Apparatus and method for controlling a bioreactor

Abstract

The present invention provides a bioreactor system that allows for the growth of cells or tissues while measuring various parameters, and that enables the control of the behavior of various parameters during the operation of the bioreactor system. [Solution] The bioreactor system may include at least one bioreactor chamber, at least one reservoir, a plurality of sensors, and a fluid circuit. The operating methods disclosed herein relate to growing cells or tissues while measuring various parameters, and to controlling the operation of various parameters during the operation of the bioreactor system. Parameters whose operation is controlled include, for example, cell concentration, flow rate, volume, pH, temperature, oxygen level, carbon dioxide level, bicarbonate ion level, nutrient compounds, and any combination thereof.

Description

Technical Field

[0001] The present disclosure generally relates to a bioreactor system configured for one or more control schemes of a biological reaction.

Background Art

[0002] Generally, a bioreactor can be used to culture microorganisms and living cells in an enclosed and controlled environment. Generally, the culturing and processing of such microorganisms and cells may require several steps, and various steps can be performed before and / or during culturing while monitoring specific parameters.

Summary of the Invention

[0003] Growing various specific cells or tissues in a bioreactor can be difficult. The present disclosure generally relates to a bioreactor system and a control scheme for the bioreactor system.

[0004] In some embodiments, the method disclosed herein includes the steps of operating at least one reservoir, at least one bioreactor, and a plurality of pumps, the steps of growing a plurality of cells in at least one bioreactor, and measuring at least three parameters in at least one bioreactor by a plurality of sensors to obtain measurements of at least three parameters, wherein the at least three parameters are: the level of cell concentration of the plurality of cells; the flow rate of a first fluid of a first medium of at least one bioreactor into the reservoir chamber of at least one reservoir; the flow rate of a second fluid of a second medium from at least one reservoir chamber into the bioreactor chamber of at least one bioreactor; the level of at least one first gas in the first medium; the level of at least one first nutrient in the first medium; the first volume of the first fluid of the first medium; and The method includes the steps of: providing a first pH of a first fluid in a culture medium; a first temperature of a first fluid in a first culture medium; or any combination thereof; providing a setpoint for each of at least three parameters of at least one bioreactor, wherein the setpoint corresponds to a predetermined range of levels of the at least three parameters; comparing a measurement of the at least three parameters to a predetermined level of the at least three parameters; and simultaneously controlling the at least three parameters by adjusting at least a first volume of the first fluid until each measurement of the at least three parameters is equal to a predetermined level of the at least three parameters, wherein a plurality of pumps and at least one reservoir are configured to remove at least a portion of the first fluid from the bioreactor or add at least a portion of a second fluid to the first fluid so that the volume of the first fluid is adjustable.

[0005] In some embodiments disclosed herein, the bioreactor system comprises at least one bioreactor chamber, at least one reservoir, a plurality of sensors, and a fluid circuit. In some embodiments, the fluid circuit comprises a first section of the fluid circuit, which is configured to fluidly connect the bioreactor chamber to at least one reservoir and to allow a first fluid contained in the bioreactor chamber to flow into the at least one reservoir; and a second section of the fluid circuit, which is configured to fluidly connect at least one reservoir to the bioreactor chamber and to allow a second fluid contained in at least one reservoir to flow into the bioreactor chamber.

[0006] In some embodiments disclosed herein, the method disclosed herein includes the step of operating a bioreactor system. In some embodiments, the step of operating a bioreactor system includes the step of acquiring sensor readings of at least three parameters via a plurality of sensors, and the step of providing a predetermined set point for each of the at least three parameters. In some embodiments, the steps of acquiring and providing may be in any order. In some embodiments, the steps of acquiring and providing are performed in a specific order. In some embodiments, the acquisition step is performed before the providing step. In some embodiments, the providing step is performed before the acquisition step. In some embodiments, after the steps of acquiring and providing, the method further includes the steps of comparing the sensor readings to a predetermined set point for at least three parameters, and controlling a fluid circuit by removing a portion of a first fluid from the bioreactor chamber, adding a portion of a second fluid to the bioreactor chamber, or a combination thereof, until each of the sensor readings substantially matches the predetermined set point for at least three parameters.

[0007] In some embodiments disclosed herein, a bioreactor system comprises a bioreactor chamber and a plurality of reservoirs, the plurality of reservoirs comprising a first reservoir and a second reservoir. In some embodiments of the bioreactor system, the system further comprises a plurality of sensors and a fluid circuit, the fluid circuit comprising a first section of the fluid circuit, which is configured to fluidly connect the bioreactor chamber to the first reservoir and to allow a first fluid contained in the bioreactor chamber to flow into the first reservoir; and a second section of the fluid circuit, which is configured to fluidly connect a second reservoir to the bioreactor chamber and to allow a second fluid contained in the second reservoir to flow into the bioreactor chamber.

[0008] In some embodiments of the methods disclosed herein, the method includes the step of operating a bioreactor system, the step of acquiring sensor readings of at least three parameters via a plurality of sensors, the step of providing a predetermined setpoint for each of the at least three parameters, the step of comparing the sensor readings for the at least three parameters with the predetermined setpoint, and the step of controlling a fluid circuit. In some embodiments, the step of controlling the fluid circuit includes removing a portion of a first fluid from the bioreactor chamber, adding a portion of a second fluid to the bioreactor chamber, or a combination thereof, until each of the sensor readings substantially matches a predetermined setpoint for at least three parameters.

[0009] In some embodiments, at least three parameters are selected from the level of cell concentration contained in the bioreactor chamber, the flow rate of a first fluid into at least one reservoir, the flow rate of a second fluid into the bioreactor chamber, the volume of the first fluid, the pH of the first fluid, the temperature of the first fluid, the level of dissolved oxygen in the first fluid, the level of dissolved CO2 in the first fluid, the level of HCO3 in the first fluid, and the level of nutrients in the first fluid. In some embodiments, the level of nutrients includes the amount or concentration of nutrients. In some embodiments, the nutrients include at least one of glucose, lactate, glutamine, glutamate, or any combination thereof. In some embodiments, the nutrients include glucose but do not include lactate, glutamine, and glutamate. In some embodiments, the nutrients include lactate but do not include glucose, glutamine, and glutamate. In some embodiments, the nutrients include glutamine but do not include glucose, lactate, and glutamate. In some embodiments, the nutrients include glutamates but do not include glucose, lactate, and glutamine.

[0010] In some embodiments, the bioreactor system includes a plurality of cells housed in a bioreactor chamber. In some embodiments, the first fluid is a liquid, gas, nutrients, culture medium, or a combination thereof. In some embodiments, the second fluid is a liquid, gas, nutrients, culture medium, or a combination thereof.

