Bioreactors and bioreactor systems for cell and tissue growth

Abstract

The present invention relates to a bioreactor (10, 10', 100) adapted for rotation, the bioreactor comprising: a vessel (12) comprising: a first end (24) and a second end (26) defining a central axis (28) of the vessel (12) extending along a first direction, e.g. length direction, of the vessel (12) from the first end (24) to the second end (26); at least one wall (18, 18', 20) elongated along the first direction of the vessel (12); at least one media conduit (22, 22') defining a volume for receiving fresh media or spent media; an inner chamber defined by at least a portion of a space confined within the at least one wall (18, 18', 20) and comprising a fresh media chamber (14) and a spent media chamber (16); a cell culture chamber (30) in fluid communication with the at least one media conduit (22, 22') and the fresh media chamber (14) and / or the spent media chamber (16); and a movable wall (38) configured within the inner chamber to separate the fresh media chamber (14) from the spent media chamber (16) within the inner chamber.

Description

Technical Field

[0001] This invention relates to a bioreactor apparatus and system for growing cell cultures and tissues, particularly a bioreactor operating under omnidirectional standard gravity conditions (often incorrectly referred to as microgravity conditions) by continuously rotating a compartment of the bioreactor containing the cell cultures or tissues using a rotary device. More specifically, the invention relates to a container-type bioreactor in which fresh and waste growth media are held within an inner chamber and separated by a movable wall (such as a piston). The invention also includes a bioreactor system comprising the bioreactor and a drive unit for displacing the piston and rotating the bioreactor. Background Technology

[0002] Bioreactors used to grow cell cultures (whether of a single or multiple cell types) or tissues typically require operation under omnidirectional standard gravity conditions (i.e., gyratory induction conditions) because this preserves the differentiation state or in vivo (re)differentiation of many cell types in the culture or recoveries, such as functionality in cell lines. Such omnidirectional standard gravity conditions are induced, for example, by more or less continuously rotating the compartment containing the cell culture to prevent cell adhesion to the compartment walls (strictly speaking, rotation minimally increases gravity (centripetal acceleration)). Appropriate rotation in a fluid environment promotes cell adhesion to each other and exerts minimal shear forces on the culture. If desired, shear forces can be introduced for specific cell / tissue types by varying the rotation speed of the bioreactor. This causes cells to aggregate into colonies commonly named spheroids or organoids (collectively referred to as spheroids in this disclosure). Tissue sheets are also included under the general term spheroids because they will be similarly affected.

[0003] It is typically necessary to stop the rotation of a bioreactor to change the culture medium or add compounds (e.g., drugs or drug candidates) to compartments, such as incubation chambers or cell culture chambers. However, this leads to the problem that spheroids may settle to the bottom or adhere to the compartment walls during rotation cessation. This, in turn, reduces the availability of gases (e.g., oxygen) and nutrients to the spheroids, thus affecting their quality. Spheroids at the bottom of the settled spheroids are more affected than those at the top. Furthermore, different users will be more or less proficient in this operation, thus introducing more variability. Therefore, stopping rotation leads to a change from the active diffusion environment in a rotary bioreactor to static conditions with a limited diffusion rate, resulting in suboptimal growth conditions for the spheroid population (and the larger the spheroid size, the more severe the effect is observed). In particular, this can cause spheroids to lose important properties (such as having the desired phenotype). Therefore, it is desirable to be able to reduce the number of times rotation must be stopped, or ideally never stop rotating, so that the spheroids remain as homogeneous as possible. When the spheres are very homogeneous, a specific rotational speed can be found at which they are in a 'stationary orbit' relative to the surrounding culture medium and thus tumble very slightly in the culture medium solution: this reduces mechanical shear stress to an extremely low level, meaning the culture is essentially mechanically 'stress-free'.

[0004] The improved uniformity of the spheroids leads to better metabolic performance, which in turn enables, for example, more reliable in vitro predictive toxicology evaluations of drug candidates from cell cultures before conducting expensive clinical trials or similar studies, i.e., it produces a more reliable “filter” before entering clinical trials.

[0005] US 9850458 addresses this challenge by providing a lid as a fully accessible incubation chamber. However, while the removal of spheroids according to the teachings of this patent is faster than in the prior art, the uniformity of the spheroids is still somewhat compromised, and the bioreactor requires operator manipulation to remove the spheroids.

[0006] EP1966367 A1 discloses a bioreactor for incubating cell cultures, tissue biopsies, cell clusters, tissue-like structures, 'original tissues', or similar samples. The bioreactor is adapted to be rotated for use under microgravity conditions and is equipped with an incubation chamber having a small internal fluid volume, typically less than 1 ml. This small-volume bioreactor allows for the use of smaller quantities of reagents that may have limited supply or be expensive.

[0007] US 2004 / 0219659 discloses a bioreactor system comprising components for applying physiologically relevant translational and rotational strains to growing bioengineered tissue. This applies physical stress to the cells or tissue.

[0008] US 2013 / 0230907 discloses a bioreactor designed to generate hydrodynamic pressure, thereby subjecting cell culture objects to hydrodynamic pressure and thus applying physical stress to cells or tissues.

[0009] US 2017 / 0009207 discloses an apparatus including a bioreactor in which cells therein are exposed to physical stress via mechanical and electrical systems. Therefore, the apparatus is designed to apply physical stress and / or electrical stimulation to the cells.

[0010] US2016 / 0201037 discloses a bioreactor for cell growth, wherein the bioreactor device includes a piston. The bioreactor may include an in vivo body that divides the bioreactor into different chambers and promotes the growth of multi-tissue samples. The device is designed to allow cells to grow as layers (or layers) attached to the device. Thus, contact occurs between the device and the cells. Summary of the Invention

[0011] Therefore, the aim is to provide a bioreactor that achieves better uniformity of spheres, while avoiding stress on cell cultures during operation and enabling more repeatable manipulation of the bioreactor, i.e., a more easily operable bioreactor.

[0012] Another objective is to provide bioreactors that minimize contact between the walls of the bioreactor and cells or tissues.

[0013] Another objective is to provide a simple bioreactor that is easy to maintain aseptically.

[0014] A further objective is to provide a bioreactor capable of achieving stable suspension of the spheres produced therein, i.e., a essentially stationary orbit where shear forces are minimized.

[0015] A further objective is to provide bioreactors capable of providing constant or programmable culture medium replenishment.

[0016] A further objective is to provide a bioreactor capable of preloading the required culture medium into the bioreactor chamber, thereby further minimizing bioreactor manipulation.

[0017] At least one or more of these (and other purposes) are addressed, at least to some extent, by the aspects and implementations disclosed herein.

[0018] Therefore, according to a first aspect, a bioreactor suitable for rotation is provided, the bioreactor comprising:

[0019] Container, the container comprising:

[0020] A first end and a second end, defining a central axis of the container, the central axis extending along a first direction, such as a length direction, of the container from the first end to the second end; at least one wall extending along the first direction of the container; at least one culture medium conduit defining a volume for receiving fresh or waste culture medium;

[0021] The inner chamber is defined by at least a portion of a space confined within the at least one wall and includes a fresh culture medium chamber and a waste culture medium chamber;

[0022] A cell culture chamber, which is in fluid communication with the at least one culture medium conduit and the fresh culture medium chamber and / or the waste culture medium chamber; and

[0023] A movable wall is configured within the interior to separate the fresh culture medium chamber from the waste culture medium chamber within the interior.

[0024] In some implementation schemes,

[0025] - The at least one wall includes an inner wall and an outer wall, the inner wall and the outer wall defining (or creating) an annular compartment between the walls.

[0026] - The at least one culture medium conduit is the annular compartment, and

[0027] - The interior chamber is defined by at least a portion, for example, all of the space confined within the interior wall.

[0028] The inner and outer walls that define or create the annular compartment thus form a double-walled container in which fresh culture medium (growth medium) and waste culture medium are retained in the inner chamber of the container while being separated by a movable wall.

[0029] Preferably, the annular compartment defines the volume for receiving fresh culture medium.

[0030] Preferably, the culture chamber is in fluid communication with the annular compartment and the waste culture medium chamber.

[0031] In some alternative implementation schemes,

[0032] -The at least one wall is a single wall, and

[0033] - The at least one culture medium conduit is at least one conduit arranged inside or outside the single wall.

[0034] Therefore, in one embodiment of the first aspect, a bioreactor suitable for rotation is provided, the bioreactor comprising:

[0035] Container, the container comprising:

[0036] A first end and a second end, defining a central axis of the container, the central axis extending along a first direction, such as a length direction, of the container from the first end to the second end; at least one wall extending along the first direction of the container; at least one culture medium conduit defining a volume for receiving fresh culture medium;

[0037] The inner chamber is defined by at least a portion of a space confined within the at least one wall and includes a fresh culture medium chamber and a waste culture medium chamber;

[0038] A cell culture chamber, which is in fluid communication with the at least one culture medium conduit and the waste culture medium chamber; and

[0039] A movable wall is configured within the interior to separate the fresh culture medium chamber from the waste culture medium chamber within the interior.

[0040] The first end of the container is preferably flat and substantially vertical (or substantially perpendicular to, for example, a substantially horizontal axis of rotation). The second end of the container is preferably flat and substantially vertical (or substantially perpendicular to, for example, the substantially horizontal axis of rotation).

[0041] Therefore, the bioreactor acts as a perfusion bioreactor, whereby the cell cultures in the cell culture chamber are preferably retained and suspended without contacting the walls of the container, while providing a fresh source of culture medium supplying fresh nutrients and removing waste culture medium containing cell waste products. Unlike conventional perfusion bioreactors, the bioreactor disclosed herein is simple in construction and does not require piping to transport fresh culture medium from a fresh culture medium reservoir to the cell culture chamber and waste culture medium from the cell culture chamber to a waste culture medium reservoir, outlet, etc. Furthermore, compared to conventional perfusion bioreactors, the bioreactor of the first aspect can also be adapted to rotate to provide a rotary cell culture environment.

[0042] The bioreactor of the first aspect also allows the culture medium to be exchanged in various ways. For example, the culture medium can be provided at a constant rate; or the culture medium can be provided intermittently, for example, three times a day to mimic breakfast, lunch, and dinner; or the culture medium can be provided to maintain a specific component at a predetermined level, for example, by adapting a component sensor that initiates the culture medium exchange to the bioreactor or bioreactor system.

