CN220149584U

CN220149584U - Device and bioreactor for culturing cells

Info

Publication number: CN220149584U
Application number: CN202320757275.4U
Authority: CN
Inventors: B·海伦; A·休伯特; B·迈雷斯; C·杜蒙; G·贝尔哈伊; J-C·德拉蒙德
Original assignee: Univolcels Technologies Inc
Current assignee: Univolcels Technologies Inc
Priority date: 2022-04-07
Filing date: 2023-04-07
Publication date: 2023-12-08
Anticipated expiration: 2033-04-07
Also published as: CN117070351A

Abstract

Apparatus and bioreactor for culturing cells. The apparatus for culturing cells includes a bioreactor including a housing having a plurality of removable parts for modular construction and easy assembly or disassembly. The removable parts include a fixed bed and a support for the fixed bed. More than one support may be provided. The plurality of supports may interlock with each other or with the cover of the bioreactor. The support is interlocked with the housing to prevent relative rotation. The plurality of supports includes a support frame for supporting the fixed bed and allowing fluid to flow through the support frame. The fixed bed forms a peripheral chamber between the support and the housing to hold the fixed bed. One or more probes and catheters may be inserted into the bioreactor to test parameters of the fluid or to add or remove fluid from the interior.

Description

Device and bioreactor for culturing cells

Cross Reference to Related Applications

The present application claims priority from U.S. provisional patent application Ser. No. 63/328,461 submitted at 7 of 4 of 2022, ser. No. 63/330,967 submitted at 14 of 4 of 2022, ser. No. 63/410,252 submitted at 27 of 9 of 2022, and Ser. No. 63/412,709 submitted at 3 of 10 of 2022, the disclosures of which are incorporated herein by reference. The present application also relates to U.S. provisional patent applications serial nos. 62/758,152, 62/733,375, and 62/608,261, each of which is incorporated herein by reference. The disclosures of U.S. patent application publication No. 2018/0282678, international patent application PCT/EP2018/076354, U.S. provisional patent application 62/711,070, and U.S. provisional patent application 62/725,545 are also incorporated herein by reference.

Technical Field

This document relates generally to the field of cell culture and more particularly to modular bioreactors, bioreactor systems, and in particular to apparatus and bioreactors for culturing cells.

Background

Bioreactors are often used to culture cells. Typically, bioreactors include complex housing arrangements formed from multiple parts that require excessive welding or gluing to assemble. This increases the complexity of manufacture, the risk of failure, and ultimately the cost to the end user.

Another problem relates to the ability to maximize cell density in the cell growth area of the bioreactor. Many past proposals for fixed bed bioreactors use packed beds. While such packed beds may well promote cell growth and provide certain advantages, the volume of space in the bioreactor required to produce such beds is significant. While achieving desired cell growth, ease of expanding bioreactors with unstructured packed or fluidized beds is also challenging, and there is currently a need for bioreactors that can be used under various operating conditions in the art, including, for example, within sterile hoods, where gaps may be limited.

Cell culture can be expensive in terms of both time and resources involved in a given process. Failure of such efforts or suboptimal conditions for a given process may result in waste of time and resources, or at least suboptimal use of available resources, including researcher time, physical components, and various materials involved in the given process. For this reason, small scale operations may be useful, such as for process development, experimental design, assessing the feasibility of a given process, or for optimizing parameters thereof. If such an assessment is made on a larger scale, each run of a given process for experimental purposes utilizes a significant amount of resources.

Furthermore, assessing process parameters in a single bioreactor system requires either additional time to run the experimental sequence or additional physical resources to control multiple individual single bioreactor systems. For example, evaluating the effect of a change in a given parameter (e.g., temperature, pH, DO, oxo/kLa, etc.) or a given set of parameters for a given process may require a significant amount of time for sequential operation in order to change the parameter and/or a significant amount of resources and space for operating the process at the same time under a variety of different parameters in order to optimize the process.

Thus, there is a need for an improved bioreactor that is easy and inexpensive to manufacture and easy to expand to accommodate a variety of operating conditions. The bioreactor can be assembled in a highly repeatable manner. Once the optimal conditions and/or parameters are determined, the associated bioreactor system can also facilitate expansion.

Disclosure of Invention

In a first embodiment, an apparatus for culturing cells is disclosed, comprising: a bioreactor comprising a housing having walls forming an interior compartment; a fixed bed for culturing cells; a fixed bed support for a fixed bed, the fixed bed support adapted to be removably positioned within the interior compartment of the housing; and one or more locators for uniformly spacing the fixed bed support from the wall of the housing.

In this or other embodiments, the fixed bed support includes a central portion and a peripheral portion extending radially outward from the central portion and including one or more locators, the peripheral portion having an outer diameter corresponding to an inner diameter of the housing. The peripheral portion may include one or more protrusions. The peripheral portion may comprise an outer ring connected to the central portion by one or more protrusions.

In this or other embodiments, the peripheral portion includes an annular disc-shaped surface having a plurality of apertures therein. The peripheral portion may comprise a mesh or screen. The peripheral portion may be adapted to support the fixed bed from below. The peripheral portion may be adapted to allow fluid to flow therethrough.

In another aspect of this or other embodiments, the central portion may include a separator that forms a plurality of spaces in an interior of the central portion for receiving the conduit within the housing. The plurality of spaces may have different sizes. At least one of the conduits may be adapted to convey a fluid flow to or from the interior of the central portion. The separator may be adapted to locate at least one of the conduits, the conduits being adapted to convey the fluid flow away from the inner wall of the central portion to prevent contact between the conduits and falling film fluid flowing down the inner wall.

In another aspect of this or other embodiments, the central portion of the support forms a receptacle for receiving the agitator. The agitator may comprise an impeller adapted to rotate about an agitator support, the agitator support being adapted to receive and retain the fixed bed support. The agitator support may comprise a tubular post connected to a flexible drain connected to the lid of the bioreactor.