[0011] In some embodiments, the step of controlling the fluid circuit includes the step of adjusting one to three of at least three parameters, and the fluid circuit automatically adjusts one to three of the at least three parameters.

[0012] In some embodiments, the step of controlling the fluid circuit includes adjusting all of at least three parameters, and the fluid circuit automatically adjusts all of the at least three parameters. In some embodiments, the adjustments are simultaneous.

[0013] In some embodiments, the step of operating the bioreactor system is performed for a sufficient amount of time to obtain a proliferation ratio of 1.5 to 10,000 cells. In some embodiments, the step of operating the bioreactor system is performed for a sufficient amount of time to obtain a proliferation ratio of 100 to 7,500 cells. In some embodiments, the step of operating the bioreactor system is performed for a sufficient amount of time to obtain a proliferation ratio of 500 to 2,500 cells. In some embodiments, the step of operating the bioreactor system is performed for a sufficient amount of time to obtain a proliferation ratio of 50 to 1,000 cells.

[0014] In some embodiments, the bioreactor system is configured to have at least two culture modes selected from a recirculation culture mode, a perfusion culture mode, a batch culture mode, and a fed-batch culture mode. In some embodiments of the methods disclosed herein, the step of operating the bioreactor system includes changing at least two culture modes from one to the other.

[0015] In some embodiments of the bioreactor systems disclosed herein, the system comprises at least one bioreactor chamber, at least one reservoir, a plurality of sensors, a first controlled fluid path connected to the bioreactor chamber and at least one reservoir and configured to allow fluid to flow from at least one reservoir to the bioreactor chamber, a second controlled fluid path connected to the bioreactor chamber and at least one reservoir and configured to allow fluid to flow from the bioreactor chamber to at least one reservoir, and a control device. In some embodiments, the control device is configured to communicate with the plurality of sensors and to receive a plurality of parameters from the plurality of sensors, and the control device is configured to automatically control the first controlled fluid path, the second controlled fluid path, or both based on the plurality of parameters received from the plurality of sensors.

[0016] In some embodiments of the bioreactor system, the multiple parameters include at least three selected from the following: the level of cell concentration contained in the bioreactor chamber, the flow rate of a first fluid into at least one reservoir, the flow rate of a second fluid into the bioreactor chamber, the volume of the first fluid, the pH of the first fluid, the temperature of the first fluid, the level of dissolved oxygen in the first fluid, the level of dissolved CO2 in the first fluid, the level of HCO3 in the first fluid, and the level of nutrients in the first fluid. In some embodiments, the level of nutrients includes the amount or concentration of nutrients. In some embodiments, the nutrients include at least one of glucose, lactate, glutamine, glutamate, or any combination thereof. In some embodiments, the nutrients include glucose but do not include lactate, glutamine, and glutamate. In some embodiments, the nutrients include lactate but do not include glucose, glutamine, and glutamate. In some embodiments, the nutrients include glutamine but do not include glucose, lactate, and glutamate. In some embodiments, the nutrients include glutamates but do not include glucose, lactate, and glutamine.

[0017] In some embodiments, the bioreactor system is configured to operate for a sufficient time to obtain a cell proliferation ratio of 1.5 to 10,000. In some embodiments, the bioreactor system is configured to operate for a sufficient time to obtain a cell proliferation ratio of 100 to 7,500. In some embodiments, the bioreactor system is configured to operate for a sufficient time to obtain a cell proliferation ratio of 500 to 2,500. In some embodiments, the bioreactor system is configured to operate for a sufficient time to obtain a cell proliferation ratio of 50 to 1,000.

[0018] 0018

[0019] In some embodiments, the bioreactor system is configured to have at least two culture modes selected from a recirculation culture mode, a perfusion culture mode, a batch culture mode, and a fed-batch culture mode. In some embodiments, the bioreactor system is configured to switch modes from one of the at least two culture modes to the other of the at least two culture modes. In some embodiments, the bioreactor system includes a control device configured to switch modes from one of the at least two culture modes to the other of the at least two culture modes during operation.

[0020] 0020

[0021] 0021

[0022] In some embodiments, the method is performed, and the steps are carried out over a sufficient period of time to obtain a proliferation ratio of 1.5 to 10,000 cells.

[0023] The embodiments disclosed herein are shown below and can result in a variety of improved outcomes as described herein. [Brief explanation of the drawing]

[0024] Various embodiments of this disclosure can be further described with reference to the accompanying drawings, and similar structures are referred to by similar reference numerals in some of the drawings. The drawings shown are not necessarily to scale and instead focus on illustrating the general principles of this disclosure. Accordingly, certain structural and functional details disclosed herein should not be construed as necessarily limiting, but rather as representative grounds for teaching those skilled in the art to use one or more exemplary embodiments in various ways. [Figure 1] This is a schematic block diagram showing various components of a bioreactor system according to several embodiments of the bioreactor system of the present disclosure. [Figure 2A]A graph showing the results of an exemplary embodiment of the bioreactor control system of the present disclosure, illustrating some exemplary aspects of at least some embodiments of the present disclosure. [Figure 2B] Another graph showing the results of an exemplary embodiment of the bioreactor control system of the present disclosure, illustrating some exemplary aspects of at least some embodiments of the present disclosure. [Figure 2C] Yet another graph showing the results of an exemplary embodiment of the bioreactor control system of the present disclosure, illustrating some exemplary aspects of at least some embodiments of the present disclosure. [Figure 3A] An additional graph showing the results of an exemplary embodiment of the bioreactor control system of the present disclosure, illustrating some exemplary aspects of at least some embodiments of the present disclosure. [Figure 3B] Another additional graph showing the results of an exemplary embodiment of the bioreactor control system of the present disclosure, illustrating some exemplary aspects of at least some embodiments of the present disclosure. [Figure 3C] Yet another additional graph showing the results of an exemplary embodiment of the bioreactor control system of the present disclosure, illustrating some exemplary aspects of at least some embodiments of the present disclosure. [Figure 3D] Still another additional graph showing the results of an exemplary embodiment of the bioreactor control system of the present disclosure, illustrating some exemplary aspects of at least some embodiments of the present disclosure. [Figure 4] A schematic block diagram showing various components of the bioreactor system according to some embodiments of the bioreactor system of the present disclosure. [Figure 5] Another schematic block diagram showing various components of the bioreactor system according to some embodiments of the bioreactor system of the present disclosure. [Figure 6] A graph showing the results of an exemplary embodiment of the bioreactor control system of the present disclosure, illustrating some exemplary aspects of at least some embodiments of the present disclosure. [Figure 7] This is an exemplary flowchart of one embodiment of the method disclosed herein. [Modes for carrying out the invention]

[0025] Various detailed embodiments of the present disclosure are disclosed herein in conjunction with the accompanying drawings. However, it should be understood that the disclosed embodiments are merely illustrative. In addition, each of the examples given in relation to the various embodiments of the present disclosure is intended to be illustrative and not limiting.

