Using Seahorse Machine to Measure OCR and ECAR in Cancer Cells. | Semantic Scholar (2024)

Step-by-step procedures to isolate, culture, plate and run a seahorse assay for measuring cellular metabolism, and an example of oxygen consumption and extracellular acidification rate traces obtained from MC3T3E1-C4 cells using the XFe96 seahorses analyzer are provided.

A simple plate-based approach for real-time measurements of acid production and oxygen depletion under typical culture conditions that enable metabolic monitoring for extended periods of time and can deliver a robust appraisal of metabolism in cell lines, with applications in drug screening and in quantitative studies of metabolic regulation.

A platform that couples the metabolic flux assay with high-content fluorescence imaging to simultaneously enhance normalization of respiration data with cell number; analyze cell cycle progression; quantify mitochondrial content, fragmentation state, membrane potential, and mitochondrial reactive oxygen species is described.

Methods to evaluate the structural and functional parameters of mitochondrial health in the context of tumors and cancer cells are discussed, with a particular emphasis on gynecologic malignancies.

A platform that couples the metabolic flux assay with high-content fluorescence imaging to simultaneously provide means for normalization of respiration data with cell number; analyze cell cycle distribution; and quantify mitochondrial content, fragmentation state, membrane potential, and mitochondrial reactive oxygen species is described.

A platform that couples the metabolic flux assay with high-content fluorescence imaging to simultaneously provide means for normalization of respiration data with cell number; analyze cell cycle distribution; and quantify mitochondrial content, fragmentation state, membrane potential, and mitochondrial reactive oxygen species is described.

A simplified model of the metabolism which focuses on the interrelation of the three main energetic metabolites that are oxygen, glucose and lactate with the aim to better understand the dynamic of the core system of the glycolysis-OXPHOS relationship is proposed.

It is shown that both O2 levels and media composition significantly affect mitochondrial abundance and network structure, concomitantly with changes in cellular bioenergetics.

Bone powder, regardless of chicken treatment, contains and releases factors/chemicals responsible for the observed effects on energy metabolism, and Quantitative differential effects appear to depend on biochemical alterations in the bone matrix caused by antibiotics rather than antibiotics themselves.

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This unit contains several protocols to determine the energy utilization of T cells in real‐time using a Seahorse Extracellular Flux Analyzer, which allows for the analysis of immediate metabolic changes after T cell receptor stimulation.

Protocols for analyzing energy metabolism in hPSCs and their early differentiated progenies that are generally applicable to mature cell types as well are provided and Companion methods are provided to aid researchers in developing more sophisticated experimental regimens for extended analyses of cellular bioenergetics.

It is proposed that energy metabolism may be an alternative therapeutic target for both hypoxic (glycolytic) and oxidative tumors.

Using an innovative system, the total ATP turnover of the MCF-7 breast cancer cell line is measured, the contributions to this turnover by oxidative and glycolytic ATP production and the contributes to the oxidative component by glucose, lactate, glutamine, palmitate and oleate are measured.

It is concluded that, in the AS-30D hepatoma cell line, glucose is the preferred energy source, with the larger portion of ATP production being supplied by glycolytic reactions.

It is reported that the major function of glucose metabolism for Kras-induced anchorage-independent growth, a hallmark of transformed cells, is to support the pentose phosphate pathway.

Cancer cells then reprogramme adjacent stromal cells to optimize the cancer cell environment and activate out-of-context programmes that are important in development, stress response, wound healing and nutritional status.

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