Bioreactor and process control system

Abstract

The present invention relates to a bioreactor for cultivating cells, comprising a housing, a reactor vessel, at least one line for transferring gases or gas mixtures to and / or from the reactor vessel with at least one reversibly or irreversibly closable closure unit, at least one line for transferring liquid systems to and / or from the reactor vessel, having at least one reversibly or irreversibly closable closure unit, and at least one rotation module designed and configured for moving the reactor vessel about the longitudinal axis thereof. The invention also relates to a process control system comprising at least one bioreactor according to the invention.

Description

[0001] Bioreactor and process control system

[0002] Description

[0003] The present invention relates to a bioreactor and a process control system containing the bioreactor.

[0004] Bioreactors as such are known to those skilled in the art.

[0005] DE 693 32 374 T2 discloses a bioreactor for cultivating human cells, comprising a cell culture chamber with a plurality of inlet openings and a plurality of substantially equidistant outlet openings, means for perfusing a liquid cell culture medium through the cell culture chamber, a gas-permeable membrane for contacting the liquid cell culture medium with an oxygen source, and means for harvesting non-adherent cells from the cell culture chamber through the flow of the culture medium. This bioreactor is intended to allow the maintenance and proliferation of human stem cells and / or early human hematopoietic cells, including complex cultures of primary cells, in a manner that allows the harvesting of the cells produced in the bioreactor without interrupting the culture.

[0006] DE 10 2017208 758 A1 relates to a device for cultivating cells, comprising a mixing device and a reaction chamber delimited by a reactor wall, which has at least one connection for the addition of gas and / or liquid. The reaction chamber must be designed such that either its internal volume, relative to a minimum internal initial volume, can be increased by at least 500% or that a horizontal cross-sectional area of ​​the reaction chamber increases from bottom to top. Such a device is intended to avoid transferring the cells into several vessels of different sizes during cell expansion. DE 10 2018 123 553 A1 discloses a bioreactor for cultivating biological cells, comprising a container for receiving a cultivation medium with a plurality of substrate filaments arranged therein, which are designed for temporary adherent coupling of the biological cells.For this purpose, the substrate filaments must be provided with a surface layer that can be switched between an adherent state, in which the biological cells can be coupled to the surface layer, and a release state, in which the adherent coupling of the biological cells to the surface layer is reduced compared to the adherent state. The bioreactor of DE 10 2018 123 553 A1 is intended to enable the cultivation of biological cells with increased efficiency and yield and / or with a gentle detachment of cells from the cultivation substrate.

[0007] DE 10 2019 100 434 A1 relates to a bioreactor for cultivating microorganisms and cells of animal or plant origin, comprising a filter bag arranged on the inner surface of a flexible wall of the bioreactor, which is bounded on the outside by the wall of the bioreactor and on the inside by a filter bag wall, and in which a spacer is arranged between the wall of the bioreactor and the filter medium. The inner filter bag wall must be formed at least partially by a filter medium, and the spacer must have at least one through-opening through which a connecting section of the filter medium must protrude and be directly connected to the wall of the bioreactor. This is intended to provide a bioreactor suitable for perfusion that can also be used for large volumes on the order of 200 litres total volume or more.

[0008] EP 3 330 368 B1 discloses a method for cultivating cells in a cell culture chamber of a bioreactor system, comprising i) introducing a cell sample into a first cell culture chamber of a first cell module of the bioreactor system, wherein the first cell module of the first cell culture chamber has an inlet, an outlet, and a connection opening, and further comprises at least one male clamping connector with a recess and at least one female connector with a raised portion, wherein the male and female connectors are arranged on the same surface of the cell module; ii) introducing a cell culture medium that promotes migration into a second cell culture chamber of a second cell module, wherein the second cell module of the second cell culture chamber has an inlet, an outlet, and a connection opening,and wherein a membrane is positioned between the open port of the first cell module and the open port of the second cell module, and wherein the first cell module and the second cell module sealingly engage, thereby securing the membrane between the first and second modules, and wherein the recess in the at least one male compression fitting of the first module and the raised portion in the at least one female compression fitting of the second cell module are sealingly engaged using a push-and-twist motion. This method is intended to enable the co-cultivation of multiple cell types in a dynamic environment in which the different cell types can communicate biochemically and yet are physically separable.

[0009] WO 2013 / 124326 A1 relates to a bioreactor system with a bioreactor connected to a solids separator. The solids separator comprises a sterilizable plastic bag through which a flow of solids passes and, within the plastic bag in the upper region, one or more internals for withdrawing a harvested stream separated from the solids from a harvested stream collection area. In the upper segment of a central region, a separation area with a separation surface is to be provided, which is inclined at an angle of 0° to 80° to the horizontal during operation. Furthermore, in the lower segment of the central region, one or more passages or internals for flow distribution of the reactor mixture are to be provided. Finally, in the lower region, a downwardly tapered solids collection area is to be provided for collecting the solids by gravity.The plant of WO 2013 / 124326 A1 is intended to provide an efficient method for the retention and recycling of animal and plant cells in a continuous or batch process, which takes into account the sensitivity of the cells with regard to mechanical shear stress and sufficient supply of the cells with nutrients.