In another aspect of this or other embodiments, the peripheral portion of the fixed bed support can include one or more protrusions in the form of a plurality of radially extending arms. The plurality of radially extending arms may be connected to a rim having an outer diameter corresponding to the inner diameter of the housing. A plurality of radially extending arms can engage and support the fixed bed.

In another aspect of this or other embodiments, the housing includes a receiver for receiving at least one of the one or more protrusions. The central and peripheral portions of the fixed bed support may comprise a single unitary structure.

In another aspect of this or other embodiments, the fixed bed comprises one or more layers of woven or nonwoven material wrapped around a central portion of the fixed bed support.

In another aspect of this or other embodiments, the apparatus can further include a seal for sealing between the inner wall of the housing and the fixed bed support.

In another aspect of this or other embodiments, the fixed bed includes a plurality of fixed bed sections, and the fixed bed support further includes a plurality of interlocking support sections for supporting each of the plurality of fixed bed sections. Each interlocking support portion may be adapted to interlock with an adjacent support portion. The apparatus may further comprise a first seal for sealing each adjacent interlocking support section together. The apparatus may further comprise a second seal for sealing each of the plurality of portions to an inner wall of the housing.

In another aspect of this or other embodiments, the apparatus further comprises an upper frame for positioning over the fixed bed. The upper frame may have a sufficient height to form at least one pocket for allowing fluid to accumulate therein upon exiting the upper end of the fixed bed, and a sensor for sensing a characteristic of the fluid in the at least one pocket. The upper frame may be adapted to engage a cover of the bioreactor. The upper frame may form a plurality of pockets having different volumes. The upper frame may be adapted to receive or align one or more samplers for sampling the fixed bed.

In another aspect of this or other embodiments, the device further comprises a cover removably connected to the housing. The cover may be adapted to hold the fixed bed vertically in place within the housing. The cover may include an overhang portion for engaging a fixed bed or fixed bed support. The cap may be adapted to threadably engage the housing. The device may further comprise a gasket between the cover and the housing.

In another aspect of this or other embodiments, the apparatus may include a port in a wall of the housing that is above an upper end of the fixed bed when positioned therein.

In another aspect of this or other embodiments, the housing may comprise a one-piece rigid structure that forms the interior compartment.

In another embodiment of the present disclosure, an apparatus for culturing cells is disclosed, comprising: a housing and a lid together defining a container having an interior compartment; and an assembly for positioning within the interior compartment, the assembly comprising a fixed bed adapted to culture cells, wherein the assembly is adapted to interlock with the container to maintain the position of the fixed bed within the interior compartment.

In one aspect of this or other embodiments, the assembly includes an upper portion adapted to interlock with the lid. The upper portion may include an upper frame for positioning over the fixed bed. The upper frame may have a sufficient height to form at least one pocket for allowing fluid to accumulate therein upon exiting the upper end of the fixed bed, and a sensor for sensing a characteristic of the fluid in the at least one pocket. The upper frame may include a recess for engaging a protrusion extending from the cover of the bioreactor. The upper frame may form a plurality of pockets, each having a different volume.

In another aspect of this or other embodiments, the fixed bed includes a plurality of fixed bed sections, each fixed bed section being associated with one of a plurality of supports adapted to interlock with an adjacent support. The upper frame may be adapted to interlock with at least one of the plurality of supports. Each of the plurality of supports may include a central portion and a peripheral portion, the peripheral portion adapted to allow fluid to reach the fixed bed, the peripheral portion having an outer diameter corresponding to an inner diameter of the housing.

In another aspect of this or other embodiments, the apparatus may further comprise at least one O-ring located between at least two of the plurality of supports or between the cover and at least one of the plurality of supports. The cover may be adapted to provide downward pressure on the assembly when the cover is attached to the housing. The downward pressure may be sufficient to maintain the at least one O-ring in place without glue.

In another aspect of this or other embodiments, the assembly may include a lower portion for receiving the agitator, the lower portion being adapted to interlock with at least one support for supporting the fixed bed.

In a third embodiment of the present disclosure, an apparatus for culturing cells is described, comprising: a bioreactor comprising a housing having walls forming an interior compartment; a fixed bed for culturing cells; and a support for the fixed bed, the support being adapted to be removably positioned within the interior compartment, the support at least partially forming a chamber housing the agitator and including one or more centering projections extending toward a wall of the housing.

In one aspect of this or other embodiments, the projections engage a fixed bed.

In another aspect of this or other embodiments, the housing includes one or more receptacles for receiving one or more protrusions. The engagement between the one or more protrusions and the one or more receptacles may be adapted to prevent rotation of the support within the housing.

In a fourth embodiment, an apparatus for culturing cells is disclosed, comprising: a housing having walls forming an interior compartment; a fixed bed for culturing cells within the interior compartment; one or more probes for extending into the interior compartment, adjacent to or into the fixed bed; and an upper frame covering the fixed bed for holding the fixed bed and organizing the one or more probes.

In one aspect of this or other embodiments, the upper frame includes one or more indicia for indicating the position or orientation of the one or more probes.

In another aspect of this or other embodiments, one or more probes are attached to the upper frame.

In a fifth embodiment, an apparatus for culturing cells is disclosed, comprising: a one-piece housing; at least one fixed bed for culturing cells; and a plurality of fixed bed supports adapted to interlock to be positioned within the single piece housing.