[0043] Alternatively, the fresh culture medium can be pre-equilibrated before being filled into the bioreactor.

[0044] In some implementations, gas (e.g., oxygen and carbon dioxide) exchange can occur through the plastic wall of the container (e.g., through polydimethylsiloxane (PDMS) or the like) or through a special filter installed in the wall of the container (preferably the wall of the cell culture chamber).

[0045] Alternatively, the fresh culture medium can be pre-equilibrated before being filled into the bioreactor.

[0046] Therefore, the bioreactor is simple, integrated, and capable of producing uniform spheroids. The bioreactor is also easier to maintain aseptically and operate, i.e., by being able to change or refill the cell culture only for a longer period, such as 14 days or longer, rather than the usual approximately every 48 hours. This period can be extended by filling the fresh culture chamber with a 'stock' solution in which one or more components expected to be depleted during culture are present at a higher concentration. For example, if the culture uses glucose, it is typically provided at a physiological concentration of 1 g / L. If the culture medium in the fresh culture chamber contains 4.5 g / L of glucose, the culture can be maintained for 4.5 times the length of time. Culture media containing both 1 g / L and 4.5 g / L are commercially available from several sources. For example, for metabolically highly active mature C3A spheroids, the culture medium is typically changed every 2 days (this may differ for other cell types). Considering a 10 mL cell culture chamber, this implies 7 culture medium changes over 14 days. This would require 70 mL of fresh culture medium. Therefore, the culture medium reservoir should be at least 70 mL. If the culture medium contains a high level of glucose (4.5 g / L), then 70 mL would be equivalent to a 63-day supply. However, other components in the culture medium may become limited during this period.

[0047] Therefore, in some embodiments, without considering the volume of the connecting elements, suitable internal dimensions for a 10 mL cell culture chamber would be approximately 3-4 cm in diameter and 1.5-0.8 cm in height, and for the culture medium chamber / reservoir would be approximately 3-4 cm in diameter and 10-5 cm in height. These can be scaled accordingly for containers of other diameters.

[0048] Typical flow rates can be calculated from these figures. For example, if 70 mL of culture medium must be changed every 14 days at a constant flow rate, the flow rate would be approximately 0.2 mL / hour. In other cases where the flow should mimic a daily 3' meal (each taking 30 minutes), the flow rate during each of the 3' meals would be 3.33 mL / hour.

[0049] In some embodiments of the first aspect, the cell culture chamber is separately arranged at the first end of the container and is provided with an inlet or outlet or valve. The inlet or outlet allows culture medium, such as liquid culture medium, to enter the cell culture chamber from the at least one culture medium conduit, such as the annular compartment or the at least one conduit. The outlet or valve allows culture medium, such as liquid culture medium, to enter the waste culture medium chamber from the culture chamber. The movable wall allows fresh culture medium to flow into the cell culture chamber through at least one culture medium conduit, while the orifice, and in particular the valve (if present), prevents the mixing of the two liquids, i.e., the cell culture medium and the waste culture medium.

[0050] The bioreactor described in the first aspect also makes the use of filters for separating the cell culture chamber from the fresh culture medium chamber, culture medium conduits, and waste culture medium chamber optional. Rotation of the bioreactor will often cause the spheroids to move away from the central axis, i.e., away from the outlet port of the cell culture chamber. Even a slow influx of culture medium will prevent the spheroids from entering the culture medium conduits. Therefore, the use of membranes in the bioreactor to prevent the spheroids from leaving the cell culture chamber is avoided, resulting in a much simpler and less expensive construction.

[0051] However, in a specific embodiment of the bioreactor according to the first aspect, the bioreactor includes a membrane to prevent at least a portion of the cell culture in the cell culture chamber from leaving the cell culture chamber. Preferably, the membrane(s) is disposed above or at least connected to the inlet and / or outlet orifices of the cell culture chamber. This allows components, cells, or spheroids to be retained in the cell culture chamber and also prevents components or cells (e.g., microorganisms) from entering or leaving the cell chamber. In some embodiments, such a membrane is located, for example, at an outlet or valve (see more below) for allowing culture medium (liquid culture medium) to enter the waste culture medium chamber from the culture chamber. Alternatively or additionally, such a membrane is located, for example, at an inlet or valve for allowing culture medium (liquid culture medium) to enter the cell culture chamber from the annular compartment or the culture medium conduit. As yet another alternative or further, such a membrane is located, for example, at a (e.g., circumferential) inlet port providing access to the cell culture chamber. In some embodiments, a single membrane (e.g., a membrane disc) may cover two or even all three of the above locations. In particular, the single membrane may cover an outlet or valve for allowing culture medium (liquid culture medium) to enter the waste culture medium chamber from the culture chamber and an inlet or valve for allowing culture medium (liquid culture medium) to enter the cell culture chamber from the annular compartment or the culture medium conduit.

[0052] Therefore, there may be situations where such a membrane is advantageous. These situations include culturing microorganisms or co-culturing microorganisms with spherical bodies that are too small to be significantly affected by gravity or the flow of the culture medium, or situations where they can be active, or situations where the user wishes to preferentially retain compounds inside or outside the cell growth chamber.

[0053] In another embodiment of the first aspect, the at least one culture medium conduit, such as the annular compartment or the at least one conduit, is provided with an inlet or outlet port. The inlet port allows culture medium (i.e., liquid or liquid culture medium) to enter the at least one culture medium conduit from the fresh culture medium chamber, and the outlet port allows culture medium to enter the cell culture chamber from the at least one culture medium conduit. The outlet port corresponds to the inlet port of the cell culture chamber. This facilitates the flow of fresh culture medium into the cell culture chamber. Therefore, the fresh culture medium chamber is provided with an outlet port for allowing culture medium, such as liquid culture medium, to enter the at least one culture medium conduit from the fresh culture medium chamber, wherein the outlet port of the at least one culture medium conduit corresponds to the aforementioned inlet port of the at least one culture medium conduit.

[0054] In another embodiment of the first aspect, at least a portion of the at least one wall is removable, preferably as a removable end of the at least one wall located at the first end of the container, to provide access to the cell culture chamber. This allows for a much simpler construction of the bioreactor and easier maintenance due to better access to the internal parts of the bioreactor, particularly the cell culture chamber and the cells or spheroids therein. The term "removable" means that it can be reversibly opened or removed. In a particular embodiment, the flat portion of the removable end of the at least one wall located at the first end of the container includes at least one inlet port and optionally a sensor, which is preferably mounted in the inlet port. In another particular embodiment, the sensor is mounted in its own position within the chamber wall.

[0055] In another embodiment of the first aspect, at least a portion of the at least one wall is transparent glass or plastic, configured to allow observation of the culture medium, cells, and spheroids contained therein, such as a removable end for such embodiments. This enables external monitoring and evaluation of the cell culture. In addition to being transparent, the glass or plastic is preferably biologically and chemically inert relative to the culture medium and cells. Furthermore, the plastic should preferably have a low affinity for binding, adsorbing, or absorbing any compounds (including, for example, bioactive agents or drug candidates) in the culture medium. Suitable plastics include polystyrene, polypropylene, and polyethylene.

[0056] In some implementations, the container including the chamber may be made of transparent plastic glass, while all other components are made of polypropylene.

[0057] In another embodiment of the first aspect (having an inner wall and an outer wall defining an annular compartment between the walls), the inner wall has a plurality of raised ridges extending along a first / length direction of the container and extending outward vertically, i.e., radially perpendicular to the first / length direction, until contacting the outer wall, and wherein the space between the raised ridges defines one or more sub-compartments within the annular compartment. The raised ridges are preferably arranged in parallel. The raised ridges enable better flow of the culture medium within the annular compartment because the annular compartment is divided into multiple sub-compartments, thus better approximating laminar flow conditions and thereby avoiding backmixing and turbulence that could impair the flow of the culture medium therein. The provision of the raised ridges described above enables a simpler construction and thus allows for the inexpensive manufacture of bioreactors.

[0058] In another embodiment of the first aspect, the container has a cylindrical or substantially cylindrical shape and is adapted to rotate about a rotation axis, such as a horizontal axis, by one or more associated rotating elements (e.g., a drive unit), the central axis extending along a first / length direction of the container. The rotation provides omnidirectional standard gravity conditions, meaning that spherical bodies formed in the cell culture chamber can easily suspend in a stable orbit without contacting the walls of the container. The horizontal rotation axis can be as follows: Figure 1 The central axis of 28 is defined in the middle.

[0059] In another embodiment of the first aspect, the movable wall is connected to a displacement element to axially displace the movable wall along the first / length direction of the container, preferably in a direction away from the cell culture chamber. In cases requiring flow reversal, the movable wall will axially displace along the first / length direction of the container in a direction toward the cell culture chamber. In a particular embodiment, the displacement element is a piston, and the movable wall is connected to the piston via a piston shaft, which coincides, for example or preferably, with the central axis extending along the first / length direction of the container. In another particular embodiment, a conduit is provided that extends from the cell culture chamber to at least the outside of the container through the center of the piston shaft. In yet another particular embodiment, the piston shaft is a conduit (such as a tube) extending from the cell culture chamber to at least the outside of the container. This enables easier and more direct sampling and monitoring of the cells in the culture chamber. Therefore, it allows for online sampling of the culture medium or cells in the chamber without relying on stopping the rotation of the bioreactor. In addition, not only can waste culture media be collected from the outside, but fresh culture media, compounds (such as bioactive molecules, drugs or drug candidates) or other solutions can also be introduced.

[0060] In another embodiment of the first aspect, the container rotates at a speed of 0.1-200 rpm. The optimal rotation speed of the container depends on many factors, including the age of the spheres (as they become larger with age and require higher rpm to achieve 'stationary orbit conditions'), temperature, and the viscosity of the culture medium used. Fine-tuning of the rpm may be necessary, for example, finely adjusting it to 0.1 rpm to achieve 'stationary orbit conditions'. The rotation should preferably be substantially free of vibration, tremor, and chatter.

