WO2019206207A1 - Ensemble de ventilation active, bioréacteur de type ventilation active et dispositif de culture de cellules - Google Patents

Ensemble de ventilation active, bioréacteur de type ventilation active et dispositif de culture de cellules Download PDF

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Publication number
WO2019206207A1
WO2019206207A1 PCT/CN2019/084203 CN2019084203W WO2019206207A1 WO 2019206207 A1 WO2019206207 A1 WO 2019206207A1 CN 2019084203 W CN2019084203 W CN 2019084203W WO 2019206207 A1 WO2019206207 A1 WO 2019206207A1
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Prior art keywords
gas
bioreactor
active
cell culture
container
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PCT/CN2019/084203
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English (en)
Chinese (zh)
Inventor
韩志强
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Shanghai Jiubo Biotechnology Co Ltd
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Shanghai Jiubo Biotechnology Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/24—Recirculation of gas
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/34—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of gas

Definitions

  • the present invention relates to a preparation and processing apparatus for biological materials, and more particularly to a living ventilator for a bioreactor and a cell culture device, and a cell culture device and a bioreactor having the bioreactor and the active ventilating assembly for the cell culture device.
  • the cell culture device is a container for culturing cells, such as a cell culture flask, a multi-layer plate culture flask, a multi-layer cell culture device, a cell factory, and a cell culture bag (for example, made of a gas permeable but water-impermeable film material).
  • the cell culture device can be placed in an incubator, and the gas in the incubator is previously configured to be suitable for cell culture.
  • the gas in the cell culture vessel can be exchanged with the gas in the incubator.
  • the level of liquid in the cell culture device is typically low (eg, 1-2 ml/cm 2 ). Therefore, although the gas exchange mode is a dispersion exchange, the transfer of oxygen and nutrients from the incubator required for cell culture can be achieved without agitation of the liquid in the cell culture device.
  • a bioreactor is a container in which biological cells and their biologically active substances undergo a chemical reaction.
  • the bioreactor can be placed in an incubator, the gas in the incubator being pre-configured to be suitable for the chemical reaction. It is often desirable to drive the flow of the reaction liquid in the bioreactor to effect the transfer of oxygen and nutrients required for the chemical reaction from the incubator.
  • the bioreactor can be a suspension culture bioreactor such as a stirred tank bioreactor, a soft bag bioreactor (eg, made of a gas impermeable, watertight barrier film material) and an airlift bioreactor.
  • a biological incubator is a laboratory device in which biological culture (e.g., cell culture) or biological reaction is performed.
  • the biological incubator includes a constant temperature incubator, a carbon dioxide incubator, and a three-gas incubator.
  • the constant temperature incubator uses a clean ambient air for cell culture.
  • the carbon dioxide incubator uses a clean ambient air and a certain proportion of carbon dioxide gas for cell culture.
  • the three-gas incubator uses a certain proportion of clean gas mixture of O 2 , CO 2 and N 2 for cell culture.
  • a certain partial pressure of carbon dioxide is required to maintain a stable pH to satisfy the activity of the cells.
  • the carbon dioxide content of the gas in the incubator needs to be maintained between 2 and 10% (e.g., 5%) to maintain the concentration of carbon dioxide to be dissolved in the culture solution.
  • the concentration of carbon dioxide in the air is very low. If the cells are not cultured in a carbon dioxide incubator, the HCO 3 - in the culture solution will be depleted, which will affect the normal growth of the cells. Therefore, the cultivation of most animal cells requires a carbon dioxide incubator or a three-gas incubator. Therefore, achieving accurate gas ratio formulation is important to meet cell growth. Thanks to the mature CO 2 and O 2 sensor technology, the biological incubator is generally equipped with two sensors, CO 2 and O 2 , and the concentration of nitrogen is 100%-CO 2 concentration - O 2 concentration.
  • a CO 2 sensor and a controller are disposed in the carbon dioxide incubator to detect the CO 2 concentration in the tank, and the detection result is transmitted to a control device such as a controller and a solenoid valve.
  • the solenoid valve is automatically opened, so that the CO 2 enters the tank from an external CO 2 gas source (for example, a cylinder in which CO 2 is stored).
  • the solenoid valve closes.
  • the CO 2 gas at the bottom of the tank can be thoroughly mixed with the air by a gas mixing pump, and then injected into the tank again, thereby avoiding stratification or unevenness of CO 2 .
  • Carbon dioxide incubators do not provide a gaseous environment for culturing cells that require high oxygen (higher than 22%) or hypoxic (less than 20%). Therefore, if you need to further control the concentration of oxygen in the tank, you need to use a three-gas incubator.
  • the three-gas incubator adds an oxygen and nitrogen inlet to the carbon dioxide incubator and is equipped with an oxygen concentration sensor.
  • the oxygen concentration sensor detects the concentration of oxygen in the gas environment inside the tank.
  • the solenoid valve is automatically opened, so that O 2 enters the tank from an external source of O 2 gas (for example, a cylinder in which O 2 is stored).
  • the solenoid valve closes.
  • the gas exchange between the small cell culture device and the bioreactor is to achieve gas exchange between the inside and the outside of the container through the venting gap between the container lid and the open edge of the container.
  • gas exchange between the inside and outside of the container is achieved by a microporous permeable membrane on the lid of the container.
  • gas exchange inside and outside the bag is achieved by gas diffusion of its own gas permeable, water-impermeable film.
  • a reactive ventilation type (or active ventilation type) ventilation method that is, by applying a positive pressure, a fresh gas containing a high oxygen content is input into the incubator and the reactor, and at the same time, a high carbon dioxide-containing gas is discharged. Metabolic gases.
  • a soft film bag bioreactor eg, a waved soft bag bioreactor
  • active ventilation is required regardless of size, for example, by using a gas blasted from the bottom of an airlift bioreactor to promote mixing of the reaction fluids to achieve gas exchange and nutrients. Pass.
  • Existing active aerated cell culture devices such as multi-layer cell culture devices - cell factories
  • bioreactors such as stirred tank reactors, soft membrane reactors, gas lift reactors, etc.
  • gases from external sources such as compressed gas in a gas cylinder, or gas produced by a gas compressor
  • filtered through a filter measuring and mixing different proportions of gas components, and then positive pressure into the cell culture or bioreactor, while the cell culture or The internal gas of the bioreactor is discharged.
  • Existing active ventilation is characterized by full gas filtration and transient drainage.
  • a sterilizing filter is also provided at the outlet end.
  • a positive pressure gas source, a gas inlet nozzle end and an outlet gas port end are required to be provided with a dead end filter type sterilization filter.
  • Existing active ventilation systems require O 2 and CO 2 sensors and gas mixers, pressure controllers, and flow controllers to ensure accurate gas ratios.
  • Existing active ventilation systems also require a heater at the gas discharge port to prevent the filter membrane from clogging the large amount of water vapor-exhausted gas. Therefore, the existing cell culture and bioreactor active ventilation systems are not only expensive but also complicated to operate, and in particular, are not suitable for biological fermentation and cell culture on a small scale (for example, an incubator scale).
  • the continuous transient exhaust gas also causes waste of clean gas and loss of the filter.
  • the invention utilizes existing equipment (for example, a carbon dioxide incubator, a three-gas incubator) to achieve precise regulation of the gas ratio, and uses a peristaltic pump to achieve non-invasive, non-contaminated transport of the gas, provides an active ventilation device and has the active ventilation
  • existing equipment for example, a carbon dioxide incubator, a three-gas incubator
  • a peristaltic pump to achieve non-invasive, non-contaminated transport of the gas
  • the device's active aerated bioreactors and cell culturers address the problems of expensive and complex gas regulation systems and complex operations in existing transient venting reactive venting bioreactors and cell culturers.
  • the present invention provides a living ventilator for a bioreactor and a cell culturer, and a cell culture and bioreactor using the bioreactor and the active ventilator for the cell culture device.
  • the invention utilizes existing equipment (for example, a carbon dioxide incubator, a three-gas incubator) to achieve precise regulation of the gas ratio, and utilizes a peristaltic pump to achieve gas-free transportation of the gas, thereby eliminating the need for a separate and complicated expensive gas filter, CO. 2 and O 2 sensors and controllers, gas mixers, pressure and flow control systems.
  • the active ventilating assembly of the present invention and the active aerated cell culture and bioreactor utilizing the active venting assembly are capable of saving gas and do not require a gas flow control system.
  • the invention provides a reactive venting assembly for a bioreactor and cell culture.
  • the reactive venting assembly includes one or more gas exchangers, each of the one or more gas exchangers having an interior space and configured to be disposed between the interior space and an exterior space outside of the interior space a gas exchange; and a line that is hermetically coupled between the one or more vessels and the one or more gas exchangers to form a gas flow loop, wherein the one or more vessels are provided with a living being Fermentation or cell culture materials and/or products.
  • At least a portion of the line is configured to be adapted to be driven by a gas drive such that gas flows in the gas flow circuit in a predetermined gas flow direction.
  • the gas passes sequentially through the gas drive, the vessel, the one or more gas exchangers, and back to the gas drive in the predetermined gas flow direction.
  • the active venting assembly of the present invention further includes one or more interface devices.
  • the one or more interface devices are configured to be hermetically coupled to the one or more containers, respectively.
  • the interface device includes at least two openings, two of the at least two openings being hermetically coupled to the pipeline, respectively.
  • the interface device may also include an opening that allows the biological fermentation or cell culture material and/or product to enter and/or remove the container. The opening remains hermetically sealed during the absence of the biological material entering and/or removing the container.
  • the interface device may include an intake pipe and an air outlet pipe extending into the interior of the container, the intake pipe and the air outlet pipe being hermetically coupled to the pipeline, respectively, and wherein the intake pipe extends into the interior of the container
  • the length of the outlet pipe extending into the interior of the container is different.
  • the length of the inlet tube extending into the interior of the container is greater than the length of the outlet tube extending into the interior of the container.
  • the portion of the intake tube that extends into the interior of the container includes a flexible tube that is provided with one or more through holes to allow gas to escape from the through hole.
  • the line can be hermetically coupled directly to the vessel to form the gas flow circuit.
  • At least one of the one or more gas exchangers can include one or more membranes.
  • the membrane includes a plurality of apertures configured to allow the gas to exchange and prevent passage of bacteria.
  • the at least one of the one or more gas exchangers may have a frame and the one or more membranes cover at least a portion of the frame.
  • the at least one of the one or more gas exchangers may further include at least one protective layer disposed outside the membrane.
  • the one or more gas exchangers can include a plurality of gas exchangers that are connected in parallel or in series with each other through the lines.
  • at least one of the one or more gas exchangers has a gas inlet and a gas outlet, wherein an air flow passage is defined between the gas inlet and the gas outlet, the direction of the gas passage being opposite to the preset The gas flows in a uniform direction, and wherein the gas inlet and the gas outlet are each hermetically coupled to the pipeline.
  • the gas inlet and the gas outlet may be disposed adjacent to each other.
  • the gas inlet and the gas outlet may be disposed away from each other.
  • the predetermined gas flow direction and the direction of gas exchange are substantially orthogonal to each other.
  • the gas drive device can include a peristaltic pump configured to alternately squeeze and relax the at least a portion of the line such that gas is along the gas flow circuit The flow of the preset gas flows.
  • the reactive vent assembly can also include a gas pressure regulating device disposed in the gas flow circuit and hermetically coupled to the line.
  • the air pressure adjusting device may be disposed between the container and the one or more gas exchangers. The air pressure adjusting device can be configured to change and /
  • the invention provides an active gas circulation system comprising an active venting assembly as provided in one aspect of the invention, and the one or more containers.
  • the container may comprise one or more of a bottle, a soft bag or a cell factory.
  • the gas circulation system also includes one or more interface devices.
  • the one or more interface devices are configured to be hermetically coupled to the one or more containers, respectively.
  • the interface device includes at least two openings, two of the at least two openings being hermetically coupled to the pipeline, respectively.
  • the interface device also includes an opening that allows the biological fermentation or cell culture material and/or product to enter and/or remove the container. The opening remains hermetically sealed during the absence of the biological material entering and/or removing the container.
  • the interface device may include an intake pipe and an air outlet pipe that extend into the interior of the container, the intake and exhaust pipes being hermetically coupled to the pipeline, respectively. The length of the inlet tube extending into the interior of the container is different than the length at which the outlet tube extends into the interior of the container.
  • the length of the inlet tube extending into the interior of the container may be greater than the length of the outlet tube extending into the interior of the container.
  • the portion of the intake tube that extends into the interior of the container includes a flexible tube that is provided with one or more through holes to allow gas to escape from the through hole.
  • the line can be hermetically coupled directly to the vessel to form the gas flow circuit.
  • the container may also be provided with openings for allowing the bio-fermentation or cell culture material and/or product to enter and/or remove the container.
  • At least one of the one or more gas exchangers includes one or more membranes.
  • the membrane includes a plurality of apertures configured to allow the gas to exchange and prevent passage of bacteria.
  • the at least one of the one or more gas exchangers can have a frame, and wherein the one or more membranes cover at least a portion of the frame.
  • the at least one of the one or more gas exchangers may further include at least one protective layer disposed outside the membrane.
  • the one or more gas exchangers comprise a plurality of gas exchangers that are connected in parallel or in series with one another via the lines.
  • at least one of the one or more gas exchangers has a gas inlet and a gas outlet.
  • An air flow passage is defined between the gas inlet and the gas outlet, the direction of the gas passage being coincident with the predetermined gas flow direction.
  • the gas inlet and the gas outlet are each hermetically coupled to the line.
  • the gas inlet and the gas outlet may be disposed adjacent to each other or away from each other.
  • the predetermined gas flow direction and the direction of the gas exchange are substantially orthogonal to each other.
  • the gas drive can include a peristaltic pump.
  • the peristaltic pump is configured to alternately compress and relax the at least a portion of the line such that gas flows in the gas flow circuit along the predetermined gas flow direction.
  • the gas circulation system further includes a gas pressure regulating device disposed in the gas flow circuit and hermetically coupled to the gas line.
  • the air pressure adjusting device may be disposed between the container and the one or more gas exchangers.
  • the gas pressure regulating device can be configured to change and/or maintain the diameter of the line to change and/or maintain the pressure of the gas within the line.
  • the gas circulation system also includes a controller.
  • the controller is communicatively coupled to the gas drive and configured to adjust operation of the gas drive based on pressure in the gas flow circuit.
  • the pressure in the gas flow circuit can be measured at the line or the vessel.
  • the pressure in the gas flow circuit can be measured in a non-contact manner.
  • the one or more containers include a plurality of containers that are connected to each other in parallel or in series by the pipeline.
  • the invention provides an active venting assembly.
  • the active venting assembly includes at least three components in communication with an internal gas flow passage: a gas exchanger, a peristaltic pump tubing, and an airway.
  • the gas exchanger is a cavity having an internal gas flow passage and is connected in series between the two pipe sections of the air pipe, and at least one wall surface of the cavity is sealed to allow gas to permeate to realize gas exchange inside and outside the cavity but block bacteria Through the microporous permeable membrane.
  • the peristaltic pump tube is a peristaltic pump head tube card and an elastic hose tube section that squeezes and drives the internal gas flow.
  • the air conduit is a gas transmission conduit that communicates between the peristaltic pump tube and the gas exchanger and between the intake and outlet tubes of the bioreactor or cell culture device to be used, respectively.
  • the gas exchanger is a microporous gas permeable filter bag cavity having an internal gas flow passage fused from two layers of the microporous gas permeable membrane. Both ends of the microporous permeable filter bag are fused with a connecting nozzle. The connecting nozzle and the air guiding tube are connected by a tight fit or an integrated fusion.
  • the gas exchanger is a microporous gas permeable filter bag cavity having an internal gas flow passage fused from two layers of the microporous gas permeable membrane.
  • One end of the microporous gas permeable filter bag is fused with two communicating nozzles, and two soft films between the two communicating nozzles are fused to the bag to form a U-shaped air flow channel communicating with the two communicating nozzles.
  • the connecting nozzle and the air guiding tube are connected by a tight fit or an integrated fusion.
  • the gas exchanger is a microporous gas permeable bag cavity having an internal gas flow passage fused by a two-layer barrier film provided with a gas permeable membrane sealed venting window.
  • the microporous permeable filter bag is fused with a connecting nozzle at both ends, and the connecting nozzle and the air guiding tube are connected by a tight fit or an integrated fusion.
  • the gas exchanger is a microporous gas permeable bag cavity having an internal gas flow passage fused by a two-layer barrier film provided with a gas permeable membrane sealed venting window.
  • One end of the microporous permeable filter bag is fused with two connecting nozzles.
  • the two layers of soft film between the two communicating nozzles extend into the pocket to form a U-shaped air flow passage that communicates with the two communicating nozzles.
  • the connecting nozzle and the air guiding tube are connected by a tight fit or an integrated fusion.
  • the gas exchanger is a rigid frame cavity formed by sealing a microporous gas permeable membrane on a support frame of a hard material having a connection nozzle at both ends.
  • the communication nozzle and the air guiding tube are connected by a close fit.
  • a mesh sheet with a vent mesh to support the protection of the microporous permeable membrane is attached to the outside of the microporous membrane.
  • the air conduits connect two or more gas exchangers in parallel or in series through a three-way joint or a multi-way joint to increase gas exchange efficiency, or to connect two or more bioreactors or cells in parallel or in series.
  • the incubator is used to increase the size of the culture.
  • the airway tube is disposed on a pipe section between the gas exchanger and the gas pipe of the bioreactor or cell culture device to adjust the bioreactor or cell culture device by adjusting the size of the pipe diameter.
  • Air pressure regulator for internal air pressure.
  • the reactive aerated bioreactor is configured with an active venting assembly provided by one aspect of the invention.
  • the bioreactor is a wave bioreactor, an airlift bioreactor, a shake flask bioreactor or a stirred bioreactor.
  • the bioreactor is provided with an intake pipe and an outlet pipe for gas to enter and exit.
  • the air duct of the active ventilating assembly communicates with the intake and outlet tubes of the bioreactor to form a closed loop of the air flow passage.
  • the bioreactor is a soft membrane bioreactor.
  • the soft membrane bioreactor is a planar (2D) soft membrane bioreactor or a three-dimensional (3D) soft membrane bioreactor fused by two layers of soft membrane.
  • the three-dimensional soft membrane bioreactor has a surface area of at least one soft film of the two soft films in the fusion ring which is larger than a plane area enclosed by the fusion ring by 5% or more.
  • the inlet and outlet tubes of the soft membrane bioreactor are fused to the fusion side of the two layers of the soft membrane reactor, or are fused to the planar soft membrane, or are fused to
  • the nozzle of the soft film bag is placed on the nozzle cover.
  • the intake pipe and the outlet pipe that are fused to the nozzle cover are respectively disposed on different nozzle covers or the same nozzle cover.
  • the intake and outlet tubes disposed in the same nozzle cover extend into the nozzle or at different depths within the reactor.
  • the soft membrane bioreactor is a waved soft membrane bioreactor that achieves liquid mixing and gas exchange by shaking the internal liquid to form a wave.
  • the soft membrane bioreactor is an airlift type soft membrane bioreactor that achieves liquid mixing and gas exchange by aeration to a reaction liquid inside thereof.
  • the air intake pipe of the airlift type soft membrane bioreactor is connected in the bag with an aeration pipe extending to a small aeration hole at the bottom of the reactor.
  • the reactive aerated cell culture device is configured with an active venting assembly provided in accordance with an aspect of the invention.
  • the cell culture device is a single-layer cell culture vessel-culture flask or a multi-layered rigid plastic cell culture vessel-cell factory.
  • the single or multi-layered rigid plastic cell culture device has one or more incubator nozzles.
  • a nozzle cover is disposed on the mouth of the incubator.
  • An intake pipe or/and an air outlet pipe are disposed through the nozzle cover.
  • the intake pipe and the outlet pipe may be disposed on the same nozzle cover or respectively disposed on different nozzle covers.
  • the intake and outlet tubes disposed in the same nozzle cover extend into the nozzle or at different depths within the reactor.
  • the air duct of the active ventilator assembly is in communication with the inlet and outlet tubes of the cell culture device to form a closed loop of the air flow passage.
  • FIG. 1 is a schematic structural view of an exemplary embodiment of an active ventilation assembly of the present invention.
  • FIG. 2 is a schematic structural view of still another exemplary embodiment of the active ventilation assembly of the present invention.
  • FIG. 3 is a schematic illustration of the structure of an embodiment of a bioreactor or cell culture device having an active venting assembly of the present invention.
  • FIG. 4 is a schematic structural view of an exemplary embodiment of a gas exchanger of the present invention.
  • Fig. 5 is a schematic structural view of still another exemplary embodiment of the gas exchanger of the present invention.