[0010] CA 2 566 841 C refers to a bioreactor comprising a membrane support structure and a highly porous membrane resting on the membrane support structure. The membrane has a nutrient side and a so-called gas side. Furthermore, the membrane must contain an immobilized biolayer. The membrane must be configured and designed to enable the diffusion of a nutrient solution from the nutrient side to the immobilized biolayer without external pressure. Finally, the membranes must be arranged in pairs on the membrane support structure and define an interior space. The bioreactor of CA 2 566 841 C is intended to be cost-effective and durable and to enable higher bioconversion rates than conventional systems.

[0011] CA 3 086 283 A1 discloses a bioreactor for cultivating cells, comprising a base part with a first chamber containing a stirrer and a first central column removably attached to the base part, wherein the first central column forms at least part of an outer second chamber for cultivating cells and an inner third chamber for recirculating the fluid flow from the second outer chamber to the first chamber. This aims to provide a modular bioreactor with which one or more structured fixed beds can be used to simplify the manufacturing process while achieving excellent cell cultivation results in terms of homogeneity and repeatability.

[0012] Finally, DE 42 00 446 A1 discloses a bioreactor equipped with a cell carrier assembly, which can be used to optimally produce biomaterial such as cells. For this purpose, the cell carrier assembly must be at least double-walled and consist of at least two blanks made of a flat material, with the blanks forming at least one gap or gap-like region between them.

[0013] The object of the present invention is to provide a bioreactor which is not afflicted with the disadvantages of the prior art and which, in particular, can be handled easily and safely and with which adherent cells can also be cultivated.

[0014] Accordingly, a bioreactor, in particular a single-use bioreactor, for cultivating cells, in particular adherent cells, was found, comprising a housing with a housing wall and a housing base, as well as a reactor vessel with a reactor base and an opposite reactor end and a longitudinal axis extending from the reactor base to the opposite reactor end, at least one line for the transfer of gases or gas mixtures to and / or from the reactor vessel with at least one reversibly or irreversibly closable closure unit, at least one line for the transfer of liquid systems to and / or from the reactor vessel with at least one reversibly or irreversibly closable closure unit and at least one rotation module, designed and configured for the movement of the reactor vessel about the longitudinal axis.Very particular preference is given to bioreactors according to the invention, in particular single-use bioreactors, for cultivating cells, in particular adherent cells, comprising a housing with a housing wall and a housing base and a reactor vessel with a reactor base and an opposite reactor end and a longitudinal axis extending from the reactor base to the opposite reactor end, wherein the longitudinal axis has an angle of inclination relative to the horizontal, at least one line for the transfer of gases or gas mixtures to and / or from the reactor vessel with at least one reversibly or irreversibly closable closure unit, at least one line for the transfer of liquid systems to and / or from the reactor vessel with at least one reversibly or irreversibly closable closure unit and at least one rotation module, designed and configured for the movement of the reactor vessel about the longitudinal axis.

[0015] In a particularly expedient embodiment, the housing contains, in particular, the reactor vessel, the at least one line for the transfer of gases or gas mixtures to and / or from the reactor vessel, the at least one line for the transfer of liquid systems to and / or from the reactor vessel, and the at least one rotation module or a component thereof. In a particularly expedient embodiment, the housing contains, in particular, the reactor vessel, at least in sections, the at least one line for the transfer of gases or gas mixtures to the reactor vessel, at least in sections, the at least one line for the transfer of gases or gas mixtures from the reactor vessel, at least in sections, the at least one line for the transfer of liquid systems to the reactor vessel, at least in sections, the at least one line for the transfer of liquid systems from the reactor vessel, and the at least one rotation module or a component thereof.In a further particularly suitable embodiment, the housing contains in particular the reactor vessel, the at least one line for the transfer of gases or gas mixtures from the reactor vessel, the at least one line for the transfer of liquid systems from the reactor vessel and the at least one rotation module or a component thereof. Bioreactors according to the invention have also proven particularly useful in which the agitation module, in particular the vibration module, or components thereof are also present in the housing. Alternatively, and in particular additionally, it can be provided that the first pump unit or the first pump element and / or, in particular, the second pump unit or the second pump element are also present in the housing.

[0016] In a preferred embodiment, bioreactors according to the invention are also characterized in that the first pump element, in particular a pump head, preferably of a squeeze, diaphragm, or pneumatic pump, and / or, in particular, the second pump element, in particular a pump head, preferably of a squeeze, diaphragm, or pneumatic pump, are located in the housing wall. Embodiments in which at least one valve or at least one diaphragm is also present in the housing or in the housing wall have also proven particularly practical.