[0061] In another embodiment of the first aspect, the container or a portion thereof is constructed of breathable plastic, or the container includes a breathable membrane for the exchange of gases such as oxygen and carbon dioxide. In a particular embodiment, the breathable membrane is arranged along the first or second end of the container. In another particular embodiment, the breathable membrane is arranged along the circumferential portion, i.e., the perimeter, of the cell culture chamber, preferably along the circumferential portion of the removable end of the outer wall located at the first end of the container. Preferably, the breathable membrane is a semi-permeable membrane.

[0062] While the container is preferably cylindrical and thus has a circular cross-sectional perimeter, as is the cell culture chamber disposed therein, the cross-sectional perimeter of the container may be non-circular, such as polygonal.

[0063] In another embodiment of the first aspect, a humidification system is provided between the culture chamber and the external atmosphere, i.e., the surrounding atmosphere outside the container. The humidification system includes a liquid reservoir preferably containing sterile water, an evaporation chamber such as an evaporation labyrinth, and a filter such as a breathable membrane, particularly a semi-permeable membrane. This facilitates gas exchange into the cell culture chamber while preventing water loss from the culture chamber. Preferably, the filter is the breathable membrane arranged along the circumferential portion, i.e., the perimeter, of the cell culture chamber. In one embodiment, the evaporation chamber, for example, of an evaporation labyrinth, and the liquid reservoir are arranged along the circumferential portion of the container in the same manner as the breathable membrane.

[0064] In another embodiment, the humidification system further includes an additional filter arranged in fluid communication with the liquid reservoir and the evaporation chamber, thereby allowing evaporation from the liquid reservoir into the evaporation chamber, such as an evaporation labyrinth. The additional filter is preferably arranged along the circumferential portion of the container and positioned between the liquid reservoir and the evaporation chamber. The additional filter is preferably highly permeable to allow evaporation from the liquid reservoir into the evaporation chamber. The additional filter can be a cartridge or a permeable filter. The latter is best suited for embodiments in which the additional filter is arranged along the circumferential portion of the container. This permeable filter will then have different characteristics from the semi-permeable membrane arranged along the circumferential portion of the cell chamber.

[0065] In one particular embodiment, the humidification system further includes at least one open port connecting the evaporation chamber, such as the evaporation labyrinth, to the outside atmosphere.

[0066] In a particular embodiment, a port is provided to allow air to be drawn into the waste culture medium chamber. This may be necessary if negative pressure occurs in the container.

[0067] In another embodiment of the first aspect, the cell culture chamber includes at least one inlet port and an optional sensor, which is preferably removably mounted in the inlet port. Mounting means integrating the sensor into the inlet port. This represents a clever and simple way to continuously measure the state or conditions of the cell culture. For example, the sensor mounted in the inlet port can be adapted to measure the glucose level of the cell culture, and if the glucose level is low, the piston is actuated via a control mechanism, thus being pulled out and thereby able to replenish the cell culture chamber with fresh culture medium, including fresh glucose.

[0068] In another embodiment of the first aspect, at least one sensor is mounted on a portion of the at least one wall, such as the inner wall, of the container that is in direct contact with the cell culture chamber.

[0069] In another embodiment of the first aspect, the at least one wall, such as the outer wall, includes an inlet port leading to the fresh culture medium chamber. This enables the provision of fresh culture medium in the bioreactor, which is particularly useful at the start of use. The inlet port is directly connected to the fresh culture medium chamber and is preferably located at the periphery of the at least one wall, such as the outer wall, at the second end of the container, i.e., furthest from the cell culture chamber.

[0070] In another embodiment of the first aspect, the at least one wall, such as the outer wall, includes a port for introducing or removing air, liquid, cells, cell aggregates, or bioactive molecules (e.g., drugs). This entry port may be the same as the entry port described above leading to the fresh culture medium chamber. Preferably, the entry port is disposed on the circumferential side of the container or on a removable portion of the at least one wall, such as the outer wall, of the container.

[0071] The inlet port or any of the ports preferably has a cup surrounding it to facilitate keeping the area around the inlet port clean, dry and sterile and to facilitate the removal of air bubbles.

[0072] In another embodiment of the first aspect, an additional cell culture chamber is adapted to be connected in series with the cell culture chamber, the additional cell culture chamber having conduits for transferring the fresh culture medium and having holes for allowing the culture medium to flow from the cell culture chamber to the additional cell culture chamber. Thus, an additional cell culture chamber, preferably in the shape of a culture dish, is inserted between the removable end of the container (i.e., the container according to the first aspect) and the remainder of the bioreactor. This additional cell culture chamber has conduits for transferring the fresh culture medium from the remainder of the bioreactor to the removable end of the container and has holes or openings for allowing the culture medium to flow between the cell culture chamber and the additional cell culture chamber. In a particular embodiment, one or more additional cell culture chambers are assembled on or within the container by inserting additional cell culture chambers, preferably substantially in the shape of a culture dish. In another particular embodiment, at least one sensor is mounted on or within the one or more additional cell culture chambers, such that the contents of each chamber can be monitored independently.

[0073] For a series of connected cell culture chambers, it should be noted that the cell culture chambers at one or both ends (i.e., the first and / or the last) may include dual vents (as disclosed herein) located on the front or back rather than the top side port, while any cell culture chamber in the middle should include a top side port or any other suitable port to allow for exchange of air or gas with the outside of the bioreactor or its ambient air or gas for gas exchangers and humidifiers (if present).

[0074] In a second aspect, a bioreactor is provided (wherein at least one wall comprises an inner wall and an outer wall, the inner wall and the outer wall defining an annular compartment between the walls), wherein the cell culture chamber is located within the annular portion of the container (i.e., the annular compartment). Thus, a bioreactor suitable for rotation is provided, the bioreactor comprising:

[0075] Container, the container comprising:

[0076] A first end and a second end define a central axis of the container, the central axis extending along a first direction, such as the length direction, of the container from the first end to the second end;

[0077] At least one wall that extends along the first direction of the container;

[0078] At least one culture medium conduit, which defines a volume for receiving fresh culture medium;

[0079] The inner chamber is defined by at least a portion of a space confined within the at least one wall and includes a fresh culture medium chamber and a waste culture medium chamber;

[0080] The at least one culture medium conduit is or includes a cell culture chamber in fluid communication with the waste culture medium chamber and the fresh culture medium chamber;

[0081] A movable wall is configured within the interior to separate the fresh culture medium chamber from the waste culture medium chamber within the interior.

[0082] In some implementation schemes,

[0083] - The at least one wall includes an inner wall and an outer wall, the inner wall and the outer wall defining (or creating) an annular compartment between the walls.

[0084] - The at least one culture medium conduit is the annular compartment, and

[0085] - The interior chamber is defined by at least a portion, for example, all of the space confined within the interior wall.

[0086] In some implementation schemes,

[0087] -The at least one wall is a single wall, and

[0088] - The at least one culture medium conduit is at least one conduit arranged inside or outside the single wall.

[0089] Therefore, the cell culture chamber is annular (or included within the one or more culture medium conduits), meaning the cell culture chamber occupies the annular compartment or sub-compartment (or the one or more culture medium conduits). This allows for a more compact construction of the bioreactor and is well-suited for small cell culture chamber volumes. A larger average radial distance makes it easier to find the 'stationary orbit' rotational speed.

[0090] In a third aspect, a bioreactor system for growing cell cultures or tissues is provided, the system comprising:

[0091] A bioreactor suitable for rotation, the bioreactor comprising a container, the container comprising:

[0092] A first end and a second end, defining a central axis of the container, the central axis extending along a first direction, such as a length direction, of the container from the first end to the second end; at least one wall extending along the first direction of the container; at least one culture medium conduit defining a volume for receiving fresh or waste culture medium, preferably fresh culture medium;

[0093] The inner chamber is defined by at least a portion of a space confined within the at least one wall and includes a fresh culture medium chamber and a waste culture medium chamber;

[0094] The cell culture chamber is in fluid communication with the annular compartment and the fresh culture medium chamber or the waste culture medium chamber, preferably the culture medium chamber; and

[0095] A movable wall is configured within the interior to separate the fresh culture medium chamber from the waste culture medium chamber within the interior.

[0096] The movable wall is connected to a piston-shaped displacement element to cause the movable wall to be axially displaced along the first direction of the container;

[0097] The bioreactor system also includes:

[0098] - A retaining roller adapted to support the bioreactor and enable the bioreactor to rotate;

[0099] - A drive element, such as a drive wheel, for rotating the bioreactor;

[0100] - A retaining block for supporting the piston, which is connected to the movable wall via a piston shaft;

[0101] - A drive unit for moving the retaining block and thereby displacing the piston.

[0102] Preferably, the drive element and drive unit for rotating the bioreactor and for moving the retaining block are syringe pumps.

[0103] In some implementation schemes,

[0104] - The at least one wall includes an inner wall and an outer wall, the inner wall and the outer wall defining (or creating) an annular compartment between the walls.

[0105] - The at least one culture medium conduit is the annular compartment, and

[0106] - The interior chamber is defined by at least a portion, for example, all of the space confined within the interior wall.

[0107] In some implementation schemes,

[0108] -The at least one wall is a single wall, and

[0109] - The at least one culture medium conduit is at least one conduit arranged inside or outside the single wall.

[0110] In a fourth aspect, a bioreactor system for growing cell cultures or tissues is provided, wherein the cell culture chamber of the bioreactor is located in an annular portion, i.e., in the annular compartment. Therefore, a bioreactor system for growing cell cultures or tissues is provided, the system comprising:

[0111] A bioreactor suitable for rotation, the bioreactor comprising a container, the container comprising:

[0112] A first end and a second end, defining a central axis extending along a first direction, such as a length direction, of the container from the first end to the second end; an outer wall and an inner wall extending along the first direction of the container to create an annular compartment between the walls;

[0113] The inner chamber is defined by a space confined within the inner wall and includes a fresh culture medium chamber and a waste culture medium chamber;

[0114] The annular compartment is a cell culture chamber in fluid communication with the waste culture medium chamber and the fresh culture medium chamber;

[0115] A movable wall is configured within the interior to separate the fresh culture medium chamber from the waste culture medium chamber within the interior.