  • Fig. 6 is a schematic structural view of still another exemplary embodiment of the gas exchanger of the present invention.
  • Fig. 7 is a schematic structural view of still another exemplary embodiment of the gas exchanger of the present invention.
  • Figure 8 is a schematic structural view of still another exemplary embodiment of the gas exchanger of the present invention.
  • Figure 9 is a schematic structural view of still another exemplary embodiment of the gas exchanger of the present invention.
  • Figure 10 is a schematic view showing the structure of still another exemplary embodiment of the gas exchanger of the present invention.
  • Figure 11 is a block diagram showing an exemplary embodiment of an interface device of the present invention.
  • Figure 12 is a schematic illustration of an exemplary embodiment of the use of the bioreactor of the present invention.
  • Figure 13 is a schematic view showing the structure of still another embodiment of the bioreactor of the present invention.
  • Figure 14 is a schematic illustration of the use of an exemplary embodiment of the use of the bioreactor of the present invention.
  • Figure 15 is a schematic view showing the structure of still another embodiment of the bioreactor of the present invention.
  • Figure 16 is a schematic view showing the structure of still another embodiment of the bioreactor of the present invention.
  • Figure 17 is a schematic view showing the structure of still another embodiment of the bioreactor of the present invention.
  • Figure 18 is a schematic structural view of an exemplary embodiment of a cell culture device of the present invention.
  • Figure 19 is a schematic view showing the structure of still another exemplary embodiment of the cell culture device of the present invention.
  • Figure 20 is a schematic view showing the state of use of the living bioreactor and the cell culture device of the present invention.
  • the present invention provides a reactive venting assembly for bioreactors and cell cultures that is suitable for use with existing bioreactors or cell culture vessels.
  • the active venting assembly can provide a closed gas flow circuit after being coupled to the bioreactor or cell culture device.
  • the invention also provides a bioreactor comprising a reactive venting assembly and a cell culture device.
  • At least the gas exchanger of the active venting module, bioreactor or cell culture device of the present invention can be placed in a carbon dioxide incubator or a three gas incubator for gas exchange.
  • the sensor system and gas composition setting and holding system of the carbon dioxide incubator or the three-gas incubator can ensure the stability of the gas environment in the carbon dioxide incubator or the three-gas incubator.
  • the active gas exchange module of the present invention can realize active gas exchange between the internal gas of the bioreactor or the cell culture device and the gas environment in the carbon dioxide incubator or the three gas incubator, thereby ensuring the bioreactor and cell culture.
  • the gas inside the device maintains the desired composition.
  • the active ventilating assembly provided by the present invention and the bioreactor and cell culture device using the active venting assembly achieve a cyclic gas exchange.
  • the exchange of gas components (such as O 2 and CO 2 ) inside and outside the cell culture or bioreactor is achieved by a gas exchanger in the active venting assembly, thereby saving gas and greatly reducing the consumption of the gas source. Thanks to the use of existing bio incubators for precise control of the temperature, humidity and gas configuration of the gaseous environment, there is no need to set up expensive gas sensing equipment or complex pressure, flow control systems and gas filtration systems.
  • the culture solution in the cell culture or bioreactor does not evaporate and does not clog the filter.
  • FIG. 1 is a schematic view showing the structure of an exemplary embodiment of a living ventilating assembly for a bioreactor and a cell culture device of the present invention.
  • the active venting assembly includes a gas exchanger 1 and an airway tube 3 (ie, a line system).
  • the gas exchanger 1 and the air guiding tube 3 are hermetically coupled.
  • the airway tube can include multiple segments instead of having to be a continuous line. For example, one segment of the air conduit can be hermetically coupled to the intake end of the gas exchanger, while the other segment can be hermetically coupled to the gas outlet of the gas exchanger, thereby allowing the gas conduit and gas exchanger
  • the gas flow path is provided as a whole.
  • the airway tube 3 can include a peristaltic pump tube 2.
  • the peristaltic pump tube 2 comprises an elastic hose adapted to be squeezed by, for example, a peristaltic pump head such that gas flows inside the peristaltic pump tube.
  • the peristaltic pump tube 2 and other portions of the airway tube 3 may be made of the same or different materials.
  • the material of the peristaltic pump tube has a certain elasticity.
  • the material of the peristaltic pump tube has the characteristics of wear resistance, pressure bearing capacity, hardness, airtightness, low adsorption, high temperature resistance, aging resistance, non-swelling, corrosion resistance, and low precipitation.
  • the material of the peristaltic pump tube may include silicone rubber, fluororubber, Teflon, rubber, plastic, synthetic materials, and the like.
  • the peristaltic pump tubing 2 and other portions of the airway tube 3 can be communicated through a connector 34 such that the peristaltic pump tubing 2 is replaceable.
  • FIG. 2 is a schematic view showing the structure of still another exemplary embodiment of the active ventilation assembly of the present invention.
  • the peristaltic pump tubing 2 and the other portions of the airway tube 3 are integrally connected.
  • the peristaltic pump tube 2 and other portions of the airway tube 3 may be made of the same material, for example, the peristaltic pump tube 2 and other portions of the air tube 3 are also elastic hoses.
  • the elastic hose section clamped by the pump head of the peristaltic pump is a peristaltic pump pump tube.
  • the gas exchanger 1 is coupled in series with the gas conduit 3.
  • the gas exchanger 1 has an internal space in which an internal gas flow passage is realized.
  • the gas exchanger 1 is configured to effect gas exchange between the internal space and an external space outside the internal space.
  • the gas exchanger 1 is configured to prevent bacteria and dust from the external space from entering the interior space when gas exchange is achieved.
  • the gas exchanger can include a microporous gas permeable membrane.
  • the microporous gas permeable membrane has a plurality of micropores sized to block the passage of bacteria and dust while allowing free passage of gas.
  • the gas flow direction within the gas exchanger may be substantially perpendicular to the microporous gas permeable membrane.
  • the gas exchange of the gas component through the microporous gas permeable membrane is achieved by the difference in partial pressure caused by the difference between the gas component inside the gas exchanger 1 and the gas component outside the gas exchanger 1.
  • This gas exchange ensures that the gas component inside the gas exchanger 1 coincides with the external gas component. For example, if the CO 2 gas content of CO 2 gas content of the gas exchanger of the internal gaseous environment a gas exchanger is greater than 1 atmosphere outside, the inside of the gas exchanger 1 CO 2 gas through the microporous breathable The filter membrane is discharged to the outside of the gas exchanger 1.
  • the external gas exchange of the O 2 gas permeable microporous breathable 1 enters the inside of the gas exchanger 1. In this way, the composition of the gas continuously flowing through the inner space of the gas exchanger 1 via the air conduit can be made to coincide with the composition of the gaseous environment of the outer space of the gas exchanger 1.
  • the internal exchange of the gas and the external gas of the gas exchanger can be achieved by increasing the surface area of the microporous membrane of the gas exchanger. It is also possible to achieve a full exchange of gases by providing a plurality of gas exchangers, as described in detail below.
  • the microporous gas permeable membrane may be a hydrophobic microporous membrane, such as a polytetrafluoroethylene PTFE hydrophobic vent membrane.