[0026] Throughout this specification, the following terms have the meanings expressly relating to this specification unless otherwise clearly indicated by the context. The phrases “in one embodiment” and “in some embodiments” as used herein do not necessarily refer to the same embodiment, but may do so. Furthermore, the phrases “in another embodiment” and “in some other embodiments” as used herein do not necessarily refer to different embodiments, but may do so. Therefore, various embodiments can be readily combined without departing from the scope or spirit of this disclosure, as described below.

[0027] In addition, the term “based on” is not exclusive and may be based on additional factors not explicitly stated unless otherwise indicated in the context. Furthermore, throughout this specification, the meanings of “a,” “an,” and “the” include multiple references. The meaning of “in” includes “inside” and “on.”

[0028] As used herein, the term “fluid” refers to a liquid or gas. As used herein, the term “culture medium” refers to a fluid or combination of fluids on which cells can grow. As used herein, the term “flow” or “flows” refers to a fluid that deforms continuously under applied pressure and / or applied shear stress. In addition, as used herein, “flow rate” is the rate at which a substance flows per unit of time. Also as used herein, the flow or movement of a substance entering the system (inlet), exiting the system (outlet), and into and out of the pumps between components of an exemplary bioreactor system is by any suitable one-way valve that enables the transfer of the substance. As used herein, when two chambers are “fluid-connected,” this means that a fluid can flow back and forth between the two chambers. Furthermore, the term “fluid-connected” may also mean, based on some configuration of some embodiments, that the flow of a fluid may have directionality in a particular direction (e.g., from one chamber to another).

[0029] As used herein, “cell supply” or “cell supply” refers to the introduction of material into a bioreactor, the introduced material promoting cell proliferation. As used herein, “waste medium,” “waste,” or “used waste” refers to any substance secreted by cells during cell proliferation that, if present in the cell medium, inhibits cell proliferation.

[0030] As used herein, “batch culture mode” refers to supplying a predetermined amount of culture medium to a bioreactor system and supplying cells using this fresh or new medium. Waste generated by this culture mode may be generated at the same rate as new medium is received, and the batch mode is completed when all medium has been consumed and become waste. Recirculation does not occur in batch culture mode.

[0031] As used herein, "fed-batch culture mode" is the same as batch culture mode, except that new medium is introduced after each cycle in which all fresh medium has been consumed. The cycle continues for a predetermined amount of time. Recirculation does not occur in fed-batch culture mode.

[0032] As used herein, “perfusion culture mode” refers to the simultaneous supply and removal of an equal volume of fluid culture medium from the bioreactor system while cells are retained within the bioreactor chamber. This culture mode provides a constant supply and constant removal of cell waste.

[0033] As used herein, “recirculating culture mode” refers to the continuous operation of the bioreactor, in which the culture medium is circulated only between the two chambers.

[0034] As used herein, “setpoint” or “setpoints” refers to a measurement state that the control system is attempting to achieve, and the control system modifies its parameters to satisfy the setpoint or range of setpoints.

[0035] As used herein, “level” or “levels” refers to a value or a range of values. For example, “pH level” can mean a specific pH value or a specific pH range.

[0036] In this disclosure, several embodiments relate to cell culture processing and manipulation systems, including bioreactors and bioreactor systems designed for culturing cells. In some embodiments, the exemplary bioreactor system may be configured to allow all necessary steps—selection, culture, modification, activation, growth, washing, concentration, and formulation—to be performed sequentially within a single unit. In some embodiments, the exemplary bioreactor system may be able to adjust various chemical parameters as needed for culturing cells. According to some embodiments, the exemplary bioreactor system can be used in various culture modes, including, but not limited to, batch mode, fed-batch mode, perfusion mode, recirculation mode, or any combination thereof. In some embodiments, the exemplary bioreactor system may be fully controlled in a closed, sterile environment and may be implemented for single-use (discarded after one culture cycle) and multi-cycle use.

[0037] In some embodiments, but not limited to, any product of cells in the exemplary bioreactor system can be collected, including secretory factors (e.g., exosomes, growth factors, e.g., FGF, PDGF, and cytokines, e.g., IL2, TNFalfa), proteins, peptides, antibiotics, or amino acids. In some embodiments, the exemplary bioreactor system can provide optimal and adaptive culture, allow cell manipulation to be performed in a closed system, and allow for automation of the operation. In some embodiments, the high growth density achieved in the exemplary bioreactor system may be twice that observed using standard culture conditions. In some embodiments, the high growth density achieved in the exemplary bioreactor system may be five times that observed using standard culture conditions. In some embodiments, the high growth density achieved in the exemplary bioreactor system may be more than ten times that observed using standard culture conditions.

[0038] Figure 1 schematically shows an exemplary bioreactor system 100 of the present disclosure. In some embodiments, the exemplary bioreactor system has a bioreactor 103 which may also have an outlet 105. In some embodiments, the reservoir 107 is fluidly connected to the bioreactor 103 by a fluid circuit (e.g., a pump system) comprising at least two sections (e.g., two pumps 109a, 109b). The fluid circuit of the exemplary bioreactor system comprises at least two sections configured, for example, each having a pump (109a, 109b), and the fluid circuit or pump system will be collectively referred to as 109 below. In some embodiments, the reservoir 107 may further comprise an inlet 111. In some embodiments, the reservoir 107 may be another chamber having a fluid medium. In some embodiments, the fluid medium in the reservoir may be the same as the fluid medium in the bioreactor. In some embodiments, the fluid medium in the reservoir may be different from the fluid medium in the bioreactor. In some embodiments, the reservoir 107 does not contain cells. In some embodiments, the reservoir is a container configured to contain a culture medium that provides fluid delivery to the bioreactor 103. In some embodiments, the reservoir is a container configured to receive and contain waste from the bioreactor 103. In some embodiments, the bioreactor system 100 includes at least two reservoirs, i.e., a first reservoir 107 configured to contain a culture medium that supplies fluid to the bioreactor 103, the first reservoir 107 through which the fluid is received via an inlet 111, and a second reservoir (not shown in Figure 1) configured to receive and contain waste from the bioreactor 103 via a fluid circuit (not shown in Figure 1) or via an outlet 105.