[0116] The movable wall is connected to a piston-shaped displacement element to cause the movable wall to be axially displaced along the first direction of the container;

[0117] The bioreactor system also includes:

[0118] A retaining roller, adapted to support the bioreactor and enable the bioreactor to rotate;

[0119] A driving element, such as a drive wheel, is used to rotate the bioreactor;

[0120] A retaining block for supporting the piston, the piston being connected to the movable wall via a piston shaft; and

[0121] A drive unit for moving the retaining block and thereby displacing the piston.

[0122] According to a fifth aspect, a bioreactor is provided for growing cell cultures and tissues, the bioreactor comprising:

[0123] - A cell culture chamber, configured with cell culture medium.

[0124] - A circumferential gas exchanger, which is arranged circumferentially around or along at least a portion of the cell culture chamber or around the central axis or longitudinal axis of the bioreactor, for example or preferably around a predetermined axis of rotation of the bioreactor, wherein the circumferential gas exchanger includes a cavity comprising a volume connecting the gas exchange interface of the cell culture chamber to the ambient air or gas of the bioreactor.

[0125] In this way, the gas exchanger is arranged off-center (but generally still around the center and / or axis of rotation) and away from the longitudinal axis (typically extending between the first and second ends and substantially parallel to the axis of rotation), meaning the gas exchanger is not 'stacked' next to the housing or any other components in the longitudinal direction. By having a circumferential gas exchanger, the longitudinal extent of the cell culture chamber arrangement is also significantly reduced, thereby reducing the longitudinal 'footprint' / form factor, which may be beneficial for design considerations.

[0126] It should be noted that even though the provision of such a circumferential gas exchanger works particularly well according to the first aspect, it can be used independently.

[0127] In some embodiments, the gas exchange interface is a porous or non-porous circumferentially permeable membrane, such as a semi-permeable membrane, configured to exchange gases such as oxygen and carbon dioxide with the contents of the cell culture chamber, wherein the circumferentially permeable membrane is arranged circumferentially along the circumferential portion of the cell culture chamber.

[0128] In some embodiments, the circumferentially breathable membrane is a connecting wall that connects a first end and a second end, wherein the first end, the second end, and the connecting wall at least partially define the cell culture chamber.

[0129] In some embodiments, the gas exchange interface or the circumferential breathable membrane is supported by at least one support structure, such as a grid-like support structure, the at least one support structure including a plurality of openings configured to connect the gas exchange interface or the circumferential breathable membrane to the air or gas in the cavity of the circumferential gas exchanger.

[0130] In some embodiments, the circumferential gas exchanger is connected to the ambient air or gas of the bioreactor via at least one gas or air inlet and / or outlet.

[0131] In some embodiments, the bioreactor is configured to rotate about a predetermined axis of rotation, and at least one of the at least one gas or air inlet and / or outlet is a dual vent or port configured to, in response to the rotation of the bioreactor about the predetermined axis of rotation, draw ambient air or gas into the cavity of the circumferential gas exchanger and discharge air or gas from the cavity of the circumferential gas exchanger, thereby generating an airflow.

[0132] In some embodiments, the bioreactor further includes:

[0133] - A circumferential humidifier, wherein the circumferential humidifier

[0134] The arrangement is situated around at least a portion of the cell culture chamber or around the central axis or longitudinal axis of the bioreactor, for example or preferably around a predetermined axis of rotation of the bioreactor.

[0135] The gas exchanger includes or is connected to one or more liquid or humidifying reservoirs or elements configured to humidify or moisturize air or gas in at least a portion of the cavity of the gas exchanger.

[0136] In some embodiments, the one or more liquid or humidifying reservoirs or elements are configured to humidify or moisturize at least a portion of air or gas adjacent to or near a gas exchange interface or breathable membrane.

[0137] According to a sixth aspect, a bioreactor suitable for rotation is provided, the bioreactor comprising:

[0138] Container, the container comprising:

[0139] - A first end and a second end, defining a central axis of the container, the central axis extending along a first direction, such as a length direction, of the container from the first end to the second end; at least one wall extending along the first direction of the container; at least one culture medium conduit defining a volume for receiving fresh or waste culture medium;

[0140] - An inner chamber, defined by at least a portion of a space confined within said at least one wall, and comprising a fresh culture medium chamber and a waste culture medium chamber;

[0141] - A cell culture chamber, which is in fluid communication with the at least one culture medium conduit and the fresh culture medium chamber and / or the waste culture medium chamber; and

[0142] - A movable wall configured within the interior to separate the fresh culture medium chamber from the waste culture medium chamber within the interior.

[0143] The bioreactor further includes a humidification system, which comprises:

[0144] - One or more liquid or humidifying reservoirs or elements

[0145] - Evaporation chamber, such as an evaporation labyrinth and a filter, the filter forming one of the walls of the cell culture chamber, and

[0146] - and liquid or moisture transport elements, such as wicks, configured to transport liquid or moisture from the one or more liquid or moisture reservoirs or elements to the evaporation chamber.

[0147] The humidified atmosphere will therefore come into direct contact with the filter, such as a semi-permeable membrane, which forms one of the walls of the culture chamber. Thus, gas can be exchanged relatively freely between the culture chamber and the atmosphere surrounding the equipment without net evaporation of liquid from the culture chamber.

[0148] In some embodiments, the humidification system is provided, for example, with a port for filling and refilling the liquid reservoir (e.g., with liquid, typically sterile water) and at least one open port connecting the evaporation chamber or the evaporation labyrinth to the outside atmosphere. In some embodiments, the humidification system is arranged between the cell culture chamber and the waste culture medium chamber. An outlet or valve for allowing culture medium (liquid culture medium) to enter the waste culture medium chamber from the culture chamber may, for example, pass through (without mixing) the liquid or humidification reservoir or element to the waste culture medium chamber from the center. As an alternative to using liquid, the humidifier may, for example, comprise one or more of the following: aqueous or dissolved materials, such as gels, sponges, particulate materials (e.g., water-beads, aqua-beads, slush powder, or hydrogel powder).

[0149] Any embodiment of the first aspect may be used in conjunction with the second, third, fourth, fifth, or sixth aspect of the invention. Any embodiment of the sixth aspect may be used in conjunction with any one or more of the other aspects. Attached Figure Description

[0150] Aspects of the invention are illustrated in the accompanying drawings, in which:

[0151] Figure 1 According to the embodiment of the bioreactor of the first aspect, the cell culture chamber is located at the first flat end of the container;

[0152] Figure 2 The internal portion of the container according to the first aspect is shown to demonstrate a simple way of constructing the bioreactor;

[0153] Figure 3 The implementation scheme of the bioreactor according to the first aspect includes a humidification system for the filter located between the cell culture chamber and the waste culture medium chamber;

[0154] Figure 4 According to the first aspect of the bioreactor implementation scheme, the filter humidification system is arranged along the circumference of the cell culture chamber, and wherein the humidification system is disposed between the cell culture chamber and the outside atmosphere;

[0155] Figure 5 An embodiment relating to cell culture chamber 30 and additional cell culture chamber 80 is shown;

[0156] Figure 6 It is an implementation scheme of the bioreactor according to the second aspect, wherein the cell culture chamber is the annular part of the container;

[0157] Figure 7A bioreactor system according to a third aspect is shown, the bioreactor system comprising the bioreactor itself, a drive element for rotating the bioreactor, and a drive unit for displacing a piston.

[0158] Figure 8 An alternative implementation scheme for the bioreactor according to the first aspect is illustrated schematically;

[0159] Figure 9 The diagram schematically shows a front view and a cross-sectional side view of an embodiment of a bioreactor as disclosed herein, including a circumferential gas exchanger and a circumferential humidifier.

[0160] Figure 10 A-10E schematically illustrate a front, first ('right') side, first cross-sectional (AA), second cross-sectional (CC), and third cross-sectional (BB) view of an exemplary embodiment of the cell culture chamber apparatus disclosed herein; and

[0161] Figure 11 schematic display Figure 10 An exploded perspective view of an exemplary embodiment of the cell culture chamber apparatus of A-10E. Detailed Implementation

[0162] refer to Figure 1 The bioreactor 10 includes a container 12, which includes an inner chamber comprising a fresh culture medium chamber 14 and a waste culture medium chamber 16. In this embodiment and corresponding embodiments, the container 12 further includes an inner wall 18 and an outer wall 20, both walls extending along a first direction of the container 12 (the length direction in the illustrated example) to create an annular compartment 22 between these walls, thereby forming a double-walled container. The annular compartment 22 can be a single compartment in the form of a single conduit, i.e., a single culture medium conduit, or multiple sub-compartments or multiple conduits, such as… Figure 2As shown in c. Container 12 includes: a first end 24, preferably a first flat end; and a second end 26, preferably a second flat end, located on the opposite side of container 12. The first end 24 and the second end 26 define a central axis 28, as indicated by a horizontal dotted line, extending along the length of the container from the first end 24 to the second end 26. Container 12 also includes: a cell culture chamber 30 disposed at the first end 24 of the container; and a movable wall 38 adapted within the inner chamber to separate a fresh culture medium chamber 14 from a waste culture medium chamber 16, while retaining at least a portion of the fresh and waste culture media (as indicated by the dotted areas) within the inner chamber. The cell culture chamber 30 may have an inlet orifice 32 and an outlet or valve 34, the inlet orifice allowing culture medium (liquid culture medium) to enter the cell culture chamber 30 from the annular compartment 22, and the outlet or valve allowing culture medium (liquid culture medium) to enter the waste culture medium chamber 16 from the culture chamber 30. Similarly, the annular compartment 22 has an inlet orifice 36 and an outlet orifice 32, the inlet orifice allowing culture medium from the fresh culture medium chamber 14 to enter the annular compartment 22, and the outlet orifice allowing liquid / culture medium from the annular compartment 22 to enter the cell culture chamber 30. The outlet orifice of the annular compartment corresponds to or coincides with the aforementioned inlet orifice 32 of the cell culture chamber 30. A movable wall 38 is connected to the piston 40 via a piston shaft 42. When the piston moves in the right direction as indicated by the arrow (in the orientation of the figures), the movable wall 38 is displaced, thereby allowing fresh culture medium from the fresh culture medium chamber 14 to pass through the annular compartment 22 into the cell culture chamber 30. The container 12 also includes a removable end 46 of the outer wall 20 (including the cell culture chamber 30) located at the first end 24 of the container. The container may also include, for example, a circumferential inlet port 74 (as shown) providing access to the cell culture chamber 30.

