Method for gas enrichment and simultaneously for displacement of a fluid, and system for controlling the cell environment on a corresponding multi-well cell culture plate
A method displacing a fluid and simultaneously gas enriching a liquid cell culture medium with a gas. The method includes injecting a controlled volume of a gas or gas mixture into a one chamber by using a gas flow controller, the injection taking place through a gas inlet into a volume of liquid. This injection produces bubbling and agitation of the volume of liquid; a build-up of gas or gas mixture due to buoyancy in a hermetic space formed by the volume of liquid and the chamber, and a pressure increase in the chamber until a sufficient controlled pressure is reached of less than or equal to 10 bar. This increase displaces the volume of liquid by a fluid outlet connecting the volume of liquid to the exterior of the chamber. Also provided are a device implementing the method and a cell culture system in a multi-well culture plate.
1. A method for displacement of a fluid and simultaneously for gas enrichment of a liquid cell culture medium, wherein the method comprises:
injecting a controlled volume of a gas or gas mixture into at least one pressure chamber by at least one gas flow controller, the injecting being performed through at least one gas inlet into a volume of the liquid cell culture medium, the injecting producing bubbling and agitation of said volume of liquid cell culture medium;
building up said gas or gas mixture due to buoyancy in a hermetic space formed by said volume of liquid cell culture medium and said at least one pressure chamber, and
increasing pressure in said at least one pressure chamber until a sufficient controlled pressure is reached of less than or equal to 10 bar, said increasing pressure producing a displacement of said volume of liquid cell culture medium through at least one fluid outlet connecting said volume of liquid cell culture medium to an exterior of said at least one pressure chamber.
2. The method according to claim 1 , wherein said at least one injected gas is chosen from among O 2 , CO 2 , N 2 or their mixtures.
3. The method according to claim 1 , wherein said displacement of said volume of liquid cell culture medium by said at least one fluid outlet connecting said volume of liquid cell culture medium to the exterior of said at least one pressure chamber is towards a microfluidic device.
4. The method according to claim 1 , wherein said volume of liquid cell culture medium is successively displaced through several pressure chambers mounted in series or in parallel with each other.
5. The method according to claim 1 , wherein said controlled volume of a gas or gas mixture is injected into an isolated individual reservoir forming said at least one pressure chamber.
6. The method according to claim 3 , wherein the method further comprises:
adapting said microfluidic device to a multi-well cell culture plate, so as to:
form said at least one pressure chamber from at least one well of said multi-well cell culture plate and at least one cavity comprised in a manifold of said microfluidic device,
form at least one complete microfluidic circuit extending from at least one connection orifice or at least one fluidic connection duct to said manifold, by combining at least one microfluidic channel formed in at least one integrated microfluidic plate and at least one nozzle or at least one orifice or at least one duct of said manifold, said at least one nozzle or at least one orifice or at least one duct extending said at least one microfluidic channel, said volume of liquid cell culture medium being displaced through said microfluidic circuit.