In one aspect of this or other embodiments, each of the plurality of fixed bed supports includes an annular portion and a support frame extending radially outward from the annular portion. The support frame may have an outer diameter corresponding to the inner diameter of the housing. The support frame may include a generally planar extension. The support frame may be adapted to allow fluid to flow therethrough. The support frame may extend from the bottom of the annular portion. The support frame may include a plurality of radially extending arms connected to the peripheral ring. The support frame comprises a mesh or screen. The support frame may be adapted to support the at least one fixed bed from below. The support frame may be adapted to serve as a base that positions the fixed bed on the fixed bed support.

In another aspect of this or other embodiments, each of the fixed bed supports includes a protrusion or receiver for interlocking with a corresponding protrusion or receiver on an adjacent fixed bed support of the fixed bed support.

In a sixth embodiment, an apparatus for culturing cells is disclosed, comprising: a bioreactor comprising a housing having walls forming an interior compartment; a fixed bed for culturing cells; an annular fixed bed support for supporting a fixed bed, the fixed bed support adapted to be removably positioned within the interior compartment of the housing and defining a peripheral chamber between the annular fixed bed support and the housing; an impeller for circulating a fluid through the fixed bed within the peripheral chamber; and an impeller support for extending at least partially through the impeller and centering the impeller in the housing; wherein the impeller is attached to the impeller support by a snap fit connection.

In a seventh embodiment, a method of manufacturing a fixed bed bioreactor is disclosed that includes the step of interlocking one or more fixed bed supports within a single piece housing.

In one aspect of this or other embodiments, the interlocking step includes interconnecting the first fixed bed support to the second fixed bed support.

In another aspect of this or other embodiments, the method further comprises the step of positioning the impeller within a portion of the first fixed bed support.

In another aspect of this or other embodiments, the interlocking step includes interlocking the second fixed bed support with a cover for covering the housing.

In yet another aspect of this or other embodiments, the method further comprises the step of forming a first seal between the first fixed bed support and the second fixed bed support.

In another aspect of this or other embodiments, the method further comprises the step of forming a second seal between the second fixed bed support and the housing.

In another aspect of this or other embodiments, the method further comprises the step of wrapping the fixed bed around each of the fixed bed supports.

In an eighth embodiment, a bioreactor for culturing cells is disclosed. The bioreactor includes a housing having walls defining an interior compartment, a plurality of fixed beds for culturing cells, and a plurality of annular fixed bed supports. Each of the plurality of fixed bed supports is adapted to support a respective at least one of the plurality of fixed beds. Each of the plurality of fixed bed supports includes an annular section and a support frame extending radially outward from the annular section. The support frame has an outer diameter corresponding in size to an inner diameter of a wall of the housing, the support frame being adapted to support at least one of the plurality of fixed beds from below and to allow fluid to flow through the support frame. The plurality of fixed bed supports are adapted to interlock with one another to form a peripheral chamber between the plurality of annular fixed bed supports and the wall of the housing and a central chamber within the annular section.

The bioreactor further comprises: a cover for connecting to the housing and for sealing the plurality of fixed beds and the plurality of fixed bed supports in the interior compartment; a plurality of probes extending into the interior compartment adjacent to or into at least one of the fixed beds; an upper frame covering the plurality of fixed bed supports and forming a plurality of pockets for allowing fluid to accumulate therein upon exiting the upper ends of the plurality of fixed beds. At least one of the plurality of probes is adapted to sense a characteristic of the fluid in a respective one of the plurality of bags.

The bioreactor further comprises: an impeller for circulating a fluid within the bioreactor; and a container for accommodating the impeller. The container comprises: a plurality of openings adapted to allow fluid to flow from within the container to the peripheral chamber; and a plurality of locators in the form of radially extending arms extending therefrom and adapted to locate the container within the housing and to space the container from the walls thereof. The upper frame is adapted to interlock with at least one of the plurality of annular fixed bed supports and is adapted to interlock with the cover to prevent relative rotation therebetween.

In a ninth embodiment, a bioreactor for culturing cells is disclosed. The bioreactor comprises: a housing having walls defining an interior compartment; a removable fixed bed for culturing cells; a removable fixed bed support adapted to support a fixed bed. The fixed bed support is annular in shape and includes a plurality of arms extending radially outwardly, the radially extending arms defining an outer diameter corresponding in size to an inner diameter of the wall of the housing for positioning and centering the fixed bed support within the housing. The plurality of arms are adapted to support the fixed bed from below. The fixed bed support forms a peripheral chamber between an outer wall of the fixed bed support and the housing, and a central chamber within the fixed bed support. The fixed bed is adapted to be positioned within the peripheral chamber. The housing includes one or more receptacles in a wall of the housing for receiving at least one of the plurality of arms, the one or more receptacles adapted to support the fixed bed support within the interior compartment and prevent relative rotation of the fixed bed support within the housing.

The bioreactor further comprises: a cover for connecting to the housing and for sealing the fixed bed and fixed bed support in the interior compartment; and at least one probe extending into the interior compartment at a location within the peripheral chamber above the fixed bed.

The bioreactor further comprises: an impeller adapted to rotate on an impeller support, the impeller for circulating a fluid within the bioreactor. The impeller is located in a chamber formed between the lower portion of the fixed bed support and the floor of the housing. The impeller is adapted to circulate fluid outwardly from the central chamber to the peripheral chamber of the fixed bed support and upwardly through the fixed bed therein.

The bioreactor further comprises: a drain connected to the impeller support for draining fluid from the bioreactor.

Drawings

The features of the present disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:

fig. 1 is a perspective view of a first embodiment of a bioreactor according to one aspect of the present disclosure.

FIG. 1A is a cross-sectional view of the bioreactor of FIG. 1.

FIG. 2 is an exploded view of the bioreactor of FIG. 1.

Figure 3 shows a spiral fixed bed that may be used in conjunction with a bioreactor.

Fig. 3A, 3B and 3C show specific details of a spiral fixed bed.