[0017] The closure unit of the line for the transfer of gases or gas mixtures is preferably located in or on the housing wall, or particularly preferably outside the housing. The closure unit of the line for the transfer of liquid systems to and / or from the reactor vessel can also be located outside the housing, but is preferably mounted in or on the housing wall.

[0018] In a particularly expedient embodiment, the housing can also have a housing cover in addition to the housing wall and the housing base. An embodiment without a housing cover is also possible. In one embodiment, the housing cover can be reversibly removable, particularly if the bioreactor according to the invention is to be used multiple times. Furthermore, in a preferred embodiment, the housing cover can be firmly connected to the housing wall, e.g., an integral component, and in particular form a hermetically sealed housing. This embodiment has proven particularly suitable for single-use bioreactors according to the invention.

[0019] In expedient embodiments of the bioreactor according to the invention, in addition to the at least one line for the transfer of gases or gas mixtures with the at least one closure unit to the reactor vessel, there is also at least one line for the transfer of gases or gas mixtures with the at least one closure unit from the reactor vessel. The at least one line for the transfer of gases or gas mixtures to the reactor vessel preferably represents an air, oxygen, nitrogen and / or a carbon dioxide supply line. For example, in one embodiment, an oxygen line to the reactor vessel can be provided alongside a further line for the transfer of, for example, nitrogen. Of course, any gas mixtures can also be transferred to the reactor vessel via a line.

[0020] In a further, particularly expedient embodiment of the bioreactor according to the invention, in addition to the at least one line for the transfer of liquid systems with the at least one closure unit to the reactor vessel, there is also at least one line for the transfer of liquid systems with the at least one closure unit from the reactor vessel. In this case, this at least one line for the transfer of liquid systems from the reactor vessel, preferably when designed as a bottle and particularly preferably as a cell culture bottle, represents a discharge line for cultured cells. In many cases, it has proven very advantageous to provide or integrate at least one sterile filter in the lines for the transfer of gases or gas mixtures. In a further embodiment, the lines for the transfer of liquid systems can also be equipped with a sterile filter.

[0021] The housing of the bioreactor according to the invention is preferably designed to be non-openable and / or, in particular, gas- and liquid-tight. Particularly preferably, the housing is designed to be non-openable and gas- and liquid-tight.

[0022] In the manner described above, any contamination of the environment can be excluded with the highest degree of safety. This also allows for particularly reliable handling of the bioreactor according to the invention as a module or disposable cassette, which can be easily and safely integrated into existing process control systems or processing plants. To reduce excess pressure in these bioreactors, in a preferred embodiment, the housing, in particular the housing wall, can be equipped with at least one sterile filter.

[0023] While the rotation module is primarily intended to rotate the reactor vessel around its longitudinal axis during the cultivation process, in a preferred embodiment, an agitation module, which is located at least partially within the housing and is intended to shake the reactor vessel, is connected or connectable, in particular reversibly, to a drive unit located outside the housing. In an alternative embodiment, the agitation module is located outside the housing of the bioreactor. In this case, the entire housing, including the reactor vessel, is subjected to agitation. The agitation module is intended, in particular, to initiate or facilitate the process of harvesting the cultured cells.

[0024] In a particularly suitable embodiment, the agitation module represents or comprises a vibration module. In a suitable embodiment, the agitation module and, in particular, the vibration module can represent or comprise an eccentric disc.

[0025] In a particularly advantageous embodiment, the bioreactor according to the invention comprises at least one first drive unit located in the housing or preferably outside the housing, designed and configured to drive the rotation module. Alternatively, and in particular additionally, this bioreactor according to the invention has at least one second drive unit located in the housing or preferably outside the housing, designed and configured to drive the agitation module.

[0026] In a further preferred embodiment of the bioreactor according to the invention, it is provided that the at least one line for the transfer of gases or gas mixtures to and / or from the reactor vessel, with at least one reversibly or irreversibly closable closure unit, opens into a first sterile connector present in, on, or adjacent to the housing wall. Alternatively, and in particular additionally, it can be provided that the at least one line for the transfer of gases or gas mixtures to and / or from the reactor vessel, with at least one reversibly closable closure unit, in particular adjacent to the housing wall, is equipped with a sterile filter.Furthermore, it has proven advantageous for preferred embodiments of the bioreactor according to the invention that the at least one line for the transfer of liquid systems to and / or from the reactor vessel with at least one reversibly or irreversibly closable closure unit opens into a second sterile connector present in, on or adjacent to the housing wall.