[0163] Figure 2 The internal parts of a bioreactor are shown, and the simple design and construction of a bioreactor are demonstrated by rapidly assembling such parts.

[0164] Figure 2 A shows an embodiment of an inner cylinder of a container, the inner cylinder including an inner wall 18 and a plurality of raised ridges 48, the plurality of raised ridges extending along a first direction of the inner wall 18 and thus along a first direction of the container 12 (the length direction in the illustrated example), and also each extending outward perpendicularly, i.e., in a radial direction perpendicular to the length direction, until contacting the outer wall 20. The space between the raised ridges 48 defines one or more sub-compartments 22' within the annular compartment 22, such as Figure 2 As shown in the side view presented in c. Figure 2a also shows hole 50, access port 52 ( Figure 2 b) Access to the waste culture medium chamber 16 through the hole (hole).

[0165] Figure 2 b shows the outer cylinder of the container, which includes an outer wall 20. The outer wall 20 is removable and includes an access port 52 adapted to mate with a hole 50 in the inner wall 18. Figure 2 a). The inlet port 52 also functions as a pressure equalization port to, for example, alleviate pressure differentials. The outer wall 20 includes an inlet port 54 (and a plug) for allowing fresh culture medium to be introduced into the fresh culture medium chamber 14. A locking flange 56 is also provided, which corresponds to a flange 58 or 60 of the cell culture chamber 30 or the additional cell culture chamber 80. Figure 5 (a) or (b) connection.

[0166] Figure 2 c is the cross-section when the inner cylinder is assembled into the outer cylinder. The parallel raised ridges 48 create multiple annular sub-compartments or conduits 22'.

[0167] Figure 2 Figure d shows the interior portion of the container in the form of a movable wall 38, which is connected to the piston 40 via a piston shaft 42. When assembling the bioreactor, the piston shaft 42 passes through a hole 44 in the flat end 26 of the outer wall.

[0168] Figure 3 A bioreactor according to a first aspect as disclosed herein is shown, wherein a humidification system is inserted between a cell culture chamber and a waste culture chamber. The humidification system, i.e., the humidifier, includes a liquid reservoir 62, an evaporation chamber such as an evaporation labyrinth 68, and a filter 72. Liquid (typically sterile water) is filled into the liquid reservoir 62 via port 64. This liquid is then conveyed via a wick 66 to the evaporation labyrinth 68, where it humidifies the atmosphere of the evaporation chamber. The evaporation labyrinth 68 has at least one open port 70 leading to the atmosphere surrounding the apparatus, through which gases, such as oxygen and carbon dioxide, can be exchanged. The humidified atmosphere is in direct contact with a filter, such as a semi-permeable membrane 72, which forms one of the walls of the culture chamber 30. In this way, gases can be exchanged relatively freely between the culture chamber 30 and the atmosphere surrounding the apparatus without net evaporation of liquid from the culture chamber 30. In practice, water is lost from the liquid reservoir 62 at a very approximate rate of 1-2 mL / week. Therefore, for a 10 mL bioreactor as described herein, the reservoir 62 should have a volume of 10-20 mL. The filter 72, the evaporation labyrinth 68, and the liquid reservoir 62 have conduits that connect the cell culture chamber to the waste culture medium chamber.

[0169] Figure 4An embodiment of a bioreactor according to the first aspect is shown, wherein the humidification system has a filter in the form of a semi-permeable membrane 72' arranged circumferentially around the cell culture chamber 30, i.e., along the circumferential portion of the cell culture chamber. The humidification system includes the filter 72', a liquid reservoir 62' containing sterile water, an additional filter such as a wick or permeable filter 66', and an evaporation chamber in the form of an evaporation labyrinth 68'. The additional filter 66' will be highly permeable to allow evaporation from the liquid reservoir 62' into the evaporation labyrinth 68'. Therefore, in this embodiment, it would be simpler to... Figure 3 The wick 66 of the embodiment is replaced with a breathable filter 66' that has different properties from the semi-permeable membrane 72'. The humidification system is also provided with a port 64 for refilling the liquid reservoir 62' and at least one open port 70' connecting the evaporation labyrinth 68' to the outside atmosphere. For access to the cell culture chamber 30 of the bioreactor, as already in, for example... Figure 1 Port 74 is arranged as described in the text.

[0170] Figure 4 The implementation scheme of the bioreactor shown is preferred over Figure 1 The embodiment shown, due to the presence of a humidification system, will significantly enhance gas exchange between the cell culture chamber 30 (or additional culture chamber 80) and the surrounding atmosphere, and will reduce or eliminate water loss from said cell culture chamber 30 or 80. The difference is significant, enabling bioreactors to be typically used in incubators without additional humidification. This is a major advantage as it will reduce the risk of infection in the incubator.

[0171] Therefore, the circumferential gas exchanger and circumferential humidification system are easy to set up. It should be noted that the circumferential gas exchanger (and circumferential humidification system) can be set up independently of the other features of the first aspect. Please also refer to... Figure 9 And related descriptions of alternative implementations of the circumferential gas exchanger and circumferential humidification system as disclosed herein, based on the fifth aspect.

[0172] For at least three reasons, Figure 4 The implementation scheme of the bioreactor shown is also preferred over Figure 3 As shown in the diagram. First, it is easier to regulate gas exchange via plugs or sliding covers with ports 70' that can be partially or completely closed, allowing for regulation of the gas exchange of individual bioreactors rather than having to regulate the atmosphere of the entire incubator. A second reason is that the bioreactor shown is more compact along axis 28 and ultimately less expensive to manufacture. Furthermore, the circumferential gas exchangers (and circumferential humidifiers) are also advantageous for multiple cell culture chambers / additional cell culture chambers (30, 80) connected in series, as they do not obstruct the connection in the first / length direction.

[0173] Figure 5 A cell culture chamber 30 is shown, including a circumferential inlet port 74 and a planar inlet port 76. These ports provide access to cell cultures within the cell culture chamber 30 and may be equipped with sensors for monitoring cell culture conditions (e.g., glucose levels). A corresponding flange 56 is also provided. Figure 2 b) or 78 Figure 5 b) Engaging locking flange 58.

[0174] Figure 5 b illustrates an additional cell culture chamber 80, which can be inserted between the first cell culture chamber 30 and a double-walled container containing waste growth medium and fresh growth medium. The additional cell culture chamber 80 contains orifices or openings 82 for allowing culture medium, such as liquid medium, to flow between the two cell culture chambers 30 and 80. Therefore, these cell culture chambers can be considered as arranged in series. A conduit 84 is also arranged to allow liquid to flow through conduit 22 and via (without mixing) at least one additional cell culture chamber 80 between the fresh medium chamber 14 and the cell culture chamber 30. Therefore, more additional cell culture chambers can be arranged therein. For a series of connected cell culture chambers, it should be noted that cell culture chambers at one or both ends (i.e., the first and / or the last) may include dual vents located on the front or back rather than the top port 74 (see, for example, 140 elsewhere), while any cell culture chamber in the middle should include the top port 74 or any other suitable port.

[0175] Figure 6 This is an embodiment of the bioreactor 10” as disclosed in the second aspect herein, wherein the cell culture chamber 30 is an annular portion of the container 12. Therefore, the annular compartments 22, 22' are the cell culture chamber 30 and are in fluid communication with the waste culture medium chamber 16 and the fresh culture medium chamber 14. This results in a more compact configuration. The container 12 is preferably provided with an arrangement 86 for opening and closing the bioreactor. For clarity, inlet ports, sensor locations, and humidification systems have been omitted, but they can certainly be included in a similar manner as described above. Such embodiments can also be combined with a circumferential gas exchanger (and, for example, a circumferential humidifier) ​​according to the fifth aspect and as disclosed herein.

[0176] Figure 7 A bioreactor system 88 according to a third aspect disclosed herein is shown. The system can be mounted on a platform 90 and includes a bioreactor 10 as disclosed herein, the bioreactor comprising a cell culture chamber 30 at a first flat end of a double-walled vessel 12. Alternatively, [further details are needed]. Figure 4 Example of a bioreactor implementation scheme 10' Figure 6Example of bioreactor implementation scheme 10 and Figure 8-11 The example of bioreactor implementation scheme 100 replaces Figure 1-3 The example of a bioreactor implementation 10 is shown. Retaining rollers 92 and 94, etc., are respectively positioned below and above the container 12 to support the bioreactor 10 and / or enable the bioreactor 10 to rotate. A drive element in the form of a drive wheel 96 is attached to the container 12 and thus to the bioreactor 10, thereby providing rotation. This function can also be performed by 92 and / or 94. The system also includes a retaining block 98 for supporting a piston 40, the system comprising a piston shaft 42 and a movable wall 38, and a drive unit 500 for moving the retaining block 98. Optional conduits 502 (such as tubes) are provided, extending, for example, inside the piston shaft 42 through the central axis of the piston to the outside 504. This allows for direct and simple sampling of cells from the cell culture chamber 30.

[0177] Figure 8 Alternative embodiments of the container for a bioreactor according to the first aspect are schematically illustrated. An embodiment of container 12 for a bioreactor as disclosed herein (see, for example, 10, 10', 10" in other figures) is shown, the container corresponding to... Figure 1 The containers, except as described below. For clarity, not all elements are again in [the container]. Figure 8 As shown in the image. Figure 8 A shows a first end view of container 12. Figure 8 B shows the perspective side / top view of container 12, and Figure 8 C shows a longitudinal cross-sectional view of container 12.

[0178] Similar to Figure 1 As disclosed herein, the container includes a first end 24, a second end 26, a waste culture medium chamber 16, a fresh culture medium chamber 14, and a movable wall 38 connected to the piston shaft 42 of the piston 40 and separating the waste culture medium chamber 16 and the fresh culture medium chamber 14, as described and disclosed herein.