Fig. 3D, 3E, 3F and 3G illustrate alternative arrangements for forming a structured fixed bed.

Fig. 4 and 4A show a vessel containing an agitator in the bioreactor of fig. 1.

Fig. 4B and 4C illustrate an impeller assembly for a bioreactor according to one aspect of the present disclosure.

Fig. 5 is an exploded perspective view of a fixed bed and a support for the fixed bed according to one aspect of the present disclosure.

Fig. 6 is a perspective view of a support having the fixed bed of fig. 5.

Fig. 6A is a bottom view of the support of fig. 5.

Fig. 7 is a side view of the bioreactor of fig. 1.

Fig. 7A is a cross-sectional view taken along line 7A-7A of fig. 7.

Fig. 8 is a perspective view of a frame for providing upper support for a fixed bed in accordance with an aspect of the present disclosure.

FIG. 9 is a cross-sectional perspective view of the bioreactor of FIG. 1.

Fig. 9A is an enlarged view of a portion of the bioreactor of fig. 9.

Fig. 10 is a partial cross-sectional view of another embodiment of a bioreactor according to another aspect of the present disclosure.

Fig. 10A is an exploded perspective view of the bioreactor of fig. 10.

Fig. 11 is an exploded perspective view of another embodiment of a bioreactor according to another aspect of the present disclosure.

Fig. 11A is a top view of the bioreactor of fig. 11.

FIG. 11B is a cross-sectional side view of the bioreactor taken along line 11B-11B of FIG. 11A.

FIG. 12 is another side view of the bioreactor of FIG. 11.

Fig. 12A is a cross-sectional view taken along line 12A-12A of fig. 12.

FIG. 13 is a partial cross-sectional view of the lower portion of the bioreactor of FIG. 11.

FIG. 14 is a cross-sectional view of the bioreactor of FIG. 11.

Fig. 15 is a top view of the bioreactor of fig. 11.

Fig. 15A is a cross-sectional view taken along line 15A-15A of fig. 15.

Fig. 15B is a partially enlarged view of fig. 15A.

Fig. 16 is a flowchart showing an assembly step of a bioreactor according to the present disclosure.

FIG. 17 is a schematic diagram of a bioreactor system.

FIG. 18 is a schematic representation of a bioreactor cell density sampling location.

FIG. 19 shows the pH and DO parameters in the bioreactor over time.

Figure 20 shows the metabolite levels in different sized bioreactors over time.

Detailed Description

Referring now to fig. 1, 1A and 2, one embodiment of a bioreactor 100 for culturing cells according to one aspect of the present disclosure is shown together. Bioreactor 100 includes an outer shell or housing 112, shown as transparent in fig. 1, to allow viewing of the internal structure. The housing 112 forms an interior compartment in which cell culture may be accomplished using various components or techniques, as further outlined in the description below.

In accordance with one aspect of the present disclosure, in some embodiments, the housing 112 may form a vessel including a one-piece or unitary structure, such as a can or tub having an open top. Providing such a vessel may eliminate the cost and complexity of forming the housing 112 from multiple components secured together, such as using welding or adhesives. Furthermore, such a configuration avoids the need for an associated hermetic seal in the body of the housing 112, thereby eliminating the possibility of leakage and/or contamination and improving the integrity of the bioreactor.

The manufacture of this one-piece housing 112 may include the use of injection molding techniques, three-dimensional printing, or other methods such that no seams are present in order to minimize exposure to contamination. In some applications, the housing 112 may be translucent or transparent. In other applications, the housing 112 may be opaque and may be made of any material, but if desired, plastic is preferred to allow for a single use arrangement.

A lid or cover 114 may overlie the open top of the housing 112 to cover or seal the interior compartment thereof. In one embodiment, the cover 114 is designed to be easily removed, such as by interlocking engagement with the housing 112 (which may include friction fit or bayonet fit), but removable fasteners, such as tabs and/or clips, clamps, and/or screws that may interlock with each other, may also be used. This helps to open the bioreactor 100 and may avoid the need to use a sampler (which tends to increase cost due to size limitations and may be difficult to implement in a particularly small vessel). The housing 112 and the cover 114 may together comprise a container for housing the remaining elements of the bioreactor.

The cover 114 may include various openings or ports P with removable closures or caps C for allowing selective introduction or removal of materials, fluids, gases, probes, sensors, samplers, etc., and increased design flexibility. In particular, the lid 114 may include a holder 114b, such as for receiving a suitable sensor (e.g., temperature, capacitance, permittivity, biomass, metabolite such as glucose or lactate, pressure, flow measurement, liquid level, pH or DO probe, etc.). As best shown in fig. 1A, an internal connector 114c for a conduit or pipe forms part of the cap 114. The cover 114 may further include a corresponding connector 114d for the medium extraction tube T. As shown in fig. 1 and 9A, a removable cap 114e with a suitable seal (such as an O-ring) may allow for auxiliary access if desired. A sampling port, such as in the form of a probe, for receiving a sampler may also optionally be provided in the cover 114.

In the interior compartment formed by the housing 112, several compartments or chambers receive and transport streams of fluid, gas, or both throughout the bioreactor 100. As shown in fig. 1A, in some embodiments, the chamber may include a first chamber 116 located at or near a base of the bioreactor 100. In some embodiments, this first chamber 116 may include an agitator for inducing fluid flow within the bioreactor 100. In some embodiments, the agitator may be in the form of a "plug-in" rotatable non-contact magnetic impeller 118, which thus forms the centrifugal pump 100 in the bioreactor. Instead of such an impeller 118, the agitator may also be in the form of a stirring rod, an external pump forming part of the fluid circulation system, or any other means for inducing fluid circulation within the bioreactor.