[0027] In a particularly preferred embodiment, the bioreactor according to the invention further comprises at least one internal or external, in particular flow-, liquid-, or gas-based, temperature control unit. For many applications, it has proven very expedient to integrate the at least one temperature control unit into the at least one rotation unit, in particular into at least one roller. This results in a very compact bioreactor according to the invention. In an alternative embodiment, it can also be provided to design the reactor vessel with double walls and to achieve temperature control by introducing heated liquids through the cavity thus formed.Furthermore, in particularly expedient embodiments of the bioreactor according to the invention, it can be provided that it further comprises, in particular present in the housing, at least one oxygen sensor and / or at least one carbon dioxide sensor and / or at least one lactate sensor and / or at least one glucose sensor and / or at least one fructose sensor and / or at least one temperature sensor and / or at least one pH sensor and / or at least one impedance sensor and / or at least one permittivity sensor and / or at least one capacitance sensor and / or at least one contact sensor, set up and designed to determine the opening status of the at least one closure unit.

[0028] In a suitable embodiment of the bioreactor according to the invention, one or more sensors can be located within a measuring cell, in particular a flow-through measuring cell, or can be in contact with or brought into contact with a measuring cell. This measuring cell is preferably located within the housing. The measuring cell makes it possible to withdraw fluid from the reactor vessel in order to return it to the reactor vessel after the measurement has been performed.

[0029] In the bioreactor according to the invention, at least one temperature, oxygen, pH, glucose, and / or, in particular, lactate sensor is preferably used. Measuring cells for the bioreactor according to the invention are particularly preferred which are equipped with an oxygen, pH, glucose, and / or, in particular, lactate sensor. These are preferably located in the housing.

[0030] The bioreactor according to the invention can further be equipped with at least one first pump unit or a first pump element in operative connection with the at least one line for the transfer of gases or gas mixtures to and / or from the reactor vessel and alternatively and in particular additionally with at least one second pump unit or a second pump element in operative connection with the at least one line for the transfer of liquid systems to and / or from the reactor vessel. The second pump unit is preferably designed and configured for the removal and / or addition of process media and / or reagents and / or cultured cells. The first pump element in operative connection with the at least one line for the transfer of gases or gas mixtures to and / or from the reactor vessel is preferably located in or on the housing wall.Furthermore, the second pump element can also be operatively connected to the at least one line for transferring liquid systems to and / or from the reactor vessel in or on the housing wall. With the aforementioned embodiments, a particularly user-friendly and reliably liquid- and gas-tight bioreactor according to the invention can be obtained. A pump unit can be, for example, a so-called peristaltic pump or a peristaltic squeeze pump or a diaphragm pump. Pneumatic pumps are also particularly suitable. Pneumatic pumps can be used as overpressure or vacuum pumps. The pump element of a peristaltic pump can comprise a pump head containing sliding shoes or rollers on a rotor. Such pump elements can be integrated into a housing wall orcan be located within the housing, with the pump drive being located outside the housing and being able to be separated. The previously described design of a pump element is particularly suitable for the transfer of liquids.

[0031] The bioreactor according to the invention preferably has at least one second pump unit or a second pump element operatively connected to the at least one line for transferring liquid systems to and / or from the reactor vessel. In this embodiment, the at least one first pump unit or the first pump element for transferring gases or gas mixtures to and / or from the reactor vessel does not have to be present as a component of the bioreactor at the same time. With a second pump unit or with the aid of a second pump element, liquid can be pumped both into and out of the reactor vessel. Two separate second pump units can also be used for filling and removal. The second pump unit is accordingly preferably designed and configured for the removal and / or addition of process media and / or reagents and / or cultured cells.

[0032] A particularly high degree of handling safety and ease of use can also be ensured by the bioreactor according to the invention further comprising a height adjustment unit located inside or outside, in particular at least partially inside, the housing for adjusting the angle of inclination of the longitudinal axis of the reactor vessel. In very expedient embodiments, the height adjustment unit represents or comprises an adjusting screw or an adjusting motor. By adjusting the angle of inclination, for example, the filling level of the reactor vessel of the bioreactor according to the invention can be influenced.

[0033] The reactor vessel of the bioreactor according to the invention preferably represents or comprises a bottle, in particular a cell culture bottle, with a bottle bottom and an opposite bottle end. In expedient embodiments, the cell culture bottle comprises a bottom region, a lid region, and a cylindrical region arranged therebetween. Preferably, an Archimedean screw with a tube through which liquid can flow, also called a central tube, arranged along its central axis is provided inside the cylindrical region. Preferably, there is at least one wall arranged concentrically around the tube, which extends over the cylindrical region. Particularly preferably, the cell culture bottles are formed as a single piece.Furthermore, the central tube is particularly preferably designed in such a way that it has a first opening on the outside of the cell culture flask in the lid region and a second opening inside the cell culture flask on the underside of the cylinder region. Furthermore, cell culture flasks in which the Archimedean screw is integrally connected to the cylinder region and the at least one wall are also preferred, wherein the Archimedean screw and the at least one wall are arranged such that, upon rotation of the cell culture flask, a fluid flows along the Archimedean screw from the underside of the cylinder region into the lid region.