[0179] Figure 8 The container in A-8C is shown without a cell culture chamber, but can be used, for example, with... Figure 1 Cell culture chamber (see, for example) Figure 1 30) of the cell culture chambers and particularly those including, as disclosed herein and for example, Figure 8 The removable end of the cell culture chamber shown in D (see example) Figure 1 and 3 Use it together with 46).

[0180] like Figure 8As shown in C, container 12 does not include the outer and inner walls defining the annular compartment (or sub-compartment) (see example). Figure 1 Container 12 (as shown in 18 and 20) comprises at least one single wall 18' (i.e., not double-walled) and at least one culture medium conduit 22' replacing the annular compartment or sub-compartment. In some embodiments in which the container is generally cylindrical, container 12 comprises a single generally cylindrical wall 18'.

[0181] At least one culture medium conduit 22' is fluidly connected to the fresh culture medium chamber 14 via a corresponding orifice or valve 36' through the (single) wall 18', thereby allowing (liquid) culture medium to flow from the fresh culture medium chamber 14 to the corresponding culture medium conduit 22' when the movable wall 38 is properly moved. Each culture medium conduit 22' also includes (typically arranged at the opposite end or the other end of the orifice 36') an additional orifice or valve 32, which is connected to the... Figure 1 The inlet orifice or valve of the cell culture chamber (see, for example) Figure 1 32) Align, correspond, or coincide (but the size of the cell culture chamber may differ slightly from the specified size). Figure 1 In this case, it is aligned with the culture medium conduit 22' rather than the annular compartment or sub-compartment. The first end 24 of the container 12 also includes an orifice 34 or alternatively a valve or the like for allowing (liquid) culture medium to flow from the culture chamber into the waste culture medium chamber 16, wherein the orifice or valve 34 is aligned with the outlet or valve of the cell culture chamber (see, for example...). Figure 1 34) Alignment, correspondence, or overlap.

[0182] In some implementation schemes and such particularly Figure 8 As shown in Figure A, at least one culture medium conduit 22' is arranged externally to wall 18' on the outer surface of the main portion of container 12, adjacent to or on the outer surface of wall 18'. In an alternative embodiment, at least one culture medium conduit 22' is arranged internally to wall 18' on the inner surface of wall 18', adjacent to or on the inner surface of wall 18'. In principle, a mixture of internal and external culture medium conduits 22' may also be present.

[0183] Figure 8 The container 12 is shown as comprising only a single culture medium conduit 22', but in other embodiments, the container 12 may include multiple culture medium conduits 22'. In embodiments having multiple culture medium conduits 22', they may be equidistantly positioned, for example, around the (circular) circumference of a single wall 18' (inside and / or outside).

[0184] Figure 8 D shows a product with a removable lid or similar (such as...) Figure 1The container 12 (in the middle) includes a cell culture chamber 30, which is attached to one end of a hole or valve 32 aligned with a culture medium conduit 22'.

[0185] Figure 9 A front view (shown on the right) and a cross-sectional side view (shown on the left) are schematically shown according to some embodiments and an embodiment of a bioreactor 100 disclosed herein, said embodiments including a circumferential gas exchanger and a circumferential humidifier according to the fifth aspect. Hereinafter and herein, "bioreactor" is equivalently referred to as a 'cell culture chamber apparatus'.

[0186] The illustration (see both views) shows a cell culture chamber apparatus 100 as disclosed herein. The cell culture chamber apparatus 100 includes a housing 30 as disclosed herein (also referred to hereinafter and herein as a cell culture chamber; see also 30 in other figures, for example), defined by at least one connecting wall 18' of a first end 111, a second end 112, and connecting ends 111, 112. The housing 30 is, for example, constructed of a shell / main shell 105, wherein the shell / main shell 105 is cylindrical (by way of example) and includes the housing 30 at its center (by way of example). In the illustrated and corresponding embodiments, at least one connecting wall 18' is formed (e.g., supported) by a circumferentially permeable membrane 72' arranged along or as a circumferential portion of the housing 30 (i.e., its perimeter or a portion thereof) and configured for exchanging gases, such as oxygen and carbon dioxide. The circumferentially permeable membrane 72' may, for example, correspond to a combination Figure 4 The membrane (72') shown and illustrated is, for example, a semipermeable membrane.

[0187] Humidification of the atmosphere near or adjacent to the circumferential breathable membrane 72' typically reduces or prevents evaporation of the cell culture medium and, for some cell culture media, can further greatly promote gas exchange through the circumferential breathable membrane 72'. Cells produce CO2, which combines with water in solution to form bicarbonate (which is acidic). Humidification of the atmosphere causes the outer surface of the circumferential breathable membrane 72' to become moist, and this promotes the escape of CO2 from the culture medium and, in this case, slows down the acidification process. This process occurs in cell culture types that do not rely on CO2 to buffer the culture medium (e.g., those containing HEPES, zwitterionic sulfonic acid buffers). The most widely used types of growth media rely on bicarbonate in the medium and CO2 in the atmosphere to buffer the pH of the medium. Here, humidification of the outer surface of the circumferential breathable membrane also promotes the 'capture' or 'release' of CO2, thereby improving the pH stability of the culture medium. Humidification can be provided by the cell culture chamber device 100 located in a humidified incubator or by a humidifier as described below.

[0188] As shown in the front view (on the right side of the figure), the cell culture chamber apparatus 100 includes a gas exchange inlet and outlet for a gas exchanger for the cell culture chamber apparatus 100, which can be any suitable inlet, conduit, etc. Preferably, and as shown, the gas exchange inlet and outlet are in the form of a dual vent or similar 140, thereby connecting the circumferential gas exchanger to the outside of the cell culture chamber apparatus 100 or its ambient air or gas. Alternatively, the gas exchange inlet and outlet are similar to Figure 4 70' in the diagram. In the illustrated embodiment, as an example, the dual vents 140 are located on the front or front side of the housing or main housing 105 (see also later figures) or the like. In this particular (and corresponding embodiment), the gas exchanger includes a permeable membrane 72' configured to exchange gases, such as oxygen and carbon dioxide, with the contents (e.g., cell culture medium) of the housing 30 / housing. Specifically, oxygen can be supplied to the housing 30, and carbon dioxide can be removed from the housing 30. In the illustrated and corresponding embodiments, the membrane 72' constitutes at least one connecting wall 18' or one or more portions thereof of the housing 30.

[0189] Gas exchange inlet and outlet / dual vent 140 is fluidly connected to membrane 72', thereby connecting membrane 72' to the outside of the cell culture chamber apparatus 100 or its ambient air or gas. In at least some embodiments, dual vent 140 is configured according to Coanda The effect or principle of operation. In such embodiments, a wall or other suitable barrier 151 (indicated in the figure by a straight dashed line) is located between the two corresponding vents of the dual vent 140, thereby separating and sealing them from each other at that location, i.e., separating and sealing them in the shortest direction between them in this particular instance. However, the two vents of the dual vent 140 are fluidly connected to each other via another path inside the housing 105 of the cell culture chamber device 100, and are also fluidly connected to at least a portion of the gas exchange membrane 72', for example, via one or more conduits, open spaces, cavities, etc. When the cell culture chamber device 100 is rotated counterclockwise, ambient air or gas is drawn into and drawn out of the cell culture chamber device 100 through the dual vent 140, as by Figure 9As indicated by the arrows in the front view and cross-sectional side view, during counterclockwise rotation, air or gas is drawn into the cell culture chamber apparatus 100 more specifically through the left (in the front view) vent 140 (as indicated by the arrows from black to gray) and discharged to the outside of the cell culture chamber apparatus 100 through the right (in the front view) vent 140 (as indicated by the arrows from gray to black), thereby creating an internal airflow 310 with a direction indicated by the light gray dashed circular arrow. This is the case for counterclockwise rotation. If the cell culture chamber apparatus 100 rotates clockwise, the direction of the airflow 310 inside the housing 105 will be reversed due to symmetry, i.e., the light gray dashed circular arrow will be clockwise, and air or gas will be drawn in through the right vent 140 and discharged through the left vent 140.

[0190] In this way, an effective airflow 310 is readily provided to contact the ambient gas or air of the membrane 72' and the cell culture chamber apparatus 100, thereby conveniently adding oxygen to / from the membrane 72' and thus to / from the contents of the housing 30 and removing carbon dioxide.

[0191] In some further embodiments, the degree of air movement or flow 310 can be adjusted, for example, by using a slider or small or different sized plugs or by adjusting the corresponding size of the opening of the vent of the dual vent 140 in another suitable manner.

[0192] In some further embodiments (and as shown in the figures), the cell culture chamber apparatus 100 optionally includes a circumferential humidifier or humidifying or moisturizing element or system (equivalently referred to herein as a humidifier) ​​62' configured to humidify or moisturize the air or gas adjacent to at least a portion of the membrane 72' (which is also presently a portion thereof). The humidifier will significantly enhance gas exchange between the contents of the housing 30 and the ambient air or gas, and further reduce or eliminate water or liquid loss from the housing 30 when containing liquid or aqueous solutions. The effects are significant, enabling the cell culture chamber apparatus 100 to be generally used in incubators without additional humidification. This is advantageous because it generally reduces the risk of infection in incubators and also simplifies incubator operation.

[0193] In some such embodiments, the circumferential humidifier 62' includes (or is connected to) one or more liquid or humidifying reservoirs or elements. It is advantageous if the weight distribution of the circumferential humidifier 62' is at least somewhat uniform around the central axis or axis of rotation of the cell culture chamber apparatus 100. It is also advantageous if such one or more liquid or humidifying reservoirs or elements have or provide a relatively large surface area for evaporation.

[0194] There are several advantageous possibilities for humidifying or moisturizing the air or gas at least adjacent to the membrane 72' (at least a portion thereof).

[0195] In some embodiments, the circumferential humidifier 62' includes an element or reservoir containing (preferably sterile) liquid water or other moisturizing liquid (see, for example...). Figure 3 , 4 62, 62'), the element or reservoir, for example, has one or more suitable filters, an outlet, and an additional (breathable and particularly semi-breathable) membrane (see 62', 62') that connects water or liquid with airflow 310. Figure 4 The airflow 310 is humidified or moisturized by means of elements such as 66', etc. The element or reservoir may be, for example, a single circumferential unit or alternatively several separate and different units (e.g., uniformly distributed around a central axis and / or a rotation axis).