The agitation provided causes the fluid to flow upward (as indicated by arrow V in fig. 1A) into a second chamber, which may be a peripheral chamber 120 formed in and extending along an outer or peripheral portion of the bioreactor 100. Alternatively, the bioreactor 100 may be adapted to allow fluid flow in the opposite direction.

In some embodiments, bioreactor 100 is adapted to accommodate any form of cell culture bed 122, including packed beds, fixed beds, structured fixed beds, fluidized beds, and the like. For example, fig. 3 shows a fixed bed 122 in the form of a structured helical bed that may contain and retain growing cells in use. In some embodiments, the spiral bed may be in the form of a cartridge that may be dropped or placed into the peripheral chamber 120, and may be part of a support that forms such a cartridge (and is adapted to interlock with additional modular structures as further outlined in the description below). The bed 122 may be pre-installed in the housing 112 during factory manufacture prior to shipment to the end user or installed at the point of use of the end user.

The fluid exiting the second peripheral chamber 120 is transferred to the headspace formed by the upper chamber 121 on one side (upper side) of the bed 122 where the fluid is exposed to a gas, such as oxygen. The fluid may then flow radially inward to the third central chamber 126 to return to the lower portion of the bed 122. In some embodiments, this central chamber 126 may be cylindrical in nature, formed from one or more non-porous conduits or tubes 128 (which may include multiple annular portions of fixed bed support, each annular portion including a portion of a fixed bed, as further outlined below), and the flow may be such that a waterfall-like arrangement is formed. The central chamber 126 returns the falling or otherwise entering fluid to the first base chamber 116 (arrow R shows the return path) for recirculation through the bioreactor 100, creating a continuous loop (in this scenario "bottom to top", but this could be reversed or otherwise modified without departing from the present disclosure).

As perhaps best understood from the exploded view of fig. 2, an upper frame 130, such as a chuck, may also be provided. The upper frame 130 can be positioned above an upper end portion of the fixed bed 122, such as below the cover 114 and adjacent to the cover 114. The upper frame 130 can be used to organize structures extending from the cover 114 into the fixed bed 122 or near the fixed bed 122, such as one or more probes 132 in communication with the port P. In one aspect, probes or other structures extending from the cover may contact or attach to a portion of the upper frame 130. In another aspect, the probes or other structures may be spaced apart from the upper frame to avoid contact with the upper frame. If present, these probes 132 can be used to sense conditions and/or obtain samples from within the bioreactor 100, particularly the fixed bed 122, in order to determine cell growth characteristics if desired, without removing the cover 114.

Fig. 3 illustrates one embodiment of a matrix material for use as a structured fixed bed 122, particularly a spiral bed, in a bioreactor of the present disclosure. In some embodiments, one or more cell immobilization layers 122a may be adjacent to one or more optional spacer layers 122b, and the spacer layers 122b may comprise a woven or nonwoven mesh structure. In some embodiments, layering may optionally be repeated several times to achieve a stacked or layered configuration.

The network structure contained in spacer layer 122b forms a tortuous path for cells (see cell L in fig. 3A suspended or entrapped in the material of fixed layer 122a, and cell culture may form part of any of the inventions claimed herein) and fluid to flow through the channels created thereby when laminated between two fixed layers 122 a. Due to this type of arrangement, increased uniformity of cells is maintained within the structured fixed bed. In some embodiments, other spacer structures forming such tortuous paths may be used.

As shown in fig. 3, 3A, and 3B, the structured fixed bed 122 may be rolled helically or concentrically along an axis or core (e.g., a conduit or tube 128, which may be provided in multiple components). In some embodiments, the layers of the structured fixed bed are firmly wrapped, but may be loosely wrapped. In some embodiments, the diameter of the open center portion for receiving tube 128, the length of the layers, and/or the number will ultimately define the size of the assembly or substrate. In some embodiments, the thickness of each of the layers 122a, 122b may be between 0.1 millimeters and 5 millimeters, between 0.1 millimeters and 10 millimeters, or between 0.001 millimeters and 15 millimeters.

In some embodiments, other structures that form such tortuous paths may be used. For example, fig. 3D illustrates that one or more cell fixation layers 122a may be suitable for forming a structured fixed bed 122. One or more layers 122a provide a curved flow path (arrow B) from a linear or regular inflow (arrow a) without the use of additional spacer layers (although such spacer layers may be used if desired). This may be achieved, for example, by providing a layer of textile fibers or filaments 123, 125 that disrupt flow.

Fig. 3E shows that such results can be achieved using a woven or nonwoven material as the cell immobilization layer 122 a. This may be achieved by forming this layer 122a as a mesh arrangement with openings 127 (e.g. by 3D (three-dimensional) printing), through which openings 127 the fluid may pass and return again, thereby forming a tortuous path again promoting uniformity and also serving to further shear or split any bubbles present in the fluid. This function can be achieved again with or without the added spacer layer.

The orientation of the structured fixed bed 122 may be different from that shown in the bioreactor 100 as shown in fig. 1, 1A and 2, wherein the flow is vertically arranged (bottom to top in the example provided, see arrows V and R). For example, as shown in fig. 3F, bioreactor 100 may include a chamber 120 with a structured fixed bed 122 formed of one or more horizontally disposed layers of material. As shown in fig. 3D and 3E, one or more layers may comprise woven or mesh material, but as shown in fig. 3F, one or more cell fixation layers 122a (three are shown, but any number may be present) are sandwiched by adjacent spacer layers 122b (vertical spacing is exaggerated for purposes of illustration), which is optional. Thus, the flow is arranged from side to side (left to right or right to left), with the material layers (spacers or otherwise) providing channels for creating a tortuous flow (arrow B) from a linear or regular inflow (arrow a). The pumping action may be provided by an agitator or other pump at the inlet end of the chamber 120, and a return path is provided at the outlet end as schematically shown by path R. If desired, additional spacer layers may extend between the cell immobilization layers 122 a.