[0034] For the rotation module of the bioreactor according to the invention, a roller module can be used expediently, comprising at least two rollers, designed and configured for receiving and rotating the reactor vessel, in particular the bottle, about its longitudinal axis, wherein the rotation module, in particular the at least two rollers, is or can be inclined at an angle of inclination relative to the horizontal.

[0035] In a particularly reliable embodiment, the roller module comprises four rollers, two rollers in pairs being designed and arranged to support the reactor vessel, in particular the bottle, and two rollers in pairs being arranged above the reactor vessel, in particular the bottle.

[0036] It can also be provided that the rotation module comprises a shaft that can be brought into engagement with the reactor vessel.

[0037] Furthermore, embodiments of the bioreactor according to the invention have proven particularly useful in which the agitation module, in particular vibration module, is integrated in the rotation module, in particular in the at least one roller, preferably in at least one pair of rollers of the roller module.

[0038] The object underlying the invention is further achieved by a process control system comprising at least one bioreactor according to the invention, in particular in the form of a disposable cassette. In this case, it can preferably be provided that the process control system according to the invention is equipped with at least one incubator unit and / or, in particular, at least one pump system.

[0039] In a preferred embodiment, the process control system according to the invention further comprises a first connection to the at least one line for the transfer of gases or gas mixtures to the reactor vessel with at least one reversibly or irreversibly closable closure unit; a second connection to the at least one line for the transfer of gases or gas mixtures from the reactor vessel with at least one reversibly or irreversibly closable closure unit; a third connection to the at least one line for the transfer of liquid systems to the reactor vessel with at least one reversibly or irreversibly closable closure unit and / or a fourth connection to the at least one line for the transfer of liquid systems from the reactor vessel with at least one reversibly or irreversibly closable closure unit.In particular, the process control system according to the invention comprises a third connection to the at least one line for the transfer of liquid systems to the reactor vessel with at least one reversibly or irreversibly closable closure unit and a fourth connection to the at least one line for the transfer of liquid systems from the reactor vessel with at least one reversibly or irreversibly closable closure unit.

[0040] In a particularly expedient embodiment, the process control system according to the invention further comprises at least one sensor, configured and designed to determine the connection and / or working status of the first, second, third or fourth connection with the respective corresponding closure element.

[0041] The bioreactor according to the invention and the process control system according to the invention readily enable GMP-compliant analysis, for example, with regard to determining the content of glucose, fructose, lactate, oxygen, and carbon dioxide. Software-controlled control of all process parameters such as pH, temperature, and humidity is also possible. Another particular advantage is that the bioreactor according to the invention, in the form of a disposable cassette, can be integrated into a process control system containing an incubator or a pump system without further cleaning or installation effort. Furthermore, based on the evaluation of the parameters analytically determined during operation of the bioreactor, the further course of the processes taking place in the reactor vessel can be fine-tuned or controlled, for example, through the targeted addition of reaction components such as glucose.It has also been found to be particularly surprising that homogeneous cell suspensions can be obtained reliably and repeatably with the bioreactors according to the invention and that cell clumping can be avoided when harvesting the generated cell material.

[0042] Another particular advantage is that leaks can be excluded and that all materials used in the bioreactor according to the invention can be made chemically inert and can also be easily sterilized.

[0043] The bioreactor according to the invention ensures that the risk of introducing, transmitting, or spreading communicable diseases during the production of propagated cells (e.g., HCT / Ps such as mesenchymal stem cells, induced pluripotent stem cells, primary cells, and cell lines) is kept extremely low or can be completely eliminated. Another advantage is that the bioreactor according to the invention allows homogeneous cell suspensions to be obtained in a fully or almost fully automated process.

[0044] Further features and advantages of the invention will become apparent from the following description, in which preferred embodiments of the invention are explained by way of example with reference to schematic drawings. In the drawings:

[0045] Figure 1 Components of the bioreactor according to the invention in perspective view;

[0046] Figure 2 is a cross-sectional view of a bioreactor according to the invention;

[0047] Figure 3 Components of a process control system according to the invention in a schematic representation; and

[0048] Figure 4 is a cross-sectional view of components of a bioreactor according to the invention.