[0196] In an alternative embodiment, the circumferential humidifier 62' includes one or more of an aqueous or dissolved material, such as gel, sponge, or particulate material (e.g., water droplets, aquatic beads, slush powder, or hydrogel powder, etc.), which readily provides evaporation of water or liquid and effectively influences the airflow 310. Such a solid humidifying or moisturizing element may be supported or secured in the housing 105, for example, by a (open) shell, wall, or other support structure (e.g., 145 in the following figure), or, for example, supported or secured in the housing 105 to a (open) shell, wall, or other support structure (e.g., 145 in the following figure).

[0197] In the case of water droplets or gels, these can be fixed, adhered, glued, etc. to the inner wall of the main housing / outer housing 105 (as mentioned, for example, or preferably uniformly around the central axis and / or rotation axis), whereby a support structure is not necessary.

[0198] For implementations that do not include water or liquid reservoirs (such as water droplets, gels, etc. as mentioned above), this substance can be directly positioned in conduits, cavities, open spaces, etc., including the airflow 310, thereby greatly increasing the humidification or moisturizing effect on the airflow 310 and enabling a reduction in the overall space / footprint of the cell culture chamber apparatus 100. This avoids the need for Figure 4 and 9 Individual storage devices, such as 62'.

[0199] It should be noted that for implementations without a humidifier (e.g., for use in a humidified incubator or otherwise), the cell culture chamber device 100 shown will not include the circumferential humidifier 62' shown and can therefore have a reduced size.

[0200] Figure 10A-10E schematically show a front, first ('right') side, first cross-sectional (AA), second cross-sectional (CC), and third cross-sectional (BB) view of an exemplary embodiment of a cell culture chamber apparatus as disclosed herein.

[0201] Figure 10 Figure A shows a front view of an exemplary preferred embodiment of a cell culture chamber apparatus as disclosed herein. The front of the cell culture chamber apparatus 100 is shown. In this particular (and corresponding embodiment), the bottom, base, or wall of the cover 102 (see also, for example...) Figure 11 102) constitutes the first end 111 (or a portion thereof or window 113) of the cell culture chamber apparatus 100. The bottom surface or base of the cover 102, together with the first or central outer shell 101, forms a shell as disclosed herein (see, for example...). Figure 1 , 3 30 and 5A and 7 Figure 9 (30) as will become more apparent from some of the following figures. In some implementations (and as... Figure 10 As shown in C, 10D, 10E, 11, etc., the first or central housing 101 includes a central cavity for (e.g., or preferably releasably) receiving at least a portion of the cover 102 and more particularly (in the illustrated and corresponding embodiments) receiving the bottom surface or base of the cover 102. The first or central housing 101 and the cover 102 may, for example, include corresponding releasable fastening elements (such as snap-fit, pin, friction fit, etc.) to releasably secure them together. Alternatively, they may be non-releasably fixed to each other or, for example, integrally formed.

[0202] As an example, Figure 10 The cell culture chamber device 100 of A-10E is basically cylindrical in shape with a circular first end.

[0203] In this particular (and corresponding) embodiment, the central housing 101 further includes a gas exchange loop, element, or system (not shown; see example) in the form of a circumferential gas exchange system including a circumferentially permeable membrane. Figure 4 72' and Figure 9 , 10 (62' and 72' in D and 11). As mentioned, the bottom surface or base of the cover 102 constitutes the first end 111 (or a portion thereof or window 113) of the cell culture chamber apparatus 100.

[0204] In this particular (and corresponding) embodiment, the central housing 101 also includes, for example, as disclosed herein and in combination with... Figure 4Or the circumferential humidifier described in 9 (not shown). Some alternative embodiments of the cell culture chamber apparatus 100 do not include any humidifier, for example, used in a humidified incubator or otherwise.

[0205] The central housing 101 optionally includes a gas exchange inlet and outlet for a gas exchanger as disclosed herein (see [link]). Figure 9 , 10 (and 130, 140, 151, 310, etc. in 11), which takes the form of a double vent 140 located in front of the central housing 101. It has been combined, for example... Figure 9 The dual vent 140 is described in more detail.

[0206] Further indications are those marked AA (e.g.) Figure 10 As shown in C), BB (as shown in C) Figure 10 (as shown in E) and CC (as shown in E) Figure 10 The three sections shown in D).

[0207] In some embodiments (and as shown), the cell culture chamber apparatus 100 also includes a closable and / or sealable (first) port 76, which provides a user with access to the interior of the housing, for example, to remove samples (e.g., remove spheroids), empty or fill the housing, etc. In the illustrated embodiment, the closable and / or sealable port 76 includes a conduit (from the interior of the housing to the exterior of the cell culture chamber apparatus 100) and, for example, a simple plug or the like 160. The port may (alternatively or additionally) advantageously be located at the top of the cell culture chamber apparatus 100, as this can, for example, avoid or reduce bubble formation by allowing overflow. Such a 'top-side' port is, for example, as... Figure 1 , 3 74 is shown in 10 and 11.

[0208] In some embodiments (and as shown), the cell culture chamber apparatus 100 further includes one or more reference and / or identification markers, here identification code 155 and reference marker 180. Identification code 155 is preferably unique to a particular cell culture chamber apparatus 100. Reference marker 180 is capable of determining the orientation of the cell culture chamber apparatus 100. Reference and / or identification markers 155, 180 are preferably machine-readable, for example, by a suitable imaging or vision unit or system. In some embodiments, the cell culture chamber apparatus further includes one or more alignment elements (e.g., positioning strips and slits or slots, etc.) for aligning different parts of the cell culture chamber (ensuring or facilitating that one part is received only by another part in proper orientation) (e.g., properly aligning cover 102 with the first or central housing 101). Reference marker 180 may be, for example, such an alignment element.

[0209] Therefore, a very compact (longitudinal) cell culture chamber device 100 is provided, particularly due to the circumferential gas exchange system and (if present) the circumferential humidifier.

[0210] Optionally, the cover 102 includes a plurality of level or fill rate indicators 190, thereby easily indicating the actual volume of cell culture medium contained in the housing.

[0211] In some embodiments and as shown in the figure, the cell culture chamber device 100 also includes one or more (here, two) feet, upright elements 501, etc., so that the cell culture chamber device 100 can stand upright and not roll. This can be achieved by using easier or more reliable ports, inlets, etc. (see, for example, port 74 herein).

[0212] Figure 10 B shows the view from the side and from right to left (according to...) Figure 10 (A's orientation) observation Figure 10 A side view of a cell culture chamber apparatus 100. The cell culture chamber apparatus 100 shown includes a main housing 105 that receives (e.g., permanently or in a fixed manner) a central housing 101, which in turn receives (e.g., releasably) a cover 102. Further shown is a (second) port 74 (or more precisely its plug or valve 170), which is fluidly connected to the housing and provides (additional) access to the housing.

[0213] First range / length (in) Figure 10 (in the left and right directions of B) and the second range / height or diameter (in Figure 10 The ratio between B (in the vertical direction) is about 1 to about 3-4, for example about 1 to about 3.5, but can be different, for example as disclosed herein with respect to other embodiments.

[0214] Figure 10 C demonstrates as shown by Figure 10 The first cross-sectional view given by A's AA shows a housing 30, defined by a first end 111 as the bottom or base of a cover 102, a second end 112 as the bottom or base of a central housing 101, and sidewalls of the cavity of the central housing 101. Further shown is a main housing 105, which receives the central housing 101 and the cover 102 in a very compact manner.

[0215] As mentioned, the second port 74 provides access to the housing 30 (in addition to the first port 76). (As in example...) Figure 9 As described, the dual vents 140 connect the external air or gas of the cell culture chamber apparatus 100 or its ambient air to the circumferential gas exchange system (see, for example...). Figure 9 , 10Connect to 130, 140, 310, etc. in 11.

[0216] As can be seen, the closable and / or sealable first port 76 and its conduit connect the interior of the housing 30 to the exterior of the cell culture chamber apparatus 100. The port wall is part of the cover 102, thereby allowing easy access to the contents of the housing 30. In a similar manner, access to the interior of the housing 30 is provided via the second port 74 (with a plug 170). The plug wall of 74 / 170 is part of the central outer shell 101. Note that the first port 76 and the second portion 74 are arranged on different sides of the cell culture chamber apparatus 100, allowing easy access to the housing from several different sides of the cell culture chamber apparatus 100.

[0217] Further shown is the gas exchange inlet and outlet in the form of a dual vent 140, as previously described.

[0218] Figure 10 C's view is from left to right (according to...) Figure 10 (In the orientation of A) the center of observation is vertically cut.

[0219] Figure 10 D shows, as demonstrated by Figure 10 The CC of A is given as a second cross-sectional view viewed from right to left.

[0220] Similarly, housing 30, first transparent end 111, transparent or translucent second end 112, central housing 101, cover 102, closable and / or sealable ports 76 and 74, and main housing 105 are shown.

[0221] A portion of the permeable membrane 72' of the circumferential gas exchange system and the grid-like structure 130 of the circumferential humidifier is further shown.

[0222] It also demonstrates, according to some embodiments of a circumferential humidifier, elements for retaining and / or supporting water, liquid, or moisture (in combination with...) Figure 9 (Further explanation) Wall structural elements or the like 145.

[0223] In some embodiments, the cell culture chamber apparatus 100 optionally includes one or more markers 115—in the form of multiple concentric circles 115, which can provide the user with some fixed markings relative to which to observe the slow movement of the contained spheres. The markers 115 (by example) are arranged on the 'outer' side of the second end 112.

[0224] Figure 10 The view of D is offset from the center, shifted to the left, and from right to left (according to...). Figure 10 Vertical cut observed in orientation A)

[0225] Figure 10 E shows, as demonstrated by Figure 10 The third cross-sectional view given by BB of A.

[0226] The illustration shows a housing 30, a first end 111, a second end 112, a cover 102, a closable and / or sealable port 76, and two wall structure elements or the like 145 for retaining and / or supporting water, liquid, or moisturizing elements according to some embodiments.

[0227] Figure 10 E's view is from top to bottom (according to...) Figure 10 (Observed in the orientation of A) Horizontal center cut.