In another possible embodiment, and referring to fig. 3G, the structured fixed bed 122 comprises a three-dimensional (3D) monolithic matrix 124 in the form of a scaffold or grid formed of a plurality of interconnected cells or objects 124a, the plurality of interconnected cells or objects 124a having a surface for cell adhesion. Preferably, the matrix comprises a tortuous path for fluid and cells to flow therethrough in use. In these or other embodiments, the matrix may be in the form of a 3D (three-dimensional) array, lattice, scaffold, or sponge. In all cases, the substrate 124 may be disposable in nature to avoid the costs and complexities involved with cleaning according to biological treatment standards.

Referring to fig. 1, 1A, 4 and 4A, the bioreactor 100 of this first exemplary embodiment may include a support for supporting a fixed bed. In one form, this support may include a receptacle 140, the receptacle 140 for receiving an agitator, such as the impeller 118, in the interior compartment of the housing 112. The vessel 140 may be adapted to receive fluid from the central opening 142 and to eject fluid radially outward via one or more openings 144 (e.g., four openings spaced 90 degrees apart), such as a result of movement (rotation) of an agitator, such as the impeller 118.

The container 140 may also include one or more outward protrusions that act as locators for centering or uniformly spacing the container from the inner wall of the housing 112, but not attached to the inner wall of the housing 112. For example, the vessel 140 may include one or more radially extending arms 140a along an upper portion. These arms 140a may be adapted for aligning or centering the vessel within the housing 112 of the bioreactor 110 when the vessel rests on a surface (such as a floor) of the bioreactor 110. While the arm 140a may be on the container 140, the arm may alternatively be attached to an inner wall of the housing 112 and extend toward the container, but not attached to the container, for ease of removal.

Fig. 4A and 4B illustrate that the impeller 118 is adapted to rotate about a support, which may take the form of a tubular post 148. In one aspect, the tubular post 148 may be attached to the bottom of the housing 112. In another aspect, the tubular post 148 may be removable from the housing 112. The upper portion of this post 148 may include one or more vanes 150 and may be connected to a conduit 151. The conduit 151 may comprise a flexible tube and may be used either to supply gas to the container 140 or as a drain to withdraw fluid therefrom. In the latter case, the lower portion of the post 148 adjacent the base plate may be scalloped or provided with openings 148a to allow fluid flow. According to fig. 1A, a conduit 151 may be connected at an opposite or upper end to the cap 114 in fluid communication with one of the ports P.

As further shown in fig. 4B, the lower end of the post 148 may be located in a central opening 118a in the impeller 118. The post 148 may include a first or upper stop, such as a flange 148b, to limit the distance that the associated conduit 151 may travel when coupled to the upper end of the post. A second lower stop, such as flange 148c, may provide an upper limit for movement of the impeller 118. As shown, these stops are considered optional and may take forms other than flanges.

To facilitate low friction rotation, the bearing assembly 162 may support the impeller 118. In one example, the bearing assembly 162 may include a base 162a, the base 162a including a race 162b for receiving a bearing 164, the bearing 164 being in the form of, for example, a ball bearing, a cylindrical bearing, a bushing material, or any other bearing element suitable to facilitate relative movement between impellers. It will be appreciated that the impeller 118 may have a similar but inverted seat ring 118a, the seat ring 118a forming a compartment when connected to the base 160a, such as by a snap-fit engagement. Referring to fig. 4C, an example of a snap engagement between a protrusion, shown as retractable protrusion 119f of impeller 118, and a groove of post 148 is shown.

The impeller 118 may include a compartment having a cover 118b for receiving one or more magnets 118c therein 118 b. The cover 118b may be removable or permanently secured in place to seal the compartment with one or more magnets.

The base 162a may also include an opening 162c, the opening 162c for receiving a portion of the post 148. This opening 162c allows the post 148 to reach the floor of the bioreactor 100 within the vessel 140. The base 160a may also include an opening 160d for allowing fluid to enter therein and be drawn into the column 148, such as when negative pressure is applied thereto, to form a drain, or to release fluid therefrom when fluid is supplied under pressure to the column 148 (such as via conduit 151).

Turning to fig. 5 and 6, a fixed bed assembly 149 can be inserted into the interior compartment of the housing 112, the assembly 149 being shown exploded in fig. 5 and assembled in fig. 6. The fixed bed assembly 149 may include a support 152 (which may be the entire fixed bed or a portion thereof, such as when portions of the fixed bed are arranged in a stacked configuration) for supporting the fixed bed 122. The support 152 can comprise a single piece or unitary structure including a central member, shown as an annular portion 154, the annular portion 154 forming a hollow annular wall corresponding to the imperforate conduit or tube 128 of FIG. 1A, with the material forming the fixed bed 122, in one example, being helically wound therearound.

The support 152 can also include an exterior, such as a support frame 156, the support frame 156 for supporting the fixed bed 122, such as from below in the arrangement shown. The support frame 156 can form an extension extending radially outward from the annular portion 154, but can allow fluid to flow therethrough, such as upward into the lower (inlet) end of the fixed bed 122. The support frame 156 can be positioned at the bottom of the annular portion 154 such that the support frame 156 provides a height at which a user can wrap or otherwise position the fixed bed 122 around the annular portion 154.

As shown, the support frame 156 includes a plurality of radially extending portions including arms 156a connected to a rim or peripheral ring 156 b. In other embodiments, the support frame 156 may include an annular shelf or disk that may include one or more apertures. In some embodiments, the support frame 156 may include a mesh or screen. In each of these embodiments, the support frame 156 may perform the dual function of supporting the fixed bed 122 while allowing fluid to flow into the fixed bed through the peripheral chamber.