  • the pore size of the pores on the hydrophobic microporous membrane may be 0.01 ⁇ m, 0.02 ⁇ m, 0.05 ⁇ m, 0.07 ⁇ m, 0.09 ⁇ m, 0.1 ⁇ m, 0.22 ⁇ m, 0.3 ⁇ m, 0.45 ⁇ m or a value between any two of the above values.
  • the microporous gas permeable membrane can have the properties of gas permeability, water impermeability, flame retardancy, high temperature resistance, strong acid and alkali resistance, non-toxicity and the like.
  • the active ventilation assembly can also include an interface device.
  • the interface device can be hermetically coupled to the container that mates with the active venting assembly, as described in detail below.
  • FIG 3 is a schematic illustration of the construction of an embodiment of a bioreactor or cell culture device having an active venting assembly of the present invention.
  • the container 5 can be a bioreactor or cell culturer of any shape, volume or material, such as a bottle or bag.
  • the container 5 can be a planar soft film wave bioreactor.
  • the planar soft-film wave bioreactor is formed by two layers of planar soft membranes, on which the reactor nozzles are fused, and the two membranes are fused at the periphery to form a fusion ring 32.
  • the interface device 11 can include a nozzle cover that mates with the open end of the container.
  • the spout can be threaded or otherwise airtightly coupled to the opening of the container.
  • the spout can be a plug that can be airtightly inserted into the opening of the container.
  • the interface device 11 is provided with an opening 12 and an opening 13 at least penetratingly.
  • the opening 12 and the opening 13 are hermetically coupled to the air duct, respectively.
  • the opening 12 can be an air inlet.
  • the opening 13 can be an air outlet.
  • the inlet device 12 and the outlet pipe 13 may be provided on the interface device 11.
  • the intake pipe 12 and the air outlet pipe 13 are airtightly coupled to the air guide tubes, respectively.
  • the interface device 11 is also provided with an opening for the biological sample to enter and/or exit the bioreactor 5.
  • the air conduit 3 is coupled to the interface device 11 and the gas exchanger 1 to form a gas flow circuit from the interface device 11 to the gas exchanger 1 to the interface device 11.
  • At least a portion of the airway tube 3 is configured to be adapted to be driven by a gas drive such that gas flows in the gas flow circuit.
  • the gas drive can be a peristaltic pump or a compressor.
  • the peristaltic pump tube 2 of the airway tube 3 can be embedded in the card slot of the peristaltic pump head 6.
  • the motor driver 17 drives the rotor 18 of the peristaltic pump to rotate.
  • a roller 19 is disposed on the rotor 18. The rotation of the rotor 18 drives the roller 19 to repeatedly press the peristaltic pump pump tube 2, thereby driving the gas inside the peristaltic pump tube 2 to flow into the container through the air guiding tube 3.
  • a gas pressure regulator 4 may also be provided on the air conduit 3 between the gas exchanger 1 and the gas outlet 13.
  • the gas pressure regulator 4 is configured to maintain the bulging state of the bioreactor 5 and adjust the internal gas pressure of the bioreactor 5 by adjusting the inner diameter of the air tube 3.
  • the gas pressure regulator may be provided with a pressure sensor for measuring the gas pressure in the air guiding tube. The operation of the gas drive can be controlled based on the measured gas pressure. For example, if the measured gas pressure is less than a preset value, the increase in the rotational speed of the peristaltic pump can be controlled. It is also possible to measure the gas pressure in the vessel or in the pipeline in a non-contact manner.
  • a non-contact pressure measuring device can be provided to measure the gas pressure in the planar soft membrane bioreactor by injecting gas onto the surface of the planar soft membrane bioreactor and measuring the pressure of the reflected gas.
  • the gas pressure in the vessel is substantially consistent with the gas pressure within the airway.
  • a circulating gas flow circuit is formed in the active venting assembly and vessel.
  • the gas circulates in a predetermined direction in the gas flow circuit.
  • the gas proceeds from the vessel along the predetermined gas flow direction, passes through the gas exchanger, the gas drive, and returns to the vessel.
  • the gas exchanger may be disposed downstream of the vessel such that gas driven by the gas drive is first fed to the vessel and gas exiting the vessel enters the gas exchanger for component exchange.
  • the container may be separate from the gas delivery device.
  • the gas delivery device of the present invention can be used in conjunction with commercially available containers.
  • both ends of the gas delivery tube of the gas delivery device of the present invention can be inserted into the lid of the container to effect gas communication between the gas delivery device and the container.
  • the gas delivery device of the present invention can have an interface device.
  • the interface device is an adapter that mates with the container, such as a plug or a screw cap. Both ends of the gas delivery tube of the gas delivery device can be hermetically coupled to the interface device.
  • the gas delivery device effects gas communication with the container by hermetically coupling the interface device to the container.
  • the container may be provided integrally with a gas delivery device.
  • the two ends of the gas delivery tube of the gas delivery device are integrally coupled to the container to effect gas communication with the container.
  • an opening for the entry/exit of biological material can also be provided.
  • biological materials can include cells, bacteria, fungi, or organisms.
  • the gas exchanger 1 includes a support frame 8 and a membrane 9 disposed on opposite sides of the support frame 8.
  • the support frame 8 can be in the shape of a flat box.
  • the support frame 8 can be integrally thermoformed.
  • the support frame 8 has a connection nozzle 7 at both longitudinal ends, and the air guide tube 3 is coupled to the connection nozzle 7.
  • the two connecting nozzles 7 define an internal gas flow passage of the gas exchanger 1, which is identical to the gas flow circuit described above.
  • the internal gas flow passage is substantially perpendicular to the membrane.
  • the membrane 9 may be a microporous permeable membrane. The area of the microporous gas permeable membrane can be matched to the flow rate and/or flow rate of the gas flowing through the internal gas flow path of the gas exchanger 1, thereby achieving sufficient gas exchange between the interior and exterior of the gas exchanger.
  • a microporous gas permeable membrane having an increased area may be provided such that gas flowing through the internal gas flow path of the gas exchanger 1 can be fully exchanged with the gas environment outside the gas exchanger 1, achieving gas
  • the gas composition inside and outside of the exchanger 1 is identical.
  • a plurality of gas exchangers 1 may be provided to achieve sufficient exchange of gas flowing through the internal gas flow passages of the gas exchanger 1 with the external gaseous environment.
  • the plurality of gas exchangers 1 may be connected to each other in series via a vent pipe or may be connected in parallel to each other.
  • the vent tube can be provided with three interfaces to achieve a parallel connection of the three gas exchangers.
  • the gas exchanger 1 is further provided with a mesh sheet 10 provided on the microporous gas permeable membrane 9.
  • the mesh sheet 10 may be a gas permeable protective film.
  • the mesh sheet 10 can support and protect the microporous gas permeable membrane 9.
  • the gas exchanger 1 includes a support frame 8.
  • the support frame 8 may have a cylindrical shape.
  • the support frame 8 can be integrally thermoformed.
  • a connecting nozzle 7 is disposed on the circumferential wall of the support frame, and the air guiding tube 3 is coupled to the connecting nozzle 7.
