synthemax® surface consists of a vitronectin peptide and supports undifferentiated expansion of human pluripotent stem cells (hPSC) and neural progenitor cells (NPCs). ECM Mimetic and Synthemax cultureware are pre-coated, synthetic, animal-free, and room temperature stable. Synthemax-SC is available for self-coating.

    We demonstrate expansion and functionality of several clinically relevant cell types including hPSCs, mesenchymal stem cells (MSCs), human keratinocytes (HKN), endothelial colony forming cells (ECFCs), NPCs and cell lines for biomanufacturing (CHO and Vero) on these surfaces. Corning Synthemax and ECM Mimetic cultureware are versatile surfaces compatible with multiple media for culture of various cell types providing a ready to use alternative to ECM protein coating where animal-free and defined conditions are desirable.
  • Corning TransportoCells™, A Novel “Thaw and Go” Cell-based Transporter Model Recorded: Mar 31 2014 54 mins
    Na Li
    SLC transporter-mediated drug–drug interactions (DDI) can significantly impact the pharmacokinetics and safety profiles of drugs. The regulatory agencies (FDA/EMA) recent guidance documents recommend testing six SLC transporters for potential DDI: OATP1B1, OATP1B3, OAT1, OAT3, OCT1 and OCT2. The 2013 drug transporter white paper published by the International Transporter Consortium (ITC) identified additional drug transporters relevant to drug development, including the Multidrug and Toxin Extrusion SLC transporters: MATE1 and MATE2-K. The webinar will introduce a novel cell-based SLC transporter model system - the recently launched “Corning™ TransportoCells™” - for studying regulatory agency recommended SLC transporters. The new system provides a convenient “thaw and go”, high performing mammalian cell model which supports regulatory agency recommendations for evaluating transporter mediated drug-drug interactions in vitro. In this webinar, we will provide an overview of the product validation and applications for the TransportoCells™ transporter model system. Validation data will also be presented for the newly available MATE1 and MATE2-K Corning™ TransportoCells™.
  • Materials and Systems for Optimizing 3D Cell Culture Environments Recorded: Feb 18 2014 64 mins
    Marshall Kosovsky, Ph.D.
    Three-dimensional (3D) cell culture environments provide structural and biochemical cues for cellular differentiation and functionality. For specialized cell types such as primary cells, a two-dimensional growth substrate may not be sufficient to support complex cellular behaviors such as cell polarity, morphology, signal transduction, and tissue-specific gene expression. This webinar will highlight applications that rely on 3D materials and systems, including Corning ® Matrigel® Matrix (reconstituted basement membrane), Collagen Type I, Corning PuraMatrix™ Peptide Hydrogel, and permeable supports (cell culture inserts). To demonstrate the effectiveness of these environments, the biological and functional properties of a variety of cell types will be discussed.
  • Development of Hydrogel-Based 3D Cancer Stem Cell & Tumorsphere Models for HTS Recorded: Dec 3 2013 48 mins
    Lesley A. Mathews Griner and Madhu Lal-Nag
    Cancer stem cells (CSCs), commonly referred to as tumor initiating cells (TICs), are thought to be the origin of replicating malignant cells that remain after the primary tumor is removed or eradicated. While the contribution of TICs to cancers of hematopoietic origin is well established, there is a growing body of evidence that supports the presence of TICs within solid tumors. This heterogeneity contributes to the intrinsic resistance of solid tumors to chemotherapeutics and eventually leads to therapeutic failure and patient death. Historically, screening for oncology directed compounds has been performed in 2-dimensional monolayer cultures which fail to replicate the complex architecture and microenvironment of tumors in vivo. To address this need using prototype manufactured 384-well round bottom clear black plates coated with hydrogel (commonly referred to as ultra-low attachment or ULA plates) we have developed a method to generate single cell type spheroids to accommodate their use in high throughput screening. These cultures were systematically compared to spheres grown in conventional plates and comparative imaging analyses were carried out to assess sphere formation. In addition, using the same plates, we have developed multicellular tumor spheroids (MCTS) composed of various cell types cultured in the same well. Differences in the cellular compositions of these spheroids in response to drugs can be quantified using high content imagers such as the WiScan from Idea-Bio and the High Throughput Flow Cytometer from IntelliCyt. These heterotypic, differentially labeled spheroids are a valuable asset in the generation of cell-based HTS assays capable of identifying molecules that selectively kill TICs and the cells comprising their microenvironment.
  • Corning® TransportoCells™: Overview of a Novel SLC Transporter Model Recorded: Nov 14 2013 49 mins
    Na Li
    During the past decade, the significant role of transporters in drug disposition and safety has been strengthened due to the increasing evidence of transporter mediated drug-drug interaction (DDI). Regulatory agencies recommend evaluating specific drug transporters of emerging importance in drug development. Appropriate in vitro characterization of drug interactions with these transporters facilitates decision-making and development of appropriate clinical plans in drug development. Corning Life Sciences have recently developed a novel SLC transporter model to help the pharmaceutical industry address regulatory guidance in a fast and easy manner. In this webinar, we will provide an overview of the product validation and applications for the recently launched TransportoCells™ cryopreserved SLC transporter model.
  • Overview of In Vitro ABC Transporter Models and Applications Recorded: Oct 29 2013 64 mins
    Na Li
    During the past decade, increasing clinical evidence of transporter mediated drug-drug interaction (DDI) has highlighted the significant role transporters play in drug disposition and safety. Regulatory authorities recommend evaluating specific drug transporters of emerging importance during drug development, including SLC and ABC transporters. ABC transporters are more challenging to study because they are usually expressed on the apical side of cell membrane (i.e., ATP and substrate binding sites facing inside of the cell), which can limit the direct access of compounds to the transporters in the absence of a relevant uptake transport mechanism. It is critical to utilize an appropriate in vitro system to evaluate drug interactions with these transporters in order to gain better insight into the mechanisms of transporter mediated drug disposition and elimination.

    Corning Life Sciences offers a broad spectrum of in vitro models to support drug transporter studies. This webinar will provide an overview of in vitro ABC transporter models, as well as the pros and cons of each model in deciphering drug ADME features.
  • Cell Culture Contamination Part 3 - Strategies for Reducing Contamination Recorded: Sep 24 2013 56 mins
    Hilary Sherman
    In the third part of our three lecture series on cell culture contamination, we will focus on everyday management of cell culture contamination. We will discuss how to reduce cell culture accidents by using good aseptic technique, the importance of freeze stocks, and how to appropriately monitor for contamination. We will also learn how to track down the source of contamination to prevent future accidents.
Training for Life Science Researchers
The Corning Scientific Seminar Series is a series of free, online technical presentations that provide novel tips, best practices and proven techniques to help advance your research. Delivered by scientists to scientists, these one-hour broadcasts offer useful information and tips for lab technicians and researchers.

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  • Title: Cell Culture Contamination Part 1
  • Live at: Jan 25 2011 4:35 pm
  • Presented by: Jeanne Phillips, Ph.D.
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