How to Keep Calm and Monitor Cell Stress, Death, and Behavior

Tuesday, November 21, 2017 - 14:57

Measuring and monitoring cell health, stress, and behavior is a necessary part of using 2D and 3D cell culture models. This can be done qualitatively through microscopic assessment of cell structure, morphology, and confluency. Quantitative assessment of cell stress, death, and behavior is often needed when using 2D or 3D cell cultures for drug discovery, drug screening, and disease modeling. With the acquisition of Trevigen, Bio-Techne now has an extensive portfolio and unmatched expertise in assays and reagents for measuring cellular proliferation, viability, stress, DNA damage, and apoptosis. This blog showcases our expertise by highlighting the various resources and tools available through Bio-Techne.

DNA Damage

As part of the Trevigen acquisition, Bio-Techne becomes an expert in DNA damage, an area where Trevigen has developed and optimized quantitative high throughput assays for both screening and detection. Specifically, we now offer a full CometAssay^® portfolio of reagents, equipment, and software to quantify DNA damage in single cells. Trevigen improved the reproducibility of the traditional CometAssay^® with a complete and standardized system for consistent results. The new CometChip^® platform increases throughput 10,000% by providing high density data using a cellular array. In addition, their gamma-H2AX Kit enables the indirect measurement and confirmation of DNA double-strand breaks in an ELISA format. Trevigen also brings to Bio-Techne important drug discovery screening tools for inhibitors to PARP-mediated DNA repair, potential targets in cancer therapy. While these DNA damage tools are not entirely new to Bio-Techne - R&D Systems has sold many of them for the greater part of a decade - we now offer Trevigen’s extended portfolio to our constituents.

Apoptosis and Cell Viability

Our portfolio of tools to monitor apoptosis and viability spans all Bio-Techne brands. For example, Novus Biologicals has an Apoptosis Handbook that can be used as a mechanistic and scientific reference to find antibodies and assays. In particular, Novus Biologicals offers pSIVA, a robust reagent for the real-time measurement of phosphatidylserine (PS) externalization, a cell membrane event observed during apoptosis. R&D Systems is also a reliable scientific resource of cell proliferation and viability assays, including Calcein AM assays and Annexin V kits. This selection is expanded through the Trevigen acquisition, bringing to the Bio-Techne portfolio a broader offering of kits for Annexin V, TUNEL Assays, and MTT and XTT cell proliferation/viability kits. Last, through Tocris Bioscience, Bio-Techne offers an extended list of compounds to monitor and modulate pathways involved in apoptosis.

Oxidative Damage

The generation of reactive oxygen species (ROS) and reactive nitrogen species (RNS) occurs in many tissues following injury, infection, and inflammation. The impact of oxidative damage due to ROS and RNS is recognized as key contributor to neurodegenerative disease, tissue injury, cancer, and the aging process. Optimizing and controlling oxidative stress is also an important phenomenon to consider for maintaining health in routine cell culture conditions. Tocris Bioscience offers a number of fluorescent probes to assist research in monitoring in vitro and in vivo ROS and RNS generation. These probes are complimented by the more extensive selection of tools to monitor oxidative damage offered by Trevigen, including antibodies and assays to measure 8-hydroxy 2-deoxyguanisine (8-oxo-dG), superoxide dismutase (SOD), and glutathione activity. Specifically, the 8-oxo-dG ELISA Kit is a species independent competitive ELISA validated for many samples types including DNA, plasma, urine, and saliva. An 8-oxo-dG antibody is also available for independent ELISA and immunocytochemistry experiments. The Superoxide Dismutase Assay Kits allow for quantitation of SOD in mammalian cell and tissue extracts in a high-throughput 96-well format. Last, the Glutathione Assay Kits quantify oxidized and reduced glutathione levels, glutathione reductase activity, and glutathione peroxidase activity in 96-well formats.

Cell Proliferation and Migration/Invasion Assays

Cultrex^® assays for cell proliferation, migration, and invasion were referenced in our blog post discussing the merits of Cultrex^® Basement Membrane Extracts (BME). These assays utilize BME and other extracellular matrix proteins (ECM) providing the research community with standardized and quantifiable assays to monitor cell behavior. These assays have been incorporated into the workflow of R&D Systems and are featured in a peer-reviewed publication in the Journal of Visualized Experiments, that discusses a novel media for the induction of epithelial-to-mesenchymal transition (EMT). To verify EMT induction, R&D Systems scientists employed the Cultrex^® cell migration assay to quantify motility of mesenchymal cells generated using the EMT Induction Media. Watch the video publication here.

Keep Calm and Monitor Cell Stress, Death, and Behavior

As detailed in the extensive breakdown of cell stress, death, and behavior assays above, Bio-Techne is making cell monitoring more accessible to the research community. When looking for good solutions, just remember to keep calm and go to Bio-Techne to find the right cell stress, death, and behavior assay.

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Stem Cells