[0049] Figure 1 shows components of the bioreactor (1) according to the invention in a perspective view. The reactor vessel (2) has a reactor base (4) and an opposite reactor end (6). The reactor vessel (2), or rather the longitudinal axis extending from the reactor base (4) to the opposite reactor end (6), is inclined relative to the horizontal when used as shown in the illustrated embodiment, so that the reactor end (6) opposite the reactor base (4) is further away from the horizontal. A reactor inlet (8) is located at the reactor end (6) opposite the reactor base (4). In the illustrated embodiment, the reactor vessel (2) has the shape of a cell culture flask. The side wall (10) of this reactor vessel (2) is held by four rotatable rollers (14) located in a frame (12). The bottle-shaped reactor vessel (2) rests on the lower pair of rollers (14).The rollers (14), like the frame (12), are part of the rotation module (15) of the bioreactor according to the invention. The rotation of the reactor vessel (2) is brought about by a drive unit (16) which, in the illustrated embodiment, is connected to the reactor base (4) via a coupling joint (not shown; see Figure 2) in the housing wall and is shown in part. Furthermore, the bioreactor according to the invention has an agitation unit or module (18), which in the illustrated embodiment includes an eccentric disc. This agitation unit (18) is connected or connectable to a drive (not shown) and is used to mechanically shake the reactor vessel (2) at periodic intervals, for example, to initiate or force the harvesting of cells cultivated in the reactor vessel (2). The degree of inclination of the reactor vessel (2) can be adjusted by a height adjustment unit (20).

[0050] Figure 2 shows a cross-sectional view through a device which essentially corresponds to the device according to Figure 1 (in Figure 1, for example, the housing 24 is not shown). In the embodiment shown, the shaft (22) from the drive unit (16) for the rotation module (15) to the connection to the bottom (4) of the reactor vessel (2) opens into an articulated coupling (25) present in the housing wall of the housing (24) of the bioreactor according to the invention. The drive unit (16) can be decoupled from the rotation module (15) via this articulated coupling. Figure 2 also shows a spring suspension (26) in connection with the bottom plate (28) of the housing (24) of the reactor vessel (2). In this way, in interaction with the movement of the eccentric disc, an even stronger vibration of the reactor vessel (2) is induced. In Figure 2, the side walls (32) and the upper cover (34) of the housing wall, iethe housing cover of the housing (24) is indicated by dashed lines. The housing cover (34) can be removable or a fixed, i.e. non-removable, part of the housing. In the embodiment shown, it is a non-removable housing cover, which can in particular be an integral part of the housing. The bioreactor according to Figure 2 can accordingly also be used as a single-use bioreactor. At the reactor inlet (30) of the reactor vessel (2) there is a line (36) for the transfer of liquid systems, which extends beyond the housing wall (32) and there opens into a closure unit (38) in the form of a sterile connector or comprises a sterile connector. Furthermore, in the embodiment shown, a line (37) for the transfer of gases or gas mixtures extends from the reactor inlet (30) to the housing wall (32), which line opens into or merges into a closure unit (40) in the form of a sterile filter.In this way, a non-openable housing (24) can be used in the bioreactor according to the invention, which is also gas- and liquid-tight.

[0051] Figure 3 shows components of an embodiment of the process control system (100) according to the invention in a schematic representation. The components of the bioreactor (1) according to the invention present in the housing (24) are not explicitly shown. The drive unit (16) of the rotation module is attached to the housing (24) or to the rear housing wall (32). At the front, a sterile connector (40) is integrated into the housing wall (32), to which a line (42) for the transfer of liquid systems is connected. A sterile filter (40) is also provided in the front housing wall (32) or - in the embodiment shown - at the end of an external line extending from the housing wall or at the end of a line passing through the housing wall. A line (44) for the transfer of gases or gas mixtures is connected to this sterile filter. The gases or gas mixtures flowing into the housing or the rear housing wall via this transfer line (44)The gases and gas mixtures that can be introduced into the reactor vessel can be heated via a temperature control unit (46) connected to this transfer line before being introduced into the reactor vessel. A mixing unit (48), which is also present in the illustrated embodiment, can be used to homogeneously mix two or more gases (A, B, C) before being introduced into the housing (24) or the reactor vessel (2).

[0052] A pump unit (50) is provided in the transfer line (42) for liquid systems, which is connected to the sterile connector (38) outside the housing. This pump unit is designed both to transfer aqueous systems such as reactor medium (52), washing solution (54), and harvesting reagent (56) from separate containers into the reactor vessel (2), and to transfer the cells cultivated in the reactor vessel (2) to a corresponding harvesting container (58) after the end of the cultivation period. Furthermore, the process control system (100) according to the invention allows the reactor vessel (2) to be cleaned with a washing solution from a separate container (54) after the cultivated cells have been harvested, and to collect the resulting liquids in a separate container (60) for proper disposal.After the cultured cells have been isolated and the reactor vessel (2) has been cleaned, the bioreactor can be disconnected from the process control system (100) and sent for separate disposal or recycling. Figure 4 shows a cross-sectional view of components of a bioreactor according to the invention. The reactor vessel (2) in the form of a cell culture flask is located in the housing (32). The rotation and agitation modules and their components are not shown in Figure 4 for the sake of clarity. A line (36) for the transfer of liquid systems leads through the reactor inlet (30) of the reactor vessel (2). This line extends beyond the housing wall (32) and there merges into a closure unit (38) containing a sterile connector. The line (36) extends into the middle part of the reactor vessel (2).In this way, it can be ensured that this line is always immersed in the culture medium during use of the bioreactor. The line (37) for the transfer of gases or gas mixtures opens into a closure unit (40) located in the housing wall (32) and containing a sterile filter (40). This line extends only slightly beyond the reactor inlet (30). This ensures that this line does not immerse itself in the culture medium during use of the bioreactor. The embodiment of a bioreactor according to the invention shown in Figure 4 also includes a measuring cell (62) located within the housing (24). Culture medium is fed to this measuring cell from the reactor vessel (2) via the liquid line 70 with the aid of a pump unit (74). After the sample material has been measured, it is returned to the reactor vessel (2) via the line (72).The measuring cell is advantageously a so-called flow-through cell. The pump unit (74) can be designed, for example, as a peristaltic pump, wherein the pump drive is located outside the housing and is operatively connected to the pump elements of the peristaltic pump by a drive shaft. The measuring cell (62) is expediently located on or in the region of the inner wall of the housing (24). In the illustrated embodiment, the measuring cell (62) has an oxygen sensor (68), a pH sensor (66), and a glucose sensor (64). These sensors are designed and configured to transmit the detected parameters, for example, to the process control system (100). In this way, adjustments to the reaction parameters can be made during the cultivation process in order to achieve an optimized yield.