  • the two connecting nozzles 7 on the circumferential wall of the support frame define an internal gas flow passage of the gas exchanger 1, which is identical to the gas flow circuit described above.
  • the internal gas flow passage is substantially perpendicular to the membrane.
  • a microporous gas permeable membrane is sealingly disposed on the end surface of the tube of the support frame 8.
  • a mesh sheet 10 may also be provided on the microporous gas permeable membrane to support and protect the microporous membrane.
  • the gas exchanger 1 includes a support frame 8.
  • the support frame 8 may have a cylindrical shape.
  • the support frame 8 can be an integral thermoplastic.
  • a connecting nozzle 7 is disposed on the end wall of the support frame, and the air guiding tube 3 is coupled to the connecting nozzle 7.
  • the two connecting nozzles 7 on the barrel end wall of the support frame define an internal gas flow passage of the gas exchanger 1, the internal gas flow passage being identical to the gas flow circuit described above.
  • the internal gas flow passage is substantially perpendicular to the membrane.
  • a microporous gas permeable membrane is sealingly disposed on the circumferential surface of the support frame 8.
  • a mesh sheet 10 may also be provided on the microporous gas permeable membrane to support and protect the microporous membrane.
  • the gas exchanger may not have a support frame.
  • the gas exchanger 1 may be a microporous gas permeable membrane bag made of two of the microporous gas permeable membranes 9. The edges of the two microporous permeable membranes 9 can be thermocompression bonded together to form the pouch.
  • the microporous gas permeable membrane bag may be rectangular, circular, elliptical or any other shape.
  • a communication nozzle 7 may be disposed on both longitudinally opposite sides of the microporous gas permeable membrane bag.
  • the two connecting nozzles 7 define an internal gas flow passage of the gas exchanger 1, which is identical to the gas flow circuit described above.
  • the internal gas flow passage is substantially perpendicular to the microporous gas permeable membrane 9.
  • the gas exchanger 1 is a microporous gas permeable membrane bag.
  • the microporous gas permeable membrane bag may not have a support frame.
  • the microporous gas permeable membrane bag may comprise two soft membranes with a window. The edges of the two pellicles can be thermocompression bonded together to form the pouch.
  • the bag can have any suitable shape and geometry.
  • the window region of the soft film may be circular, rectangular, elliptical or any other suitable shape.
  • a microporous gas permeable membrane 9 is provided in the window region of the soft film. The microporous gas permeable membrane 9 can be pressed to the inner edge of the window region of the soft film.
  • a fusion edge 33 may be formed in an overlapping region of the inner edge of the window region of the soft film and the microporous gas permeable membrane 9.
  • a communication nozzle 7 may be disposed on both longitudinally opposite sides of the microporous gas permeable membrane bag.
  • the two connecting nozzles 7 define an internal gas flow passage of the gas exchanger 1, which is identical to the gas flow circuit described above.
  • the internal gas flow passage is substantially perpendicular to the microporous gas permeable membrane 9.
  • the window region of the soft film is U-shaped, and the microporous permeable membrane is Two communication nozzles 7 are provided on the same side of the bag such that the two communication nozzles 7 are adjacent to each other.
  • the two communication nozzles 7 are respectively in communication with the internal air flow passages in the U-shaped microporous gas permeable membrane 9.
  • the internal air flow passage of the gas exchanger 1 is formed in a U shape.
  • the U-shaped internal gas flow passage increases the length of the internal gas flow passage and enhances the gas exchange effect of the gas passing through the microporous gas permeable membrane.
  • the internal gas flow passage is substantially perpendicular to the microporous gas permeable membrane 9. .
  • FIG 11 is a block diagram showing an exemplary embodiment of an interface device of the present invention.
  • the interface device 11 can be a nozzle cover that is coupled to the container.
  • the nozzle cover can be a plug.
  • a plug ring may be provided on the plug to cooperate with the open end of the bioreactor to achieve a gas-tight coupling with the container.
  • the spout can be a threaded cap.
  • the threaded cap may be provided with an external thread that cooperates with the internal thread of the open end of the bioreactor to achieve a gas-tight coupling with the container.
  • An intake pipe 12 and an air outlet pipe 13 are provided through the interface device 11.
  • the length of the intake pipe 12 and the air outlet pipe 13 extending within the nozzle cover may be different to prevent a short circuit of the gas from occurring. That is, the gas that has entered the container from the intake pipe 12 is prevented from being discharged from the air outlet pipe 13 without being sufficiently mixed with the gas in the container.
  • the depth of the air intake tube 12 extending within the nozzle cover may be greater than the length of the air outlet tube 13 extending within the nozzle cover.
  • the intake manifold 12 can extend below the level of liquid in the bioreactor such that the gas pumped by the peristaltic pump is sufficiently in contact with the liquid within the bioreactor.
  • a plurality of nozzle covers 11 may be provided, and the intake pipe 12 and the air outlet pipe 13 may be respectively disposed on two different nozzle covers.
  • An opening for the biomaterial to enter/exit the bioreactor may also be provided on the spout lid 11.
  • FIG 12 is a schematic illustration of an exemplary embodiment of the use of a bioreactor of the present invention.
  • the waved soft membrane bioreactor of the present invention e.g., the bioreactor shown in Figure 1
  • the membrane bioreactor is fixed in the fixed frame clamp 14 and then placed on the platform 15 of the shaker 16.
  • the peristaltic pump head 6 can be mounted to the peristaltic pump actuator of the shaker and the peristaltic pump tubing 2 can be snapped into the slot of the peristaltic pump head 6.
  • the shaking of the shaker platform 15 drives the culture liquid inside the soft membrane bioreactor to form a wave, thereby achieving full exchange and mixing of the gas and the liquid in the soft membrane bioreactor.
  • the gas inside the membrane bioreactor is pumped into the gas exchanger 1 under the drive of a peristaltic pump.
  • the gas in the inner space of the gas exchanger 1 and the external gas atmosphere in the outer space of the gas exchanger 1 realize gas exchange across the microporous gas permeable membrane 9 under the action of the partial pressure difference.
  • the high oxygen partial pressure O 2 outside the gas exchanger 1 enters the gas exchanger 1, and the high carbon dioxide partial pressure CO 2 in the gas exchanger 1 is dispersed outside the gas exchanger 1.
  • a gas pressure regulator 4 may also be disposed on the air conduit 3 between the gas exchanger 1 and the gas outlet tube 13. The gas pressure regulator 4 can adjust the inner diameter of the air guiding tube 3 to adjust the bulging state and internal pressure of the soft membrane bioreactor.
  • the shaker, soft membrane bioreactor and gas exchanger can be placed in a carbon dioxide incubator or a three gas incubator.
  • the gas control system of the incubator can be used to accurately control and maintain the temperature, humidity, pressure and composition of the gas environment in the incubator. Further, precise exchange of oxygen and carbon dioxide components in the bioreactor is achieved by gas exchange of the gas exchanger 1.
  • the shaker 16 can be divided into a plane track shaker, a three-dimensional shaker and a seesaw shaker according to the manner in which it is shaken.
