In the seahorse apparatus (Agilent) small changes in metabolic parameters of living cells seeded in a 96 mw plate are measured in real time. The apparatus, through two specific probes, allows the measurement of the oxygen consumption rate (OCR) and the acidification rate of the culture medium (ECAR), as respiration and glycolytic rate, respectively. These measurements are associated with the use of specific drugs that specifically target the key proteins of the main metabolic processes allowing the investigation of the critical metabolic functions of the selected cells in physiological conditions or after specific treatments. For each different project it is possible to select a specific sequence of drugs that allow to clarify in the best possible way the functionality of the selected metabolic pathways. Measurements are possible both under normoxic or hypoxic conditions (up to 3% oxygen).
- Cells must be checked for mycoplasma contamination before the shipping.
- For each cell type preliminary experiments are recommended relating to the number of cells to be seeded and the sensitivity of the same to the different drugs that will allow the reading to be set correctly.
What other specific considerations are relevant?
The growth conditions of each cell line being tested should be discussed with the cell culture managers. It may be required to send culture media or specific components (antibiotics, growth factors etc.) in case of particular cellular needs.
Partners offering this technique
MOSBRI reference partner site for this technique: