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US 7,247,470 B2Granted
Method and apparatus for measuring nitric oxide production and oxygen consumption in cultures of adherent cells
AssigneeThe Trustees of the University of Pennsylvania
InventorsJames E. Baumgardner, Cynthia M. Otto
Life of this patent
Estimating the term…
Abstract
An apparatus and method for measuring nitric oxide production and oxygen consumption in cultures of adherent cells continuously and without destroying the cells. The method involves flowing growth media through a tube having adherent cells are adhered to the inner surface thereof and then contacting the growth media with an NO or O2 sensor to detect the concentration of NO or O2 in the growth media.
Claims26
A nitric oxide (NO) measuring apparatus comprising:
- a first gas equilibrator having an inlet and an outlet, the first gas equilibrator configured to maintain a first concentration of a first gas within a first fluid, when the first fluid flows through the first gas equilibrator and exits the first gas equilibrator as a first effluent;
- a second gas equilibrator having an inlet and an outlet, the second gas equilibrator configured to maintain a second concentration of a second gas within a second fluid, when the second fluid flows through the second gas equilibrator and exits the second gas equilibrator as a second effluent;
- a tubular member configured to accommodate passage of a fluid therethrough, the tubular member having an inner surface suitable for having living cells adhered thereto, said tubular member having an inlet and an outlet, wherein the inlet of the tubular member is in fluid communication with the outlet of the first gas equilibrator, when the apparatus is in a first configuration;
- a first NO sensor in fluid communication with the first gas equilibrator via the tubular member, the first NO sensor configured to measure an NO concentration of the first effluent from the first gas equilibrator after the first effluent has flowed through the tubular member, when the apparatus is in said first configuration;
- a first oxygen (O2) sensor in fluid communication with the second gas equilibrator, the first oxygen sensor configured to measure an O2 concentration of the second effluent from the second gas equilibrator without the second effluent having flowed through the tubular member, when the apparatus is in said first configuration;
- and
- one or more valves directing the first effluent to the inlet of the tubular member and thus to the first NO sensor, and directing the second effluent to the first O2 sensor, when the apparatus is in said first configuration.
4 more elements
A nitric oxide (NO) measuring apparatus comprising:
- a first gas equilibrator having an inlet and an outlet, the first gas equilibrator configured to maintain a first concentration of a first gas within a first fluid, when the first fluid flows through the first gas equilibrator and exits the first gas equilibrator as a first effluent;
- a second gas equilibrator having an inlet and an outlet, the second gas equilibrator configured to maintain a second concentration of a second gas within a second fluid, when the second fluid flows through the second gas equilibrator and exits the second gas equilibrator as a second effluent;
- a tubular member configured to accommodate passage of a fluid therethrough, the tubular member having an inner surface suitable for having living cells adhered thereto, said tubular member having an inlet and an outlet, wherein the inlet of the tubular member is in fluid communication with the outlet of the first gas equilibrator, when the apparatus is in a first configuration;
- a first NO sensor in fluid communication with the first gas equilibrator via the tubular member, the first NO sensor configured to measure an NO concentration of the first effluent from the first gas equilibrator after the first effluent has flowed through the tubular member, when the apparatus is in said first configuration;
- a first oxygen (O2) sensor in fluid communication with the second gas equilibrator, the first oxygen sensor configured to measure an O2 concentration of the second effluent from the second gas equilibrator without the second effluent having flowed through the tubular member, when the apparatus is in said first configuration;
- and
- a first valve in fluid communication with the first and second gas equilibrators, the first valve configured to:
- receive the first effluent from the first gas equilibrator;
- receive the second effluent from the second gas equilibrator;
- and
- supply one of the first and second effluents to the inlet of the tubular member while directing the other of the first and second effluents to a bypass around the tubular member.
8 more elements
A nitric oxide (NO) measuring apparatus comprising:
- a first gas equilibrator having an inlet and an outlet, the first gas equilibrator configured to maintain a first concentration of a first gas within a first fluid, when the first fluid flows through the first gas equilibrator and exits the first gas equilibrator as a first effluent;
- a second gas equilibrator having an inlet and an outlet, the second gas equilibrator configured to maintain a second concentration of a second gas within a second fluid, when the second fluid flows through the second gas equilibrator and exits the second gas equilibrator as a second effluent;
- a third gas equilibrator having an inlet and an outlet, the third gas equilibrator configured to maintain a third concentration of a third gas within a third fluid, when said third fluid flows through the third gas equilibrator, wherein an effluent of the third gas equilibrator merges with the second effluent to thereby form a blended effluent;
- a tubular member configured to accommodate passage of a fluid therethrough, the tubular member having an inner surface suitable for having living cells adhered thereto, said tubular member having an inlet and an outlet, wherein the inlet of the tubular member is in fluid communication with the outlet of the first gas equilibrator, when the apparatus is in a first configuration;
- a first NO sensor in fluid communication with the first gas equilibrator via the tubular member, the first NO sensor configured to measure an NO concentration of the first effluent from the first gas equilibrator after the first effluent has flowed through the tubular member, when the apparatus is in said first configuration;
- a first oxygen (O2) sensor configured to measure an O2 concentration of the blended effluent without the blended effluent having flowed through the tubular member, when the apparatus is in said first configuration;
- and
- one or more valves directing the first effluent to the inlet of the tubular member and thus to the first NO sensor, and directing the blended effluent to the first O2 sensor, when the apparatus is in said first configuration.