[0039] In some embodiments of the exemplary bioreactor system, the bioreactor 103 further comprises an inner chamber 103a, the inner chamber 103a configured to accommodate at least a number of cells. In some embodiments, the bioreactor 103 includes a first fluid medium. In some embodiments, the first fluid medium may include at least one gas, at least one nutrient, a liquid, or any combination thereof, wherein at least one nutrient is present in an amount sufficient to supply a number of cells. The liquid of the first fluid medium may further include the volume of liquid in the bioreactor 103. In some embodiments, the liquid may have a temperature in the range of 37°C to 42°C. In some embodiments, the liquid may have a temperature in the range of 24°C to 42°C. In some embodiments, the liquid may have a pH level in the range of 6.5 pH to 7.5 pH. In some embodiments, the liquid may have a pH level in the range of 5 pH to 8 pH.

[0040] In some embodiments of the exemplary bioreactor system, the reservoir has an inner chamber 107a configured to contain at least a second fluid medium. The second fluid medium may contain at least one gas, at least one nutrient in an amount sufficient to supply at least several cells, a liquid, or any combination thereof. The liquid of the second medium may further include the volume, temperature, and pH level of the liquid in the reservoir 107. Nutrients that can be used for culturing in the exemplary bioreactor system include, but are not limited to, glucose, lactate, glutamine, glutamate, or combinations thereof. One or more gases that can be used for culturing in the exemplary bioreactor system include, but are not limited to, oxygen, nitrogen, carbon dioxide, air, or any combination thereof. In some embodiments, one or more gases are dissolved gases (e.g., dissolved in the medium).

[0041] In some embodiments, the bioreactor 103 may further comprise at least two sensors (not shown) configured to measure multiple physical and chemical parameters in the fluid medium and cells contained within the bioreactor. In some embodiments, the bioreactor 103 may further comprise at least three sensors. In some embodiments, the bioreactor 103 may further comprise at least four sensors. In some embodiments, the bioreactor may comprise five or more sensors. In some embodiments, the reservoir 107 of the exemplary bioreactor system may include at least one sensor (not shown) configured to measure both physical and chemical parameters in the fluid medium contained within the reservoir. In some embodiments, the reservoir 107 may further comprise at least two sensors. In some embodiments, the reservoir 107 may further comprise at least three sensors. In some embodiments, the reservoir 107 may further comprise at least four sensors. In some embodiments, the reservoir 107 may further comprise five or more sensors.

[0042] In some embodiments, the parameters detected and measured (e.g., via sensors) in the exemplary bioreactor system can be selected from, but are not limited to, at least: levels of cell concentration; levels of at least one nutrient; levels of at least one gas; volume of liquid in the first medium; pH level of liquid in the first medium; temperature of liquid in the first medium; or any combination thereof.

[0043] In some embodiments, the parameters are detected, detected, measured, controlled, or any combination thereof. In some embodiments of exemplary bioreactor systems, these parameters are selected from, but are not limited to, at least: the level of cell concentration contained in the bioreactor chamber; the flow rate of fluid into the reservoir; the flow rate of the same or different fluids into the bioreactor chamber; the volume of at least one fluid; the pH of at least one fluid; the temperature of at least one fluid; the level of dissolved oxygen in at least one fluid; the level of dissolved CO2 in at least one fluid; the level of HCO3 in at least one fluid; the level of nutrients in at least one fluid; and any combination thereof.

[0044] In some embodiments, the parameters to be detected and measured may be, but are not limited to, temperature, pH level, glucose concentration, dissolved oxygen concentration, lactate concentration, glutamine concentration, glutamate concentration, dissolved carbon dioxide concentration, HCO3 ion concentration, and any combination thereof.

[0045] In some embodiments, at least three of the above parameters are detected by sensors in the bioreactor system. In some embodiments, at least three of the above parameters are measured by the bioreactor system. In some embodiments, at least three of the above parameters are controllable by the configuration of the bioreactor system (e.g., via a control device configured to control a fluid circuit). In some embodiments, the exemplary bioreactor system can control the detected and measured parameters to a predetermined setpoint or range of the measured values. In some embodiments, the exemplary bioreactor system can control 1 to 5 parameters. In some embodiments, the exemplary bioreactor can control 5 to 10 parameters. In some embodiments, the exemplary bioreactor can control at least three parameters simultaneously, substantially simultaneously, or the inputs to control multiple parameters are not simultaneous, but the activation of the fluid circuit in the bioreactor system based on the control inputs simultaneously affects changes to these parameters.

[0046] Returning to Figure 1, the fluid circuit may include a fluid circuit 109 configured such that a first pump 109a extends from reservoir 107 into bioreactor 103 and a second pump 109b extends from at least one bioreactor into at least one reservoir. In some embodiments, the inlet 111 of reservoir 107 can be used to introduce material for bioreactor culture. In some embodiments, the outlet 105 of reservoir 107 can be used to remove used waste, culture medium, or any combination thereof.

[0047] In some embodiments, this configuration of the fluid circuit (e.g., pump system) 109 in the exemplary bioreactor system allows the reservoir 107 to control all inputs to the bioreactor via a control device or components to the bioreactor. In some embodiments, the reservoir 107 can use a dedicated pump system in conjunction with the reservoir to control the parameters of the biological reaction or cells and the first fluid medium within the bioreactor. In some embodiments, the exemplary bioreactor system can measure at least one parameter within the bioreactor and control that parameter by changing the parameter in the reservoir based on the measurement. In some embodiments of the exemplary bioreactor system, the parameter measurement is performed in the bioreactor 103 and controlled by changing the parameter in the reservoir 107. In some embodiments, the exemplary bioreactor system can use the pump system 109 to change the parameter in the reservoir, thereby influencing the change in the parameter in the bioreactor 103. In some embodiments of the exemplary bioreactor system, the parameter measurement is performed in the bioreactor 103 and controlled solely by changing the parameter in the reservoir 107. For example, in some embodiments, the pH level of the first medium in the bioreactor can be controlled to a range of 6.5 to 7.5 pH by controlling the range of the fluid medium in the reservoir to a range of 5 pH to 8 pH while controlling other setpoints. In some embodiments, the following parameters introduced by the pump system 109, namely the flow rate of the fluid from the bioreactor to the reservoir, the flow rate of the fluid from the reservoir to the bioreactor, or any combination thereof, can also be measured by a sensor in either the bioreactor or the reservoir.

[0048] In some embodiments, the exemplary bioreactor system may require more culture medium and nutrients as well as a larger culture volume. In some embodiments, the exemplary bioreactor system may be configured to allow for changes in the volume of the bioreactor, allowing for the addition of additional culture medium without the need to transfer cells to a separate container. In some embodiments, the exemplary bioreactor system may be configured to adjust the volume of culture medium contained in the bioreactor using a pump system. In some embodiments, the pump system is configured to remove at least a portion of the liquid from the bioreactor, add at least a portion of the reservoir liquid to the bioreactor liquid, or any combination thereof, so that the volume of the bioreactor liquid can be adjusted.