The support frame 156 has an outer diameter corresponding to the inner diameter of the housing 112 such that the two structures are directly adjacent to each other and possibly in contact, but are not connected (such that the fixed bed assembly 149 can be freely inserted into and removed from the interior compartment of the housing 112). Thus, the support frame 156 can act as a locator or spacer to locate or center the fixed bed assembly 149 within the housing 112.

In another aspect, the support frame 156 can serve as a base or reference point for positioning the fixed bed 122 on the support 152. For example, if the fixed bed 122 is in the form of a sheet or fabric wrapped around the annular portion 154 during use, the support frame 156 may serve as a level against which at least one edge of the fabric may rest during the wrapping process. Specifically, if the fixed bed 122 is of the type shown in fig. 3 (or some similar material or combination of materials that may be wrapped or wrapped around a central core), the support 152 may be positioned on a surface with the support frame 156 facing downward and the fabric or other material forming the fixed bed 122 may be placed such that one edge of the material contacts the support frame 156 and the material forming the fixed bed 122 may be wrapped around the annular portion 154 using the support frame 156 as a guide.

Alternatively, if the fixed bed 122 is pre-formed or preformed before being positioned on a support 152 (e.g., an annular disk or series of disks) that is simply inserted over the annular portion 154, the support frame 156 may provide a stop or floor on which the pre-formed or preformed fixed bed 122 may rest.

The annular portion 154 may also include a separator 158, which separator 158 may divide the interior thereof into two (or more) portions 158a, 158b. The portions 158a, 158b may be the same size and shape, or may be different sizes and shapes. In any event, these portions 158a, 158b may receive, guide, and hold any tube or conduit within the interior compartment, such as conduit 151, in place for transferring fluid flow to or from the tubular post 148 associated with the container 140. In one aspect, the separator 158 may be adapted to position the tube or conduit away from the inner wall of the annular support. In this way, a tube suitable for sampling or removing a portion of the fluid within the bioreactor may be kept away from the inner wall and prevented from sucking or removing fluid from the falling film fluid flowing down the inner wall.

When forming the stack, a seal, such as an O-ring 160, may be provided for sealing the annular portion 154 from an adjacent structure (e.g., another support 152). Thus, when a plurality of fixed bed assemblies 149 are stacked within a housing, a substantially fluid impermeable or non-porous conduit or tube 128 may be formed. This may include sealing with the container 140 when the container 140 is present in the bioreactor 100. This may be achieved by a peripheral seal in the form of an O-ring 159 for sealing between the supports 152. In particular, a seal or O-ring 159 can seal the peripheral ring 156b and the inner wall of the housing 112 (see fig. 1A and 9A, which together illustrate that such an O-ring 159 is located in a recess of the ring 156b and also in contact with the upper surface of the fixed bed 122 and the inner wall of the housing 112).

When formed in a stacked form, this is optional and the fixed bed support 152 may be adapted to interlock, meaning that no welding or adhesive is required for connection for purposes of this disclosure to facilitate simple assembly and disassembly. The interlocking members may be connected in a manner that restricts at least one degree of movement of the interlocking members relative to each other. This may include limiting relative rotation between the interlocking elements and/or limiting horizontal or vertical movement between the interlocking elements. For example, this may include one or more protrusions that are received within one or more receptacles to prevent relative movement.

Specifically, as shown in fig. 6, the support 152 may include a protrusion 154a, such as the protrusion 154a on the upper end of the annular portion 154. Likewise, the support 152 may include corresponding receptacles 154b located on the underside of the annular portion 154 or elsewhere thereon. When two adjacent supports 152 are stacked, the protrusions 154a and receivers 154b thus interlock to align the parts and prevent relative rotation. It will be appreciated that the positions of the protrusion 154a and the receptacle 154b may be reversed, with a similar effect.

Returning now to fig. 4, the vessel 140, which serves as a support for the fixed bed 122 within the housing 112, may include corresponding protrusions 140d. Similar to the protrusion 154a, this protrusion 140d can interlock with the receptacle 154b of the fixed bed support 152. Thus, as shown in FIG. 1 (which may include any number), relative alignment and retention of adjacent fixed bed supports 152, as well as any additional fixed bed supports forming a stack in the housing 112, may be achieved. A seal such as an O-ring 157 may seal between the fixed bed support 152 and the vessel 140, and the vessel 140 may further include a standoff flange 140c for receiving and engaging the support (see fig. 4A).

Turning to fig. 7, 7A and 8, more details of the upper frame 130 are shown. This upper frame 130 may comprise an inner member in the form of a central ring 130a connected to an outer member, which is also in the form of a ring 130 c. The connection may include a plurality of arms 130b, and the arms 130b may extend radially between the rings 130a, 130 c. It will be appreciated that each pair of adjacent arms 130b form an opening for allowing fluid to pass through, such as from the lower outlet end of the lower fixed bed 122 associated with the support 152.

The central ring 130a may have a shape and diameter corresponding to the annular portion 154. Also, the outer ring 130c may correspond to an inner diameter of the housing 112. An O-ring 157 may be sealed between the center ring 130a and the upper end of the adjacent fixed bed support 152 (see exploded view of fig. 2).

As shown in fig. 8, 9, and 9A, the upper frame 130 may also include features to ensure proper positioning or locating, and may further provide a retaining function. For example, the outer ring 130c may also include a locating feature in the form of a receiver 130d for mating with a corresponding locating feature on the cap 114, such as a protrusion 114a (or receiver), to ensure proper alignment. The upper frame 130 may also include a receiver 130e (or protrusion) that the receiver 130e is configured to engage the protrusion 140b of the next adjacent support 152 of the fixed bed. In this way, the cover 114 secures the upper frame 130, the support 152, and the vessel 140 (also serving as a support) together in an interlocking manner, which allows for easy assembly and disassembly of the bioreactor 100.