5 more elements
A gas measuring apparatus comprising:
- a first gas equilibrator configured to maintain a first concentration of a gas within a first fluid;
- a second gas equilibrator configured to maintain a second concentration of a gas within a second fluid;
- a switching valve in fluid communication with the first gas equilibrator and the second gas equilibrator;
- a tubular member having an inlet in fluid communication with an outlet of the switching valve, wherein the first gas and the second gas are separately and selectively fed through the switching valve to the tubular member;
- a first gas sensor in fluid communication with an outlet of the tubular member, the first gas sensor being one of an NO or an O2 sensor;
- and
- a second gas sensor in fluid communication with the switching valve through a bypass around the tubular member, the second gas sensor being the other of an NO or an O2 sensor.
4 more elements
A nitric oxide (NO) measuring apparatus comprising:
- a first gas equilibrator having an inlet and an outlet, the first gas equilibrator configured to maintain a first concentration of a first gas within a first fluid, when the first fluid flows through the first gas equilibrator and exits the first gas equilibrator as a first effluent;
- a second gas equilibrator having an inlet and an outlet, the second gas equilibrator configured to maintain a second concentration of a second gas within a second fluid, when the second fluid flows through the second gas equilibrator and exits the second gas equilibrator as a second effluent;
- a tubular member configured to accommodate passage of a fluid therethrough, the tubular member having an inner surface suitable for having living cells adhered thereto, said tubular member having an inlet and an outlet, wherein the inlet of the tubular member is selectively connectable to the outlet of one of the first and second gas equilibrators;
- a first NO sensor selectively connectable to the outlet of the tubular member and configured to measure an NO concentration of one of the first and second effluents;
- and
- a first oxygen (O2 ) sensor selectively connectable to the outlet of the tubular member and configured to measure an O2 concentration of the other of the first and second effluents;
- wherein:
- one or more valves selectively connect:
- the inlet of the tubular member to the outlet of one of the first and second gas equilibrators, the first NO sensor to the outlet of the tubular member and the first O2 sensor to the outlet of the tubular member, such that the apparatus is capable of selectively occupying any of a plurality of configurations including:
- a first configuration in which the first effluent passes through the tubular member and the NO sensor measures an NO concentration of said first effluent, while the second effluent bypasses the tubular member and the O2 sensor measures an O2 concentration of the second effluent;
- a second configuration in which the first effluent passes through the tubular member and the O2 sensor measures an O2 concentration of said first effluent, while the second effluent bypasses the tubular member and an NO sensor measures an NO concentration of the second effluent;
- a third configuration in which the second effluent passes through the tubular member and the NO sensor measures an NO concentration of said second effluent, while the first effluent bypasses the tubular member and the O2 sensor measures an O2 concentration of the first effluent;
- and
- a fourth configuration in which the second effluent passes through the tubular member and the O2 sensor measures an O2 concentration of said second effluent, while the first effluent bypasses the tubular member and an NO sensor measures an NO concentration of the first effluent.
11 more elements
A nitric oxide (NO) measuring apparatus comprising:
- a first gas equilibrator having an inlet and an outlet, the first gas equilibrator configured to maintain a first concentration of a first gas within a first fluid, when the first fluid flows through the first gas equilibrator and exits the first gas equilibrator as a first effluent;
- a second gas equilibrator having an inlet and an outlet, the second gas equilibrator configured to maintain a second concentration of a second gas within a second fluid, when the second fluid flows through the second gas equilibrator and exits the second gas equilibrator as a second effluent;
- a third gas equilibrator having an inlet and an outlet, the third gas equilibrator configured to maintain a third concentration of a third gas within a third fluid, when said third fluid flows through the third gas equilibrator, wherein an effluent of the third gas equilibrator merges with the second effluent to thereby form a blended effluent;
- a tubular member configured to accommodate passage of a fluid therethrough, the tubular member having an inner surface suitable for having living cells adhered thereto, said tubular member having an inlet and an outlet, wherein the inlet of the tubular member is selectively connectable to receive either the first effluent or the blended effluent;
- a first NO sensor selectively connectable to the outlet of the tubular member and configured to measure an NO concentration of one of the first and blended effluents;
- and
- a first oxygen (O2) sensor selectively connectable to the outlet of the tubular member and configured to measure an O2 concentration of the other of the first and blended effluents;
- wherein:
- one or more valves selectively connect:
- the inlet of the tubular member to receive either the first effluent or the blended effluent, the first NO sensor to the outlet of the tubular member and the first O2 sensor to the outlet of the tubular member such that the apparatus is capable of selectively occupying any of a plurality of configurations including:
- a first configuration in which the first effluent passes through the tubular member and the NO sensor measures an NO concentration of said first effluent, while the blended effluent bypasses the tubular member and the O2 sensor measures an O2 concentration of the blended effluent;
- a second configuration in which the first effluent passes through the tubular member and the O2 sensor measures an O2 concentration of said first effluent, while the blended effluent bypasses the tubular member and an NO sensor measures an NO concentration of the blended effluent;
- a third configuration in which the blended effluent passes through the tubular member and the NO sensor measures an NO concentration of said blended effluent, while the first effluent bypasses the tubular member and the O2 sensor measures an O2 concentration of the first effluent;
- and
- a fourth configuration in which the blended effluent passes through the tubular member and the O2 sensor measures an O2 concentration of said blended effluent, while the first effluent bypasses the tubular member and an NO sensor measures an NO concentration of the first effluent.
12 more elements
Dependencies are inferred from the claim text.