[0049] In some embodiments, the exemplary bioreactor system may alternately switch between culture modes, as detailed below. In some embodiments, alternating between culture modes can help the bioreactor system to control multiple parameters simultaneously.

[0050] In some embodiments, the system includes a batch culture mode. In some embodiments, the exemplary bioreactor system can carry out the batch culture mode by having a reservoir 107 receive a predetermined amount of culture medium and pump the medium into the bioreactor 102a. The bioreactor can then discharge waste through an outlet 105.

[0051] In some embodiments, the system includes a fed-batch culture mode. In some embodiments, the exemplary bioreactor system can carry out the fed-batch mode by having a reservoir 107 receive a predetermined amount of culture medium and pumping this medium into the bioreactor 102a. In some embodiments, the process is then repeated for a predetermined amount of time. In some embodiments, the predetermined amount of time is 1 day to 2 months. In some embodiments, the predetermined amount of time is 2 days to 4 months.

[0052] In some embodiments, the system includes a perfusion culture mode. In some embodiments, the exemplary bioreactor system can proceed with the perfusion culture mode by having a reservoir 107 receive the culture medium through an inlet 111 and pumping this same medium into the bioreactor 102a. Sensors detect parameters to ensure that only equivalent waste is removed through the bioreactor outlet 105.

[0053] In some embodiments, the system includes a recirculation mode. In some embodiments, the exemplary bioreactor system can carry out a recirculation culture mode by having a first pump 109a continuously pump liquid culture medium from the reservoir to the bioreactor and a second pump 109b continuously pump liquid culture medium from the bioreactor to the reservoir.

[0054] Figures 2A, 2B, and 2C show cell proliferation in the same exemplary bioreactor system as a specific non-limiting example. For all specific non-limiting examples in Figures 2A, 2B, and 2C, Group 1 is treated in perfusion mode and Group 2 is treated in recirculation mode. Furthermore, Group 1 is a 4-day culture with 80 percent DO (dissolved oxygen) in bioreactor 103 and 100 percent DO in reservoir 107. The seeding of Group 1 is 4.5 × 10⁶ 7 The yield was 1.22 × 10 8 Group 2 consisted of a 4-day culture with 80 percent DO in bioreactor 103 and 100 percent DO in reservoir 107. The seeding for Group 2 was 4.5 × 10⁶. 7 The yield was 1.33 × 10 8 That was the case.

[0055] Figure 2A shows the specific culture medium consumption in milliliters for two groups. Figure 2B shows the growth rate for the two groups. Figure 2C illustrates, with a non-limiting specific example, the control of a setpoint in the first culture medium of a bioreactor with two or more operating options to maintain its setpoint. Here, the 70 percent DO setpoint in the first culture medium of the bioreactor over two hours was achieved by perfusing the bioreactor with culture medium from the reservoir using pump 109a at a predetermined flow rate in milliliters and a predetermined DO percentage level in the reservoir. Rod 201 represents a flow rate of 5.5221 milliliters per minute in a reservoir with 80 percent DO. Rod 203 represents a flow rate of 1.775 milliliters per minute at a 100 percent DO percentage. The flexibility of the exemplary bioreactor system is demonstrated by this experiment. Multiple options in the reservoir can be compared to control at least one same parameter in the bioreactor and used to design improved methods, for example, with lower shear force and more efficient culture medium use. In some embodiments, the improved method aimed to obtain high concentrations of secreted factors and / or higher cell proliferation or correct phenotype while using efficient culture media or resources. In some embodiments, by reading the correct density, parameters, or time, secreted factors such as exosomes can be collected via a waste port.

[0056] Figures 3A, 3B, 3C, and 3D show graphs illustrating various parameters that can be controlled in an exemplary bioreactor system, by specific non-limiting examples. Figure 3A compares the pH level and control of the reservoir, indicated by 301, with the pH level in the bioreactor, indicated by 303. Figure 3B compares the temperature and control of the reservoir, indicated by 305, with the temperature of the bioreactor, indicated by 307. Similarly, Figure 3C compares the DO level and control of the reservoir, indicated by 309, with the DO level of the bioreactor, indicated by 311. Figure 3D shows cell growth from seeding to harvest when controlling specific parameters, namely temperature only; oxygen, temperature, and pH levels together; and oxygen, temperature, glucose, lactate, and pH levels together, by specific non-limiting examples, in this specific non-limiting example.

[0057] Figure 4 shows a timeline data graph of yet another embodiment of an exemplary bioreactor system, where glucose and lactate levels are controlled simultaneously by increasing the volume of fluid within the bioreactor. In some embodiments, this control method can result in more efficient use of the culture medium, as shown in the graph of Figure 4.

[0058] In the alternative embodiment shown in Figure 5, glucose and / or lactate levels are not measured in chamber A. Instead, glucose and / or lactate levels are measured only at the outlet of chamber B. As a result, the medium in A will have a higher glucose level, and the medium in chamber B will have a lower level. Furthermore, the waste outlet (from B) and the fresh medium in (A) are operated based solely on the measurements of the medium coming out of B.

[0059] Figure 6 shows several timeline data graphs 600, 602, 604, and 606 from a specific experimental run using one embodiment of a bioreactor system operated according to embodiments of the control schemes and methods disclosed herein. In this example, various parameters were detected and measured: dissolved oxygen (DO) of the fluid in the bioreactor chamber, temperature of the fluid in the bioreactor chamber, pH of the fluid in the bioreactor chamber, CO2 level of the fluid in the bioreactor chamber, and flow rate of the fluid flowing into the bioreactor chamber. Over the operating timeline from which this data was collected, cells contained in the bioreactor system proliferated. The DO graph (600) shows the events that occur over time when the DO setpoint is set to 15%. The temperature graph (602) shows the change in temperature in the bioreactor chamber over time. Also, as shown in graph (604), the flow of CO2 to the reservoir decreased over time due to the increased flow from the reservoir to the reaction chamber (606) due to the reduced need for acid caused by cellular lactic acid secretion in the reaction chamber. Therefore, as evidenced by the results and data obtained and shown from these graphs, the DO and pH in the biological reaction chamber can be controlled and influenced by the flow rate, DO level, and CO2 level in the reservoir. Alternatively, it can be seen that the fluid circuit can be influenced by controlling the set values ​​of various parameters (e.g., DO) so that the measured parameters (e.g., predetermined parameters) automatically match these set values. That is, by controlling (e.g., changing) the flow rate, the DO level in the biological reaction chamber can be controlled, for example. Furthermore, it is recognized that over time, as the number of cells housed in the biological reaction chamber increases, the amount of CO2 required in the reservoir decreases due to the increase in flow rate and the increase in acid secretion.