  • a suitable shaker can be selected depending on the type of cells to be cultured, the structure of the bioreactor, the mixing effect requirements, and the shearing force requirements.
  • the cell culture device is placed in a carbon dioxide incubator or a three-gas incubator.
  • two or more gas exchangers 1 may be connected in series or in parallel through the air duct 3 using a three-way or multi-way joint to increase the gas exchange area. , thereby increasing gas exchange efficiency.
  • two or more bioreactors may be connected in series or in parallel through the airway tube 3 using a three-way or multi-pass joint.
  • FIG. 13 is a schematic view showing the structure of still another embodiment of the bioreactor of the present invention.
  • the intake pipe 12 and the outlet pipe 13 of the soft membrane bioreactor in the present embodiment are integrally provided on the edge of the soft membrane bioreactor.
  • the soft membrane bioreactor can be a planar soft membrane wave bioreactor 20 made by hot pressing two sheets of film.
  • the intake pipe 12 and the air outlet pipe 13 are disposed at the fusion edges of the two sheets of film.
  • the intake pipe 12 and the outlet pipe 13 may be disposed adjacent to each other on the bioreactor 20.
  • the bioreactor 20 may also include a liquid inlet and outlet tube 23 for injecting liquid material and/or biological material into the bioreactor 20 and withdrawing liquid material and/or biological material from the bioreactor 20.
  • FIG 14 is a schematic illustration of the use of an exemplary embodiment of the use of a bioreactor of the present invention.
  • the membrane bioreactor 20 can be secured with a retaining clip 14 and placed obliquely on the shaker platform 15 by an L-shaped bracket 27. Such a structure prevents the culture liquid from entering the gas outlet pipe 13 during the shaking of the shaker.
  • the soft membrane bioreactor 20 can be an infusion bag that is widely used in the medical field.
  • the active ventilating assembly of the present invention can be coupled to an infusion bag to form a vented membrane bioreactor.
  • the infusion bag is a commonly used medical product, which has the advantages of non-toxic and harmless, no heat source, good sealing, no leakage, and sterility. Used in combination with the active ventilating assembly of the present invention, it can be used for suspension culture.
  • soft film bags similar in structure to the infusion bag such as blood transfusion bags and urine bags, can also be used.
  • FIG. 15 is a schematic view showing the structure of still another embodiment of the bioreactor of the present invention.
  • the bioreactor can be an airlift bioreactor.
  • the soft membrane bioreactor of this embodiment can be formed by hot pressing of two sheets of soft film structure and form a fusion ring 32.
  • An intake pipe 12 and an air outlet pipe 13 are provided on the nozzle cover 11. Outside the membrane bioreactor, a liquid inlet and outlet pipe 23 can be connected to the intake pipe 12 via a three-way joint 21.
  • an aeration tube 24 can be connected to the intake pipe 12.
  • the aeration tube 24 is provided with a plurality of micropores. The gas input through the intake pipe 12 can be discharged through the micro holes in the aeration pipe 24.
  • the aeration tube 24 can extend to the bottom of the bioreactor.
  • the gas pumped to the intake pipe 12 by the peristaltic pump enters the aeration tube 24, and microbubbles are ejected through the micropores.
  • the microbubbles rise in the liquid.
  • the liquid containing microbubbles has a lower density and rises, resulting in high-density liquid movement on both sides, thus forming a liquid up-and-down cycle in the liquid and achieving sufficient gas exchange. Therefore, the soft membrane bioreactor of the present embodiment can achieve liquid up and down circulation and full exchange of gas without shaking of the shaker.
  • FIG 16 is a schematic view showing the structure of still another embodiment of the bioreactor of the present invention.
  • the bioreactor can be a three dimensional soft membrane bioreactor.
  • a three-dimensional soft membrane reactor can be made by thermally pressing two soft membrane structures together.
  • the three-dimensional soft membrane reactor can be fabricated using a thermoplastic molding process in conjunction with a mold.
  • a fusion ring 32 is formed in a portion where the two soft films are thermally fused together.
  • One of the two sheets of soft film has a surface area greater than the other sheet.
  • the surface area of one of the two soft films is greater than 5% of the surface area of the other film, so that after injecting the liquid into the three-dimensional soft membrane bioreactor, the three-dimensional soft film organism
  • the reactor presents a three-dimensional construction.
  • the three-dimensional soft membrane bioreactor may be a three-dimensional soft membrane wave bioreactor 25, the upper soft membrane being a planar soft membrane, and the lower soft membrane being a hemispherical three-dimensional soft membrane.
  • a nozzle cover 11 is provided on the upper soft film.
  • a three-dimensional hard-shell carrier 26 that conforms to the three-dimensional configuration of the three-dimensional soft-film wave bioreactor 25 can be provided to support the three-dimensional soft-film wave bioreactor 25.
  • the three-dimensional hard shell holder 26 can be thermoformed from the hard sheet material using the same mold as the three-dimensional soft film wave bioreactor 25.
  • the structure of the three-dimensional soft membrane bioreactor of this exemplary embodiment is similar to a rigid inverted conical or hemispherical shake flask. Therefore, the three-dimensional soft membrane bioreactor can be shaken using a planar gyro shaker.
  • FIG 17 is a schematic view showing the structure of still another embodiment of the bioreactor of the present invention.
  • the bioreactor of this embodiment can be made by thermally pressing two sheets of the soft film structure together.
  • the three-dimensional soft membrane reactor can be fabricated using a thermoplastic molding process in conjunction with a mold.
  • both of the soft films have a three-dimensional configuration, thereby constituting a double-sided three-dimensional soft film wave bioreactor 35.
  • the intake pipe 12 and the air outlet pipe 13 and the liquid inlet and outlet pipe 23 may be integrally provided on the fusion ring 32.
  • a three-dimensional hard-shell carrier 26 that conforms to the three-dimensional configuration of the double-sided three-dimensional soft membrane reactor 25 can be provided.
  • a three-dimensional hard sheet carrier 26 is provided under the double-sided three-dimensional soft membrane reactor 25 to support the double-sided three-dimensional soft membrane reactor 25.
  • the three-dimensional hard shell holder 26 can be thermoformed from the hard sheet material using the same mold as the double-sided three-dimensional soft membrane reactor 25.
  • the three-dimensional hard shell holder 26 can be placed on the L-shaped bracket such that the double-sided three-dimensional soft-film wave bioreactor 25 is placed on the shaker platform in a tilted manner, thereby placing the liquid in the double-sided three-dimensional soft membrane reactor 25 into the chamber.
  • Outlet pipe 13 is provided under the double-sided three-dimensional soft membrane reactor 25 to support the double-sided three-dimensional soft membrane reactor 25.
  • the three-dimensional hard shell holder 26 can be thermoformed from the hard sheet material using the same mold as the double-sided three-dimensional soft membrane reactor 25.
  • the three-dimensional hard shell holder 26 can be placed on the L-shaped bracket such that the double-sided three-dimensional soft-film wave bioreactor 25
  • the active venting assembly of the present invention can be mated with a variety of different bioreactors.
