Method and device for cell culture in open continuous mode
AssigneeEco Solution
InventorBernard KUDLA
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Abstract
The present invention relates to the use of cell cultures in the open continuous mode, to a method for selecting static cell variants or cell variants which proliferate in suspension, to a culture substrate and to a device suitable for implementing this method.
Claims47
Method for continuous cell culture allowing the selection of cell variants that are static or proliferating in suspension, characterized in that it comprises the following stages:
- a) a liquid culture medium contained in a first culture vessel kept open in a closed container is seeded using one or more live cells,
- b) said cells, in said culture medium, are brought to a specific growth stage, corresponding to a given cell density or to a physicochemical parameter measured in the culture medium,
- c) the cell density in the culture medium, or the value of said physicochemical parameter, reached in stage b), is kept substantially constant by supplying fresh culture medium or a diluent in said culture vessel,
- d) a portion of the culture medium of the culture obtained in c) containing the cells in suspension is removed by pipetting so as to maintain the volume of the culture,
- e) a fraction of the culture medium obtained in d) is transferred into a second culture vessel,
- f) said first culture vessel with the remaining culture fraction that it contains is withdrawn;
- g) after several generations of culture in the second vessel, the cells proliferating in suspension and/or the static cell variants are selected.
Method for continuous cell culture allowing the selection of cell variants proliferating in suspension, characterized in that it comprises the following stages:
- a) a liquid culture medium contained in a first culture vessel kept open in a closed container is seeded using one or more live cells,
- b) the cells, in said culture medium, are brought to a specific growth stage, corresponding to a given cell density or to a physicochemical parameter measured in the culture medium,
- c) the cell density of the culture, or the value of said physicochemical parameter, reached in stage b), is kept substantially constant by supplying fresh culture medium or at least one diluent in said culture vessel,
- d) a portion of the culture medium containing the cells in suspension is removed by pipetting so as to maintain the volume of the culture,
- e) a fraction of the culture obtained in d) in which the cells are in suspension, is transferred to a second culture vessel replacing the first;
- f) said first culture vessel with the remaining culture fraction that it contains is withdrawn;
- g) after several generations of culture in the second vessel, the cells proliferating in suspension are selected.
Method for continuous cell culture allowing the selection of static cell variants, characterized in that it comprises the following stages:
- a) a liquid culture medium contained in a first culture vessel is seeded using one or more live cells, in which at least one solid surface, preferably a plate, is placed, said vessel being kept open in a closed container,
- b) the cells, in the seeded culture medium, are brought to a specific growth stage, corresponding to a given cell density or to a physicochemical parameter measured in the culture medium,
- c) the cell density of the culture, or the value of said physicochemical parameter, reached in stage b), is kept substantially constant by supplying fresh culture medium or at least one diluent in said culture vessel,
- d) a portion of the culture medium obtained in c) containing the cells in suspension is removed by pipetting so as to maintain the volume of the culture,
- e) said solid surface on which a fraction of the culture obtained in d) is deposited, is transferred to a second culture vessel replacing the first,
- f) said first culture vessel with the remaining fraction of culture that it contains is withdrawn,
- g) after several generations of culture, the static cell variants adhering to said solid surface are selected.
Method according to the previous claim, characterized in that the sterile gas stream from the top to the bottom is maintained by creating a partial vacuum at the periphery of the culture vessel.
Culture support (18) making it possible to carry out a cell culture in open continuous mode, characterized in that it comprises:
- at least one culture vessel (1) open at the top (6) suitable for containing a liquid culture medium;
- at least one containment tank (2) open towards the top, in which said culture vessel is housed (1);
- a space (7) between said culture vessel (1) and said containment tank (2), suitable for allowing a gas stream to circulate at the periphery of the opening of the vessel, from the top to the bottom, between the culture vessel (1) and the inner walls of the containment tank (2);
- and
- a means for extracting (8) said gas stream, situated in the lower part of the containment tank (2).
Dependencies are inferred from the claim text.