[0060] Figure 7 shows a flowchart of one embodiment of the method disclosed herein. An embodiment of Method 700 includes a step 702 of acquiring sensor measurements for at least three parameters via a plurality of sensors. Method 700 also includes a step 704 of providing predetermined setpoints for each of the at least three parameters. In some embodiments, the acquisition step 702 and the providing step 704 may be performed in any order. In some embodiments, the acquisition step 702 is performed before the providing step 704 is performed. In some embodiments, the acquisition step 702 is performed after the providing step 704 is performed. In some embodiments, the acquisition step 702 and the providing step 704 are performed at the same time or at approximately the same time (i.e., simultaneously). Next, according to the embodiment shown in Figure 7, the method 700 includes the step 706 of comparing sensor measurements with predetermined setpoints of at least three parameters, and then the step 708 of controlling a fluid circuit, which includes removing a portion of a first fluid from the bioreactor chamber, adding a portion of a second fluid to the bioreactor chamber, or a combination thereof, until each of the sensor measurements substantially matches a predetermined setpoint of at least three parameters. In some embodiments, other actions are taken to influence changes in the operation of the bioreactor system based on the parameter setpoints and sensor measurements.

[0061] The following embodiments enumerate embodiments disclosed herein, and any part of any embodiment may be combined with any other part of any embodiment. Appearance 1. A method, The steps include operating the bioreactor system, The bioreactor system, Bioreactor chamber and At least one reservoir, Multiple sensors, Fluid circuits and Equipped with, The aforementioned fluid circuit The fluid circuit comprises a first section, The first section is configured to fluidly connect the bioreactor chamber to the at least one reservoir and to flow the first fluid contained in the bioreactor chamber to the at least one reservoir, The fluid circuit comprises a second section, The second section is configured to fluidly connect the at least one reservoir to the bioreactor chamber and to flow the second fluid contained in the at least one reservoir into the bioreactor chamber. The step of operating the bioreactor system is: The steps include obtaining sensor measurements of at least three parameters via the plurality of sensors, The steps include providing a predetermined setting value for each of the at least three parameters, A step of comparing the sensor measurement values ​​with the predetermined set values ​​for at least three of the parameters, A step of controlling the fluid circuit, Until each of the sensor measurements substantially matches the predetermined set value of the at least three parameters, A portion of the first fluid is removed from the bioreactor chamber. A portion of the second fluid is added to the bioreactor chamber, or The steps include controlling the fluid circuit to perform those combinations and Methods that include... Embodiment 2. At least three parameters are, The level of cell concentration contained in the bioreactor chamber, Flow rate of the first fluid into at least one reservoir, The flow rate of the second fluid into the bioreactor chamber, The volume of the first fluid, pH of the first fluid described above, The temperature of the first fluid, The level of dissolved oxygen in the first fluid, The level of dissolved CO2 in the first fluid, The level of HCO3 in the first fluid, and Level of nutrients in the first fluid The method according to embodiment 1, selected from the following. Appearance 3. A method, The steps include operating the bioreactor system, The bioreactor system, Bioreactor chamber and Multiple reservoirs, Multiple sensors, Fluid circuits and Equipped with, The aforementioned multiple reservoirs, First reservoir and Second reservoir and Equipped with, The aforementioned fluid circuit The fluid circuit comprises a first section, The first section is configured to fluidly connect the bioreactor chamber to the first reservoir and to flow the first fluid contained in the bioreactor chamber to the first reservoir. The fluid circuit comprises a second section, The second section is configured to fluidly connect the second reservoir to the bioreactor chamber and to flow the second fluid contained in the second reservoir into the bioreactor chamber. The aforementioned step of performing the operation is The steps include obtaining sensor measurements of at least three parameters via the plurality of sensors, The steps include providing a predetermined setting value for each of the at least three parameters, A step of comparing the sensor measurement values ​​with the predetermined set values ​​for at least three of the parameters, A step of controlling the fluid circuit, Until each of the sensor measurements substantially matches the predetermined set value of the at least three parameters, A portion of the first fluid is removed from the bioreactor chamber. A portion of the second fluid is added to the bioreactor chamber, or The steps include controlling the fluid circuit to perform those combinations and Methods that include... Embodiment 4. At least three parameters are, The level of cell concentration contained in the bioreactor chamber, Flow rate of the first fluid into at least one reservoir, The flow rate of the second fluid into the bioreactor chamber, The volume of the first fluid, pH of the first fluid described above, The temperature of the first fluid, The level of dissolved oxygen in the first fluid, The level of dissolved CO2 in the first fluid, The level of HCO3 in the first fluid, and Level of nutrients in the first fluid The method according to embodiment 3, selected from the above. Embodiment 5. The method according to any one of Embodiments 1 to 4, wherein the bioreactor system includes a plurality of cells contained in a bioreactor chamber. Embodiment 6. The method according to any one of Embodiments 1 to 5, wherein the first fluid is a liquid, a gas, a nutrient, a culture medium, or a combination thereof. Embodiment 7. The method according to any one of Embodiments 1 to 6, wherein the second fluid is a liquid, a gas, a nutrient, a culture medium, or a combination thereof. Embodiment 8. The step of controlling a fluid circuit is: The process includes a step of adjusting one to three of at least three parameters, wherein the fluid circuit automatically adjusts the one to three of the at least three parameters. The method according to any one of embodiments 1 to 7. Embodiment 9. The step of controlling a fluid circuit is: The process includes a step of adjusting all of at least three parameters, wherein the fluid circuit automatically adjusts all of the at least three parameters. The method according to any one of embodiments 1 to 7. Embodiment 10. The method according to Embodiment 9, wherein the adjustments are performed simultaneously. Embodiment 11. The method according to any one of Embodiments 1 to 10, wherein the step of operating the bioreactor system is performed for a sufficient amount of time to obtain a proliferation ratio of 1.5 to 10,000 cells. Embodiment 12. The method according to any one of Embodiments 1 to 10, wherein the step of operating the bioreactor system is performed for a sufficient amount of time to obtain a proliferation ratio of 100 to 7,500 cells. Embodiment 13. The method according to any one of Embodiments 1 to 10, wherein the step of operating the bioreactor system is performed for a sufficient amount of time to obtain a proliferation ratio of 500 to 2,500 cells. Embodiment 14. The method according to any one of Embodiments 1 to 10, wherein the step of operating the bioreactor system is performed for a sufficient amount of time to obtain a proliferation ratio of 50 to 1,000 cells. Embodiment 15. The bioreactor system Recirculation culture mode, Perfusion culture mode, Batch culture mode, and Fed batch culture mode It is configured to have at least two culture modes selected from, The step of operating the bioreactor system is: Steps to change one of the at least two culture modes from one to the other. The method according to any one of embodiments 1 to 14, further comprising: Embodiment 16. Bioreactor chamber and At least one reservoir, Multiple sensors, A first controlled fluid flow path, A second controlled fluid channel, Control device and Equipped with, The first controlled fluid flow path is connected to the bioreactor chamber and the at least one reservoir, The first controlled fluid channel is configured to flow fluid from the at least one reservoir to the bioreactor chamber, The second controlled fluid channel is connected to the bioreactor chamber and the at least one reservoir, The second controlled fluid channel is configured to flow fluid from the bioreactor chamber to the at least one reservoir, The control device is configured to communicate with the plurality of sensors and to receive a plurality of parameters from the plurality of sensors. The control device is configured to control the first controlled fluid path, the second controlled fluid path, or both, based on the plurality of parameters received from the plurality of sensors. Bioreactor system. Apparatus 17. Multiple parameters, The level of cell concentration contained in the bioreactor chamber, Flow rate of the first fluid into at least one reservoir, The flow rate of the second fluid into the bioreactor chamber, The volume of the first fluid, pH of the first fluid described above, The temperature of the first fluid, The level of dissolved oxygen in the first fluid, The level of dissolved CO2 in the first fluid, The level of HCO3 in the first fluid, and Level of nutrients in the first fluid A bioreactor system according to embodiment 16, comprising at least three selected from the following. Embodiment 18. The bioreactor system according to Embodiment 16 or 17, wherein the first fluid is a liquid, a gas, a nutrient, or a combination thereof. Embodiment 19. The bioreactor system according to Embodiment 16 or 17, wherein the second fluid is a liquid, a gas, a nutrient, or a combination thereof. Embodiment 20. A bioreactor system according to any one of embodiments 16 to 19, wherein the bioreactor system is configured to operate for a period of time sufficient to obtain a cell proliferation ratio of 1.5 to 10,000. Embodiment 21. A bioreactor system according to any one of embodiments 16 to 18, wherein the bioreactor system is configured to operate for a period of time sufficient to obtain a cell proliferation ratio of 100 to 7,500. Embodiment 22. A bioreactor system according to any one of embodiments 16 to 18, wherein the bioreactor system is configured to operate for a period of time sufficient to obtain a cell proliferation ratio of 500 to 2,500. Embodiment 23. A bioreactor system according to any one of embodiments 16 to 18, wherein the bioreactor system is configured to operate for a period of time sufficient to obtain a cell proliferation ratio of 50 to 1,000. Appearance 24. The bioreactor system Recirculation culture mode, Perfusion culture mode, Batch culture mode, and Fed batch culture mode A bioreactor system according to any one of embodiments 16 to 23, configured to have at least two culture modes selected from the above. Embodiment 25. The bioreactor system according to Embodiment 24, wherein the control device is configured to change the mode from one of at least two culture modes to the other of at least two culture modes. Embodiment 26. The bioreactor system according to Embodiment 24, wherein the control device is configured to change the mode from one of at least two culture modes to the other of at least two culture modes during operation. Appearance 27. A method, This includes the step of obtaining a bioreactor, The bioreactor Bioreactor chamber and The bioreactor chamber comprises at least, Multiple cells, and First culture medium It is configured to accommodate, The first culture medium described above, The first liquid, The first liquid has a first volume, a first temperature, and a first pH. The first gas, and The first nutrient Includes one or more of the following: Exit and Multiple sensors and Equipped with, This includes the step of acquiring a reservoir, The aforementioned reservoir, The entrance and Reservoir chamber and Equipped with, The reservoir chamber is configured to contain a second culture medium. The second culture medium described above, The second liquid, The second liquid has a second volume, a second temperature, and a second pH. The second gas, and Second nutrient Includes one or more of the following: The step includes acquiring multiple pumps, The aforementioned multiple pumps A first pump, which extends from the reservoir to the bioreactor chamber, A second pump, which extends from the bioreactor to the reservoir, and Equipped with, This includes the step of operating the reservoir, the bioreactor, and the plurality of pumps, The aforementioned step of performing the operation is The steps include: growing the plurality of cells in the bioreactor, A step of obtaining measured values ​​of at least three parameters via the plurality of sensors, wherein the at least three parameters are The cell concentration levels of the aforementioned plurality of cells, The flow rate of the first liquid into the reservoir chamber of the reservoir, The flow rate of the second liquid into the bioreactor chamber, The amount of the first gas mentioned above, The amount of the above 1 nutrient, The first volume, The first pH, and The first temperature The selected step and Includes, The step includes providing a set value for each of the at least three parameters of the bioreactor, The setting value corresponds to a predetermined range for each of the at least three parameters, The step includes comparing the measured value for each of the at least three parameters with the predetermined range, Until each of the measured values ​​substantially falls within the predetermined range of the at least three parameters, Removing a portion of the first liquid from the bioreactor, or Adding a portion of the second liquid to the first liquid. A method comprising the step of simultaneously controlling the at least three parameters by adjusting the first volume via a method.