  • the bioreactor is not limited to the species listed in the above exemplary embodiments.
  • the bioreactor can be a shake flask bioreactor, including a conical flask and a cone bottom bottle.
  • the bioreactor can also be a stirred tank, including a magnetically driven stirred tank and a mechanically driven stirred tank.
  • the active venting assembly of the present invention is equally applicable to cell culture vessels, although described using bioreactors in the above exemplary embodiments.
  • FIG 18 is a schematic view showing the structure of an exemplary embodiment of the cell culture device of the present invention.
  • the cell culture device of this embodiment may be a single layer cell culture flask 28.
  • the interface device 11 e.g., the nozzle cover
  • An intake pipe 12 and an air outlet pipe 13 are provided through the nozzle cover 11 so as to be continuous therethrough.
  • the length of extension of the intake tube 12 and the outlet tube 13 within the nozzle cover may vary.
  • the depth of the air intake tube 12 extending within the nozzle cover may be greater than the length of the air outlet tube 13 extending within the nozzle cover.
  • the peristaltic pump tube 2 of the active venting assembly can be clamped into the slot of the peristaltic pump head 6.
  • the peristaltic pump head 6 is mounted on a peristaltic pump drive.
  • a gas pressure regulator 4 may be disposed on the airway tube 3 between the gas exchanger 1 and the outlet tube 13 of the cell culture flask 28. The inner diameter of the airway tube 3 is adjusted by the gas pressure regulator 4 to adjust the air pressure in the cell culture flask 28, thereby providing a high pressure culture environment.
  • FIG 19 is a schematic view showing the structure of still another exemplary embodiment of the cell culture device of the present invention.
  • the cell culture device of this embodiment may be a multi-layer cell culture device such as a cell factory.
  • the active venting assembly of the present invention can have two interface devices 11. One of the two interface devices may be provided with an intake pipe 12, and the other of the two interface devices may be provided with an air outlet pipe 13. Intake tube 12 and outlet tube 13 are coupled to the inlet channel interface and the exhaust channel interface of the multi-layer cell culture, respectively, to form a closed gas circulation loop.
  • the multi-layer cell culture device of this embodiment can be used for large-scale cell culture and production of biotechnological products.
  • FIG 20 is a schematic view showing the state of use of the bioreactor and cell culture device according to the present invention.
  • the cell culture vessel or bioreactor is oversized, for example, larger than the volume of the incubator.
  • the volume and power of the peristaltic pump or/and the shaker are too large, causing the heat generation to affect the temperature control effect of the incubator.
  • bioreactors and cell cultures should not be placed in a humid environment in carbon dioxide and three gas incubators. In these cases, it is possible to place only the gas exchanger of the active venting assembly in a carbon dioxide incubator or a three-gas incubator, while placing the remaining devices (eg bioreactor, cell culture, peristaltic pump, shaker, etc.) Outside the incubator.
  • a device including a large bioreactor 25, a multi-layer cell culture device 29, a large peristaltic pump 17 having a plurality of pump heads, and a large-scale constant temperature shaker 31 can be placed in a carbon dioxide incubator or a three-gas culture. Outside the box.
  • multiple gas exchangers can be used in series and/or in parallel.
  • a plurality of air tubes can be branched from one air duct through a three-way joint or a multi-way joint to connect two or more gas exchangers in parallel.
  • the gas exchange surface area can be increased, thereby increasing gas exchange efficiency.
  • a peristaltic pump tube having a plurality of pump heads can be used to simultaneously drive a peristaltic pump tube on a plurality of branched air tubes.
  • two or more cell culture or bioreactor inlet and outlet tubes may be connected in series or in parallel to increase the scale of cell culture or biological response.
  • multiple bioreactors and/or cell culturers can be separately coupled to a plurality of active venting assemblies, with a peristaltic pump having multiple pump heads simultaneously achieving gas delivery. As described above, it is possible to place only the gas exchanger of the gas delivery device in a carbon dioxide incubator or a three-gas incubator.
  • the inlet and outlet tubes of the membrane bioreactor and the liquid inlet and outlet tubes may be disposed at the fusion edge of the soft membrane or at the upper planar soft membrane.
  • a nozzle cap that mates with a nozzle on the membrane bioreactor can be provided to effect communication of the reactive venting assembly with the membrane reactor.
  • a gas pressure measuring device can be provided to measure the gas pressure in the bioreactor and cell culture device with a reactive venting assembly, a bioreactor, and/or a cell culture device, and control the gas drive based on the measured gas pressure. Operation of the device. For example, if the measured gas pressure is less than a preset value, the increase in the rotational speed of the peristaltic pump can be controlled.
  • the gas pressure measuring device may be a gas pressure sensor.
  • the gas pressure sensor can be placed at one or more locations in the gas pressure regulator, line, or vessel.
  • the gas pressure measuring device can be non-contact.
  • the active venting module, bioreactor, and bioreactor can be used for the bioreactor and cell culture device after production of the bioreactor and cell culture device with a reactive venting assembly, bioreactor, and/or cell culture device.
  • the cell culture device is sterilized and aseptically packaged to ensure sterility in the bioreactor and cell culture device with the active venting module, bioreactor and/or cell culture device.
  • the sterilization may be ultraviolet disinfection, high temperature sterilization, ozone sterilization, or the like.

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Abstract

La présente invention concerne un ensemble de ventilation active pour un bioréacteur et un dispositif de culture de cellules. Après avoir été accouplé à un récipient de fermentation biologique ou de culture de cellules, l'ensemble de ventilation active peut fournir un passage d'écoulement de gaz circulant pour l'échange de gaz type ventilation active. L'invention concerne en outre un bioréacteur et un dispositif de culture de cellules, qui comprennent tous deux l'ensemble de ventilation active. L'ensemble de ventilation active et le bioréacteur ou le dispositif de culture de cellules de la présente invention peuvent être placés dans un incubateur de dioxyde de carbone ou un incubateur à trois gaz pour l'utilisation.
PCT/CN2019/084203 2018-04-26 2019-04-25 Ensemble de ventilation active, bioréacteur de type ventilation active et dispositif de culture de cellules Ceased WO2019206207A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201810386282.1 2018-04-26
CN201810386282.1A CN108315256B (zh) 2018-04-26 2018-04-26 活性通气组件及其活性通气式生物反应器和细胞培养器

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WO2019206207A1 true WO2019206207A1 (fr) 2019-10-31

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PCT/CN2019/084203 Ceased WO2019206207A1 (fr) 2018-04-26 2019-04-25 Ensemble de ventilation active, bioréacteur de type ventilation active et dispositif de culture de cellules

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CN (1) CN108315256B (fr)
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Cited By (5)

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US12536415B2 (en) 2020-02-03 2026-01-27 Stamm Vegh Corporation Platform, systems, and devices for 3D printing utilizing a static optical assembly and procedural modeling applications representing 3D scenes
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CN118909731A (zh) * 2024-08-22 2024-11-08 江苏奕农生物股份有限公司 一种生物酶培养用反应设备及方法

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