[0053] The features of the invention disclosed in the above description, in the claims and in the drawings may be essential for the realization of the invention in its various embodiments, both individually and in any combination.

Claims

An sprü che 1. Bioreactor (1), in particular a single-use bioreactor, for cultivating cells, in particular adherent cells, comprising a housing (24) with a housing wall (32) and a housing base (28), as well as a reactor vessel (2) with a reactor base (4) and an opposite reactor end (6), as well as a longitudinal axis extending from the reactor base (4) to the opposite reactor end (6), wherein the longitudinal axis preferably has an angle of inclination relative to the horizontal, at least one line (37) for the transfer of gases or gas mixtures to and / or from the reactor vessel (2) with at least one reversibly or irreversibly closable closure unit (40), at least one line (36) for the transfer of liquid systems to and / or from the reactor vessel (2) with at least one reversibly or irreversibly closable closure unit (38), and at least one rotation module (15),designed and arranged for the movement of the reactor vessel (2) about the longitudinal axis., 2. Bioreactor (1) according to claim 1, further comprising at least one agitation module (18), in particular present in the housing (24), designed and configured for shaking the reactor vessel (2), wherein the agitation module (18) preferably represents or comprises a vibration module and particularly preferably represents or comprises an eccentric disc.

3. Bioreactor (1) according to claim 1 or 2, further comprising at least one first drive unit (16) present in the housing (24) or preferably outside the housing (24), designed and configured to drive the rotation module (15) and / or, in particular and, at least one second drive unit present in the housing (24) or preferably outside the housing, designed and configured to drive the agitation module (18).

4. Bioreactor (1) according to one of the preceding claims, characterized in that the at least one line (37) for the transfer of gases or gas mixtures to and / or from the reactor vessel (2) with the at least one closure unit (40) opens into a first sterile connector located in or adjacent to the housing wall and / or, in particular or, that the at least one line (37) for the transfer of gases or gas mixtures to and / or from the reactor vessel (2) with the at least one closure unit (40), in particular adjacent to the housing wall, is equipped with a sterile filter and / or, in particular and, that the at least one line (36) for the transfer of liquid systems to and / or from the reactor vessel (2) with the at least one closure unit (38) opens into a second sterile connector located in or adjacent to the housing wall.

5. Bioreactor (1) according to one of the preceding claims, further comprising at least one internal or external, in particular flow-, liquid- or gas-based, temperature control unit (46).

6. Bioreactor (1) according to one of the preceding claims, further comprising, in particular present in the housing (24) or in the reactor vessel (2), at least one oxygen sensor and / or at least one carbon dioxide sensor and / or at least one lactate sensor and / or at least one glucose sensor and / or at least one fructose sensor and / or at least one temperature sensor and / or at least one pH sensor and / or at least one impedance sensor and / or at least one permittivity sensor and / or at least one capacitance sensor and / or at least one contact sensor arranged and designed to determine the opening status of the at least one closure unit (38, 40).

7. Bioreactor (1) according to one of the preceding claims, further comprising at least one first pump unit or a first pump element in operative connection with the at least one line (37) for the transfer of gases or gas mixtures to and / or from the reactor vessel (2) and / or at least one second pump unit or a second pump element (50) in operative connection with the at least one line (36) for the transfer of liquid systems to and / or from the reactor vessel (2), wherein the at least one second pump unit is preferably designed and configured for the removal and / or addition of process media and / or reagents and / or cultured cells.

8. Bioreactor (1) according to one of the preceding claims, further comprising a height adjustment unit (20) present inside or outside the housing (24) for adjusting the angle of inclination of the longitudinal axis of the reactor vessel (2), wherein the at least one pump unit is preferably designed and configured for the removal and / or addition of process media and / or reagents and / or cultured cells.