[0228] Figure 11 schematic display Figure 10 An exploded perspective view of an exemplary embodiment of the cell culture chamber apparatus of A-10E.

[0229] The exploded diagram is shown below. Figure 10 A-10E components.

[0230] Figure 11 The grid-like structure 130 and the breathable membrane 72' of the circumferential humidifier are shown more clearly. In the assembled state of the cell culture chamber device 100, the breathable membrane 72' is adjacent to and positioned on the inner side of the grid-like structure 130, with a first end 111 opposite to a second end 112, and the second end 112 aligned with the opening 185 of the main housing 105, thereby allowing easy inspection of the contents of the housing from that side when the second end 112 is transparent. Another port 150 is further shown, which is aligned with the first port 74 in the assembled state.

[0231] In at least some embodiments, the main housing 105, the central housing 101 (and thus the second end 112), and the cover 102 (and thus the first end 111) may be, for example, made of the same material, such as as disclosed elsewhere herein.

[0232] like Figure 10 and 11 The embodiment of the cell culture chamber apparatus 100 shown provides a very compact (particularly in the longitudinal direction) self-contained and fully functional cell culture chamber apparatus 100 or bioreactor, wherein the gas exchanger (and humidifier (if included)) is arranged away from the central axis and / or the axis of rotation. Furthermore, the cell culture chamber apparatus 100 has a petri dish-like design, thereby enabling easy and familiar manipulation.

Claims

1. A bioreactor (10, 10', 100) suitable for rotation, said bioreactor comprising: Container (12), said container (12) comprising: - A first end (24) and a second end (26), the first end (24) and the second end (26) defining a central axis (28) of the container (12) extending along a first direction of the container (12) from the first end (24) to the second end (26); at least one wall (18, 18', 20), the at least one wall (18, 18', 20) extending along the first direction of the container (12); at least one culture medium conduit (22, 22'), the at least one culture medium conduit (22, 22') defining a volume for receiving fresh or waste culture medium; - An inner chamber, the inner chamber being defined by at least a portion of a space confined within the at least one wall (18, 18', 20) and comprising a fresh culture medium chamber (14) and a waste culture medium chamber (16). - A cell culture chamber (30), said cell culture chamber (30) being in fluid communication with the at least one culture medium conduit (22, 22') and the fresh culture medium chamber (14) and / or the waste culture medium chamber (16); and - A movable wall (38) is configured in the inner chamber to separate the fresh culture medium chamber (14) from the waste culture medium chamber (16) in the inner chamber. The cell culture chamber (30) is separately arranged at the first end (24) of the container (12) and is provided with an inlet hole (32) and an outlet hole or valve (34), the inlet hole (32) being used to allow culture medium to enter the cell culture chamber (30) from the at least one culture medium conduit (22, 22'), and the outlet hole or valve (34) being used to allow culture medium to enter the waste culture medium chamber (16) from the culture chamber (30).

2. The bioreactor (10, 10', 100) according to claim 1, wherein... - The at least one wall (18, 18', 20) includes an inner wall (18) and an outer wall (20), the inner wall (18) and the outer wall (20) defining an annular compartment between the walls (18, 20). - The at least one culture medium conduit (22, 22') is the annular compartment, and - The interior chamber is defined by at least a portion of the space confined within the interior wall (18).

3. The bioreactor (10, 10', 100) according to claim 1, wherein... - The at least one wall (18, 18', 20) is a single wall (18'), and - The at least one culture medium conduit (22, 22') is at least one conduit (22') arranged inside or outside the single wall (18').

4. The bioreactor (10, 10', 100) according to claim 2, wherein the at least one culture medium conduit (22, 22') is provided with an inlet hole and an outlet hole, the inlet hole being for allowing culture medium to enter the at least one culture medium conduit (22, 22') from the fresh culture medium chamber (14), the outlet hole being for allowing culture medium to enter the cell culture chamber (30) from the at least one culture medium conduit (22, 22'), and wherein the outlet hole of the annular compartment corresponds to the inlet hole of the cell culture chamber.

5. The bioreactor (10, 10', 100) according to claim 4, wherein the at least one culture medium conduit (22, 22') is the annular compartment or the at least one conduit (22').

6. The bioreactor (10, 10', 100) according to claim 1, wherein at least a portion of the at least one wall (18, 18', 20) is removable to provide access to the cell culture chamber (30).

7. The bioreactor (10, 10', 100) according to claim 6, wherein at least a portion of the at least one wall (18, 18', 20) is removable and serves as a removable end (46) of the at least one wall (18, 18', 20) located at the first end (24) of the container (12).

8. The bioreactor (10, 10', 100) according to claim 1, wherein at least a portion of the at least one wall (18, 18', 20) is transparent glass or plastic, configured to allow observation of the culture medium, cells and spheroids contained therein.

9. The bioreactor (10, 10', 100) according to claim 2, wherein the inner wall (18) has a plurality of raised ridges (48) extending along the first direction of the container (12) and extending vertically outward until contacting the outer wall (20), and wherein the space between the raised ridges (48) defines one or more sub-compartments within the annular compartment.

10. The bioreactor (10, 10', 100) according to claim 1, wherein the container (12) has a cylindrical or generally cylindrical shape and is adapted to rotate about a rotation axis by one or more associated rotating elements, the rotation axis being the central axis (28) extending along the first direction of the container (12).

11. The bioreactor (10, 10', 100) according to claim 1, wherein the movable wall (38) is connected to the displacement element (40) to axially displace the movable wall (38) along the first direction of the container (12).

12. The bioreactor (10, 10', 100) according to claim 11, wherein the displacement element (40) is a piston, and the movable wall (38) is connected to the piston via a piston shaft (42) that coincides with the central axis (28) extending along the first direction of the container (12).

13. The bioreactor (10, 10', 100) according to claim 12, wherein a conduit (502) is provided, the conduit (502) extending from the cell culture chamber (30) through the center of the piston shaft (42) to at least the outside (504) of the container (12).

14. The bioreactor (10, 100) according to claim 1, wherein the container (12) or a portion thereof is made of breathable plastic, or The container (12) includes a breathable membrane (72, 72') for gas exchange, the breathable membrane (72') being arranged along the circumferential portion of the cell culture chamber (30).

15. The bioreactor (10, 100) according to claim 14, wherein the gas comprises oxygen and carbon dioxide.

16. The bioreactor (10, 10', 100) according to claim 14, wherein a humidification system is disposed between the culture chamber (30) and the outside atmosphere, the humidification system comprising an evaporation chamber (68, 68') of a liquid reservoir (62, 62') and a filter.

17. The bioreactor (10, 10', 100) according to claim 16, wherein the liquid reservoir (62, 62') contains sterile water.

18. The bioreactor (10, 10', 100) according to claim 16, wherein the liquid reservoir (62, 62') is an evaporation labyrinth.

19. The bioreactor (10, 10', 100) according to claim 16, wherein the filter is the breathable membrane (72') arranged along the circumferential portion of the cell culture chamber (30).

20. The bioreactor (10, 10', 100) according to claim 16 or 19, wherein the humidification system further comprises an additional filter (66') arranged in fluid communication with the liquid reservoir (62, 62') and the evaporation chamber (68, 68').

21. The bioreactor (10, 10', 100) according to claim 20, wherein the additional filter (66') is arranged along the circumferential portion of the container (12) and is located between the liquid reservoir (62, 62') and the evaporation chamber (68, 68').

22. The bioreactor (10, 10', 100) according to claim 1, wherein the cell culture chamber (30) includes at least one inlet port (74, 76).

23. The bioreactor (10, 10', 100) according to claim 22, wherein the cell culture chamber (30) further comprises a sensor.

24. The bioreactor (10, 10', 100) according to claim 23, wherein the sensor is removably mounted in the inlet port.

25. The bioreactor (10, 10', 100) according to claim 1, wherein at least one wall (18, 18', 20) includes an inlet port (54) leading to the fresh culture medium chamber.

26. The bioreactor (10, 10', 100) according to claim 1, wherein an additional cell culture chamber (80) is adapted to be connected in series with the cell culture chamber (30), the additional cell culture chamber (80) having conduits for transferring the fresh culture medium and having pores (82) for allowing the culture medium to flow from the cell culture chamber (30) to the additional cell culture chamber (80).

27. The bioreactor (10, 10', 100) according to claim 1, wherein the bioreactor (10, 10', 100) comprises at least one membrane for preventing at least a portion of the cell culture in the cell culture chamber (30) from leaving the cell culture chamber (30).

28. The bioreactor (10, 10', 100) according to claim 27, wherein the at least one membrane is disposed above one or more of the inlet and / or outlet orifices of the cell culture chamber (30).

29. The bioreactor (10, 10', 100) according to claim 1, wherein one or more additional cell culture chambers (80) are assembled on or in the container (12) by inserting additional additional cell culture chambers (80).

30. The bioreactor (10, 10', 100) according to claim 29, wherein the additional cell culture chamber (80) is a cell culture chamber in the shape of a culture dish.

31. The bioreactor (10, 10', 100) according to claim 29, wherein at least one sensor is installed on or in one or more of the additional cell culture chambers (80) to allow independent monitoring of the contents of each chamber.

32. The bioreactor (10, 10', 10'', 100) according to claim 14, wherein the bioreactor (10, 10', 10'', 100) further comprises: - A circumferential humidifier, wherein the circumferential humidifier Arranged around at least a portion of the cell culture chamber (30) or around the longitudinal axis of the bioreactor (10, 10', 10'', 100), and Includes or is connected to one or more liquid or humidifying reservoirs or elements, said one or more liquid or humidifying reservoirs or elements being configured to humidify or moisturize gas in at least a portion of the cavity of the gas exchanger (130, 140, 151, 310).

33. The bioreactor (10, 10', 10'', 100) according to claim 32, wherein the longitudinal axis of the bioreactor (10, 10', 10'', 100) is the central axis of the bioreactor (10, 10', 10'', 100).

34. The bioreactor (10, 10'', 100) according to claim 32, wherein the one or more liquid or humidifying reservoirs or elements are configured to humidify or moisturize at least a portion of the gas adjacent to or adjacent to the gas exchange interface or the permeable membrane (120).

Images