Microbioreactor for continuous cell culture
First Claim
1. A microbioreactor comprising comprising:
- at least one culture vessel having an interior volume of less than 1 ml;
a mechanism for actively mixing the contents of the culture vessel;
an inflow port to allow fresh culture medium to be continuously supplied to the culture vessel; and
an outflow port to allow culture medium to be continuously removed from the culture vessel at the same rate as fresh medium is supplied, such that a constant fluid volume and constant growth conditions are maintained within the culture vessel for a prolonged period of time after cells cultured in the culture vessel reach a steady state.
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Abstract
The present invention microscale bioreactors (microfermentors) and microscale bioreactor arrays for use in culturing cells. The microfermentors include a vessel for culturing cells and means for providing oxygen to the interior of the vessel at a concentration sufficient to support cell growth, e.g., growth of bacterial cells. Depending on the embodiment, the microfermentor vessel may have various interior volumes of less than approximately 1 ml. The microfermentors may include an aeration membrane and optionally a variety of sensing devices. Methods of using the microfermentors, e.g., to select optimum cell strains or bioprocess parameters are provided. The microbioreactors having a variety of different designs, some of which incorporate active fluid mixing and/or have the capability to operate in batch, fed-batch, or continuous mode. In certain embodiments the microreactors operate as microchemostats. Methods for culturing cells under chemostat conditions in a microbioreactor are also provided.
197 Citations
195 Claims
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1. A microbioreactor comprising comprising:
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at least one culture vessel having an interior volume of less than 1 ml;
a mechanism for actively mixing the contents of the culture vessel;
an inflow port to allow fresh culture medium to be continuously supplied to the culture vessel; and
an outflow port to allow culture medium to be continuously removed from the culture vessel at the same rate as fresh medium is supplied, such that a constant fluid volume and constant growth conditions are maintained within the culture vessel for a prolonged period of time after cells cultured in the culture vessel reach a steady state. - View Dependent Claims (138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 187, 188, 189, 190)
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2-137. -137. (canceled)
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175. A method of performing cell culture comprising:
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introducing at least one cell into a microbioreactor that comprises a culture vessel having an interior volume of less than 1 ml;
continuously flowing fresh culture medium into the vessel while continuously removing culture medium containing cells from the culture vessel at the same rate as that with which fresh medium enters the vessel so that a constant medium volume is maintained in the culture vessel;
actively mixing the contents of the culture vessel;
maintaining the cells for sufficient time to achieve a first steady state. - View Dependent Claims (176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186)
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191. A method of modifying a polymeric surface other than a PDMS surface so as to confer resistance to adherence of cells, proteins, or both, comprising steps of:
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assembling an amine-terminated self assembled monolayer on the surface; and
contacting the surface with a PEG-containing polymer under conditions in which the PEG-containing polymer contains sufficient negative charges to cause adsorption to the self-assembled monolayer. - View Dependent Claims (192, 193, 194, 195)
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Specification