9. Bioreactor (1) according to one of the preceding claims, characterized in that the rotation module (15) comprises or represents a roller module, comprising at least two rollers (14), designed and configured for receiving and rotating the reactor vessel (2), in particular the bottle, about its longitudinal axis, wherein the rotation module (15), in particular the at least two rollers (14), are inclined or can be inclined at an angle to the horizontal, or that the rotation module (15) comprises a shaft that can be brought into engagement with the reactor vessel (2), wherein the roller module preferably comprises four rollers (14), wherein two rollers (14) are designed and configured in pairs to support the reactor vessel (2), in particular the bottle, and that two rollers (14) are arranged in pairs above the reactor vessel (2), in particular the bottle.

10. Bioreactor (1) according to one of the preceding claims, characterized in that the agitation module (18) is or can be connected, in particular reversibly, to the housing (24).

11. Bioreactor (1) according to one of the preceding claims, characterized in that the at least one line (37) for the transfer of gases or gas mixtures to and / or from the reactor vessel (2) represents an air, oxygen, nitrogen and / or carbon dioxide supply line to the reactor vessel (2).

12. Bioreactor (1) according to one of the preceding claims, characterized in that the at least one line (36) for the transfer of liquid systems from the reactor vessel (2), in particular from the bottle, represents or comprises a discharge line for cultured cells.

13. Bioreactor (1) according to one of the preceding claims, characterized in that at least one sterile filter is provided or integrated in the lines (37) for the transfer of gases or gas mixtures and / or that at least one sterile connector is provided or integrated in the lines (36) for the transfer of liquid systems.

14. Bioreactor (1) according to one of the preceding claims, characterized in that the housing, in particular the housing wall (32), is equipped with at least one sterile filter, designed and arranged for the reduction of excess pressure in the housing.

15. Bioreactor (1) according to one of the preceding claims, further comprising at least one measuring cell (62), in particular a flow measuring cell, which is in fluid communication with the reactor vessel and is preferably present in the housing (24), containing at least one sensor, preferably an oxygen, carbon dioxide, lactate, glucose, fructose, pH, impedance, permittivity or capacitance sensor, particularly preferably an oxygen, carbon dioxide, lactate, glucose, fructose or pH sensor.

16. Bioreactor (1) according to one of the preceding claims, characterized in that in the housing (24) the reactor vessel (2), at least in sections the at least one line (37) for the transfer of gases or gas mixtures to and / or from the reactor vessel, at least in sections the at least a line (36) for the transfer of liquid systems to and / or from the reactor vessel and the at least one rotation module (15) or a component thereof.

17. Bioreactor (1) according to claim 16, characterized in that the agitation module (18), in particular vibration module, or components thereof are also present in the housing (24).

18. Bioreactor (1) according to claim 16 or 17, characterized in that the first pump unit or the first pump element and / or, in particular and, the second pump unit or the second pump element (50) are further present in the housing (24).

19. Bioreactor (1) according to claim 16 or 17, characterized in that the first pump element, in particular a pump head, preferably a squeeze, diaphragm or pneumatic pump, and / or, in particular and, the second pump element (50), in particular a pump head, preferably a squeeze, diaphragm or pneumatic pump, are present in the housing wall (32).

20. Bioreactor (1) according to one of claims 16 to 19, characterized in that at least one valve or at least one membrane is further present in the housing (24) or in the housing wall (32).

21. Bioreactor (1) according to one of the preceding claims, characterized in that the housing (24) further comprises a housing cover (34).

22. Bioreactor (1) according to claim 21, characterized in that the housing cover (34) is a reversibly removable housing cover.

23. Bioreactor (1), in particular single-use bioreactor, according to claim 21, characterized in that the housing cover (34) is firmly connected to the housing wall (32) and is in particular an integral part of the housing (24).

24. Process control system comprising at least one bioreactor (1) according to one of the preceding claims.

25. Process control system (100) according to claim 24, further comprising a receiving module for the bioreactor (1).

26. Process control system (100) according to claim 24 or 25, further comprising a first connection to the at least one line (37) for the transfer of Gases or gas mixtures to the reactor vessel (2) with at least the reversibly or irreversibly closable closure unit (40), a second connection to the at least one line (37) for the transfer of gases or gas mixtures from the reactor vessel (2) with at least the reversibly or irreversibly closable closure unit (40), a third connection to the at least one line (36) for the transfer of liquid systems to the reactor vessel (2) with at least one reversibly or irreversibly closable closure unit (38) and / or a fourth connection to the at least one line (36) for the transfer of liquid systems from the reactor vessel (2) with at least one reversibly or irreversibly closable closure unit (38).

27. Process control system (100) according to claim 26, further comprising at least one sensor arranged and designed to determine the connection and / or working status of the first, second, third or fourth connection with the respective corresponding closure element.

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