[0062] Although one or more embodiments of the present disclosure have been described, these embodiments are illustrative and not limiting, and it will be understood that many modifications will be apparent to those skilled in the art, including the use of various embodiments of the methodology, system / platform, and apparatus of the present invention described herein in any combination. Furthermore, the various steps may be performed in any desired order (any desired steps may be added and / or any desired steps may be omitted).

Claims

1. The steps include operating the bioreactor system, The bioreactor system comprises the following: Bioreactor chamber, At least one reservoir, Multiple sensors, and fluid circuit However, the fluid circuit includes the following: The first section of the fluid circuit, However, the first section is configured to fluidly connect the bioreactor chamber to the at least one reservoir and to flow the first fluid contained in the bioreactor chamber to the at least one reservoir, and The second section of the fluid circuit, However, the second section is configured to fluidly connect the at least one reservoir to the bioreactor chamber and to flow the second fluid contained in the at least one reservoir into the bioreactor chamber; However, the steps for operating the bioreactor system include the following: A step of acquiring sensor measurements of at least three parameters via the aforementioned plurality of sensors, A step of providing a predetermined setting value for each of the at least three parameters, A step of comparing the sensor measurement values ​​with the predetermined set values ​​for at least three of the parameters, and The following steps are performed to control the fluid circuit: A portion of the first fluid is removed from the bioreactor chamber. Adding a portion of the second fluid to the bioreactor chamber, or Those combinations, The step is carried out until each of the sensor measurements substantially matches the predetermined set values ​​of the at least three parameters.

