Cellular Metabolism

Cayman’s cellular metabolism screening services can be used to provide a detailed view of cellular metabolic function via preclinical analysis of mitochondrial toxicity, function, and biogenesis, as well as glycolysis, metabolic substrate utilization, oxidative stress, and more. Our assays can be conducted using a variety of systems ranging from primary cells to isolated mitochondria and can be used to demonstrate improvements in metabolic function or to de-risk potential toxins. Our scientists are also experts in the design of biochemically relevant experimental models to investigate changes in cellular metabolism in the context of cancer, metabolic syndrome, NASH/NAFLD, and other bioenergetically challenging disorders. We are eager to assist you in the design and execution of cellular metabolism experiments and the development of model systems to meet your project’s needs.

Applications

• Mitochondrial toxicity assessment and preclinical compound de-risking
• Measure improvements in mitochondrial biogenesis and function in various disease models
• Measure cellular oxidative stress or mitochondrial reactive oxygen species (ROS) generation
• Immunometabolism studies (metabolism and energetics of immune cells)

Full-Service Capabilities

All studies are compatible with both primary cells and established cell lines. Our primary cell competencies include, but are not limited to, cardiomyocytes, inducible pluripotent stem cells (iPSCs), PBMCs, macrophages, neutrophils, pre- and mature adipocytes, and fibroblasts. We can also isolate mitochondria from cell lines and tissues, creating a simplified and biochemically relevant experimental model system.

Mitochondrial Toxicity Screening

Determining the effects of preclinical compounds on mitochondrial toxicity is a valuable tool in de-risking potential therapeutics.

Cayman offers comprehensive mitochondrial toxicity screening services using state-of-the-art equipment and assays. Mitochondrial toxicity is initially assessed by measuring mitochondrial oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) using the industry standard Agilent Seahorse XF Pro Analyzer. For compounds found to inhibit oxidative phosphorylation, the mechanism of toxicity is further refined by focusing on the individual components of the electron transport chain (ETC) using Cayman’s line ofMitoCheck^® assays. MitoCheck^® assays are designed to measure the activity of complexes I-IV of the ETC and the mitochondrial ATP synthase (also known as complex V) and are tailored specifically to each complex using distinct sets of substrates and inhibitors.

Following assessment of mitochondrial toxicity, additional functional assays can be used to further elucidate the effects of test compounds on mitochondrial and whole-cell function.

Metabolic and Functional Analysis of Mitochondria or Whole Cells

Cayman’s scientists can examine the effects of your compound(s) of interest on a variety of functional and energetic outcomes at the mitochondrial and whole-cell levels. Our methods can be applied to isolated mitochondria, cellular models of disease, and a variety of cell types, including immune cells.

Mitochondrial Function and Biogenesis

For several diseases, therapeutic avenues include upregulation of mitochondrial biogenesis and improvement of mitochondrial function. Mitochondrial function and biogenesis can be assessedvia:

• Mitochondrial stress test (OCR/ECAR)
• High-content imaging of mitochondrial morphology and/or content
• Fatty acid oxidation analysis (β-oxidation of various fatty acid substrates)
• Mitochondrial inner membrane permeability assays
• Measurement of mitochondrial membrane potential

Whole-Cell and Multi-Level Analysis

To determine the effects of experimental compounds or systems on whole-cell health, function, and bioenergetics, our experts can employ a variety of methods including:

• Cell viability and probe-free growth rate assessment
• Glycolysis/glycolytic flux analysis
• ATP quantification
• Evaluation of oxidative stress using the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG) as a readout
• Substrate utilization studies (e.g., glutamate, pyruvate, or long-chain fatty acid uptake/transport)
• Lipid droplet content and trafficking
• Lysosomal acidification and colocalization studies

Several phenotypes can be measured at either the mitochondrial or whole-cell level, such as:

• ROS generation
• Changes in additional oxidative stress markers
• Altered gene expression

Instrumentation

Pricing

For more information about our cellular metabolism services and project-specific pricing, please submit the “Contact Us” form.

View the Posters

• Screening a Kinase Inhibitor Library for Mitochondrial Dysfunction
• Response of Metabolic Phenotype to Chemotherapy Agents in a Simulated Tumor Microenvironment
• Cell-Based Metabolic Screening under Physiologically Relevant Oxygen Concentrations
• Characterization of Metabolic Changes Induced by Anticancer Drugs in MCF-7 and MDA-MB-231 Cell Lines

Read the Application Notes

• Mitochondrial Fuel Flexibility Assessment in a Tumor-Relevant Model
• O₂ Concentration of a Simulated Tumor Environment Affects Metabolic Response to Chemotherapeutics
• Characterizing a Better Uncoupler
• Screening for Drug Induced Mitochondrial Dysfunction

Read the Articles

• Manipulating Mitochondrial Fuel Pathways
• Measure Mitochondrial H₂O₂ Directly In Vivo or in Cells

View the Webinar

• Tools for Characterizing Metabolic Change in Cell-Based Systems

Cayman Collaborations & Co-Authored Publications

• Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS): Where will the drugs come from?