Analysis of metabolic activity in cells using extracellular flux rate measurements
First Claim
1. A method for analysis comprising the steps of:
- incubating animal cells under analysis in a medium disposed in at least one of a plurality of wells in a multiwell plate;
adding to the medium to bring into contact with the cells a substance potentially capable of altering cellular metabolism; and
measuring in medium in a well the rate of change in concentration of both an extracellular solute which is a component of cellular aerobic metabolism and an extracellular solute which is a component of cellular anaerobic metabolism.
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Accused Products
Abstract
Disclosed are methods for non-destructively measuring in vitro the effect on cellular metabolism of the addition to animal cells in culture of a soluble molecule potentially capable of perturbing the biological state of the cells, such as a drug or drug candidate, a toxin, a ligand known or suspected to bind to a cell surface receptor, a nutrient, a cytokine, a growth factor, a chemokine, a metabolism inhibitor or stimulator. Also disclosed are methods for measuring cell viability, vitality, or quality, e.g., in anticipation of the execution of an experiment on the cells. The measurements are done by observing alteration in the rates of consumption or production of extracellular solutes related to aerobic and anaerobic cellular metabolism, such as oxygen, protons, nutrients, carbon dioxide, lactate, or lactic acid. The methods are particularly useful in drug discovery efforts, such as cancer drug discovery and searches for modulators of cellular metabolism.
239 Citations
34 Claims
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1. A method for analysis comprising the steps of:
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incubating animal cells under analysis in a medium disposed in at least one of a plurality of wells in a multiwell plate;
adding to the medium to bring into contact with the cells a substance potentially capable of altering cellular metabolism; and
measuring in medium in a well the rate of change in concentration of both an extracellular solute which is a component of cellular aerobic metabolism and an extracellular solute which is a component of cellular anaerobic metabolism. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 30, 31)
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12. The method of claim 111 comprising the additional steps of:
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measuring, in the medium of a cell culture in a well separate from said cells under analysis and treated differently than said cells under analysis, either or both the rate of change in concentration of an extracellular solute which is a component of cellular aerobic metabolism and an extracellular solute which is a component of cellular anaerobic metabolism; and
comparing the measurements of the rate of change in said separate cell culture to said cells under analysis.
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23. A method for analysis of cell culture quality comprising the steps of:
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measuring in a cell medium the rate of change in concentration of both an extracellular solute which is a component of cellular aerobic metabolism and an extracellular solute which is a component of cellular anaerobic metabolism; and
comparing said measured rates of change to a standard informative of known cell culture respiration rates thereby to assess the respiratory capacity, metabolic rate, or relative pathway utilization of the culture as a measure of cell vitality and cell quality. - View Dependent Claims (24, 25, 26, 27, 28, 29)
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32. A method for analysis comprising the steps of:
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a) exposing an animal to a test substance;
b) removing cells from said animal;
c) incubating said cells in a medium disposed in at least one of a plurality of wells in a multiwell plate;
d) measuring in the medium in the well the rate of change in concentration of both an extracellular solute which is a component of cellular aerobic metabolism and an extracellular solute which is a component of cellular anaerobic metabolism. - View Dependent Claims (33)
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34. A method for analysis comprising the steps of:
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incubating animal cells under analysis in a medium disposed in at least one of a plurality of wells in a multiwell plate;
subjecting said cells to a genetic alteration or environmental stress potentially capable of altering cellular metabolism; and
measuring in medium in a well the rate of change in concentration of both an extracellular solute which is a component of cellular aerobic metabolism and an extracellular solute which is a component of cellular anaerobic metabolism.
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Specification