2. The method according to claim 1, wherein at least three parameters are selected from the following: The level of cell concentration contained in the bioreactor chamber, Flow rate of the first fluid into at least one reservoir, The flow rate of the second fluid into the bioreactor chamber, Volume of the first fluid, pH of the first fluid, The temperature of the first fluid, The dissolved oxygen level of the first fluid, Dissolved CO in the first fluid ２ level, HCO in the first fluid ３ The level, and The level of nutrients in the first fluid.

3. The steps include operating the bioreactor system, The bioreactor system comprises the following: Bioreactor chamber, Multiple reservoirs equipped with the following: The first reservoir, and Second reservoir, Multiple sensors, and fluid circuit, However, the fluid circuit comprises the following: The first section of the fluid circuit, However, the first section is configured to fluidly connect the bioreactor chamber to the first reservoir and to flow the first fluid contained in the bioreactor chamber to the first reservoir. and The second section of the fluid circuit However, the second section is configured to fluidly connect the second reservoir to the bioreactor chamber and to flow the second fluid contained in the second reservoir into the bioreactor chamber. However, the steps to perform the operation include the following: A step of acquiring sensor measurements of at least three parameters via the aforementioned plurality of sensors, A step of providing a predetermined setting value for each of the at least three parameters, A step of comparing the sensor measurement values ​​with the predetermined set values ​​for at least three of the parameters, and The following steps are performed to control the fluid circuit: To remove a portion of the first fluid from the bioreactor chamber, Adding a portion of the second fluid to the bioreactor chamber, or Those combinations, The step is carried out until each of the sensor measurements substantially matches the predetermined set value of the at least three parameters.

4. The method according to claim 3, wherein at least three parameters are selected from the following: The level of cell concentration contained in the bioreactor chamber, Flow rate of the first fluid into at least one reservoir, The flow rate of the second fluid into the bioreactor chamber, Volume of the first fluid, pH of the first fluid, The temperature of the first fluid, The dissolved oxygen level of the first fluid, Dissolved CO in the first fluid ２ level, HCO in the first fluid ３ The level, and The level of nutrients in the first fluid.

5. The method according to any one of claims 1 to 4, wherein the bioreactor system includes a plurality of cells contained in a bioreactor chamber.

6. The method according to any one of claims 1 to 5, wherein the first fluid is a liquid, a gas, a nutrient, a culture medium, or a combination thereof.

7. The method according to any one of claims 1 to 6, wherein the second fluid is a liquid, a gas, a nutrient, a culture medium, or a combination thereof.

8. The step of controlling the fluid circuit is The process includes a step of adjusting one to three of at least three parameters, wherein the fluid circuit automatically adjusts the one to three of the at least three parameters. The method according to any one of claims 1 to 7.

9. The step of controlling the fluid circuit is The process includes a step of adjusting all of at least three parameters, wherein the fluid circuit automatically adjusts all of the at least three parameters. The method according to any one of claims 1 to 7.

10. The method according to claim 9, wherein the adjustment is performed simultaneously.

11. The method according to any one of claims 1 to 10, wherein the step of operating the bioreactor system is performed for a time sufficient to obtain a proliferation ratio of 1.5 to 10,000 cells.

12. The method according to any one of claims 1 to 10, wherein the step of operating the bioreactor system is performed for a time sufficient to obtain a proliferation ratio of 100 to 7,500 cells.

13. The method according to any one of claims 1 to 10, wherein the step of operating the bioreactor system is performed for a time sufficient to obtain a proliferation ratio of 500 to 2,500 cells.

14. The method according to any one of claims 1 to 10, wherein the step of operating the bioreactor system is performed for a time sufficient to obtain a proliferation ratio of 50 to 1,000 cells.

15. The method according to any one of claims 1 to 14, However, the bioreactor system is configured to include at least two culture modes selected from the following: Recirculation culture mode, Perfusion culture mode, Batch culture mode, and Fed batch culture mode The steps for operating the bioreactor system further include: A step of changing one of the at least two culture modes from one to the other.

16. A bioreactor system equipped with the following features: Bioreactor chamber; At least one reservoir; Multiple sensors; A first controlled fluid channel, however, The first controlled fluid passage is connected to the bioreactor chamber and the at least one reservoir, and The first controlled fluid channel is configured to flow fluid from the at least one reservoir to the bioreactor chamber; A second controlled fluid channel, however, The second controlled fluid channel is connected to the bioreactor chamber and the at least one reservoir, and The second controlled fluid channel is configured to flow fluid from the bioreactor chamber to the at least one reservoir. control device The control device is configured to communicate with the plurality of sensors and to receive a plurality of parameters from the plurality of sensors. The control device is configured to control the first controlled fluid path, the second controlled fluid path, or both, based on the multiple parameters received from the multiple sensors.

17. The bioreactor system according to claim 16, wherein the plurality of parameters include at least three selected from the following: The level of cell concentration contained in the bioreactor chamber, Flow rate of the first fluid into at least one reservoir, The flow rate of the second fluid into the bioreactor chamber, Volume of the first fluid, pH of the first fluid, The temperature of the first fluid, The dissolved oxygen level of the first fluid, Dissolved CO in the first fluid ２ level, HCO in the first fluid ３ The level, and The level of nutrients in the first fluid.

18. The bioreactor system according to claim 16 or 17, wherein the first fluid is a liquid, a gas, a nutrient, or a combination thereof.

19. The bioreactor system according to claim 16 or 17, wherein the second fluid is a liquid, a gas, a nutrient, or a combination thereof.

20. The bioreactor system according to any one of claims 16 to 19, wherein the bioreactor system is configured to operate for a period of time sufficient to obtain a growth rate of 1.5 to 10,000 cells.

21. The bioreactor system according to any one of claims 16 to 18, wherein the bioreactor system is configured to operate for a time sufficient to obtain a cell proliferation ratio of 100 to 7,500.

22. The bioreactor system according to any one of claims 16 to 18, wherein the bioreactor system is configured to operate for a period of time sufficient to obtain a cell proliferation ratio of 500 to 2,500.

23. The bioreactor system according to any one of claims 16 to 18, wherein the bioreactor system is configured to operate for a period of time sufficient to obtain a growth rate of 50 to 1,000 cells.

24. A bioreactor system according to any one of claims 16 to 23, wherein the bioreactor system is configured to have at least two culture modes selected from the following: Recirculation culture mode, Perfusion culture mode, Batch culture mode, and Fed batch culture mode.

25. The bioreactor system according to claim 24, wherein the control device is configured to change the mode from one of at least two culture modes to the other of at least two culture modes.

26. The bioreactor system according to claim 24, wherein the control device is configured to change the mode from one of at least two culture modes to the other of at least two culture modes during operation.

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