The upper frame 130 can be sized with sufficient height/depth to hold a volume of fluid above the fixed bed 122 as the fluid exits the fixed bed 122 and before flowing into the chamber 126 formed by the non-porous conduit or tube 128, the non-porous conduit or tube 128 being formed by the annular portion 154 of the support 152. The resulting fluid well or pocket provides a stable environment for receiving the probe/sensor, which may otherwise be located in the central chamber 126 and may be either exposed to turbulence or require further insertion to access the fluid therein (which may not completely fill the respective chamber). It will be appreciated from fig. 7A that the wells or pockets formed by the upper frame 130 may be of different sizes or volumes for different uses, or may be of the same size and volume.

It can be further appreciated from a review of fig. 7A and 8 together that the number of openings formed by the arms 130b can correspond to the number of structures used to access the fixed bed 122 from the cover 114, such as not more than one structure (probe) occupying each opening. Thus, as can be appreciated from fig. 7A, multiple openings along a portion of the upper frame 130 may receive a sampler or probe 132, while other openings may not receive such a structure. To facilitate insertion and alignment during manufacture of bioreactor 100, upper frame 130 may include indicia, such as arrow 130f, to identify proper positioning of sampler/probes 132, etc., to ensure alignment with corresponding ports P in cover 114.

Another example of a bioreactor 200 is shown in fig. 10 and 10A. In this scenario, the bioreactor 200 may be similar in structure and arrangement to the larger bioreactor 100 described above, but smaller and simpler in configuration. The structural elements and terms used to describe bioreactor 200 may be the same as or overlap with those of bioreactor 100 described above, but may differ in some respects, as outlined in the description below (and such differences may be applicable to any of the disclosed embodiments).

In one example, bioreactor 200 may include a housing 212, and a fixed bed assembly 249 may be placed in housing 212, fixed bed assembly 249 including a support 252 for a fixed bed (not shown, but note a peripheral chamber 220 in which the fixed bed may be located, as further outlined below). The housing 212 may be a single piece or unitary structure such as a vessel in the form of a tub or a pot. In this case, the housing 212 may be manufactured using injection molding techniques so that no seams are present, which eliminates leakage and exposure to contamination as compared to a multi-part arrangement. In some applications, housing 212 may be translucent or transparent. In other applications, the housing 212 may be opaque and formed of any material, but inexpensive plastics are preferred, especially if made as a disposable structure.

The support 252 may define a peripheral chamber 220 between an inner surface of a wall of the housing 212 and an outer surface of the wall of the support 252. As shown, the housing 212 may be cylindrical and the support 252 may be annular. Thus, the peripheral chamber 220 may also be annular and a fixed bed located therein (which may take any form, including those described and illustrated herein).

The support 252 may include one or more locators in the form of protrusions 234. As shown, the projection 234 may include radial arms (which may be considered as tabs in a shortened form) extending toward the inner wall of the housing 212 and may engage but not be connected to the inner wall of the housing 212 to facilitate separation. Alternatively, the protrusion 234 may be part of the inner wall of the housing 212 and extend from the inner wall of the housing 212 toward the support 252, but not attached to the support 252, yet provide the desired spacing between these components.

In one aspect, the bioreactor may include an agitator, such as a magnetic impeller or stirring bar 218. This agitator may be located in a base chamber 216 formed by the lower wall 252a of the support 252 and the floor or base of the housing 212. As shown, the lower wall 252a in the interior compartment of the support 252 may include a central opening 242. This central opening 242 may fluidly connect the central chamber 226 of the support 252 with the chamber 216. Chamber 216 may further include one or more sidewalls having holes or openings that form fluid paths with peripheral chamber 220. The support 252 may include a sidewall of the chamber 216.

Thus, in operation, an agitator, such as the stirring bar 218, may be rotated via a non-contact (e.g., magnetic) coupling via an externally applied force. Agitation is used to draw fluid into the central chamber 226, through the central opening 242 and into the base chamber 216. The fluid may then be forced out through the openings therein, over or past the one or more protrusions 234, and into the peripheral chamber 220, which includes a fixed bed. Fluid exiting the peripheral chamber 220 may flow over or through the upper portion of the support 252, back into the central chamber 226, and eventually into the base chamber 216 for recirculation. One or more vanes 250 or baffles may optionally be included within the central chamber 226 to help prevent vortex formation.

Bioreactor 200 may also include a removable screw cap 214, screw cap 214 for sealing the open top of housing 112. A seal, such as an O-ring 260, may extend between the lid 214 and the housing 212. Instead of a screw cap, the cap 214 may be attached to the housing 212, such as by fasteners F (e.g., screws, bolts, clamps, clips, tabs), mating lips, or any other attachment mechanism for removably securing the cap in place, thereby allowing the bioreactor 200 to be reliably sealed from contamination during use, but easily opened.

One advantage of facilitating separation of the lid 214 and the housing 212 is that the interior of the bioreactor 200 can be easily accessed, such as before, during, or after cell culture. As a specific example, the easy-to-open lid 214 provides the advantage that the user can monitor the cell density (colony homogenization) within the fixed bed. In the particular context of DoE studies, the easy-to-open lid 214 allows, among other things, for simultaneous inoculation of multiple bioreactors 200, and then "sacrificing" individual bioreactors sequentially at a time to assess cell density (e.g., one per day) in order to establish a kinetic growth pattern of cells. This easy-to-open feature facilitates such DoE studies in a manner that a more permanent connection between the cover and the housing is not possible, and allows access to a fixed bed structure to study uniformity of cell distribution, directly into cells captured therein (even under non-sterile conditions).

The cover 214 may include one or more sensor holders 214b. These sensor holders 214b may include one or more disposable sensors for sensing parameters of the bioreactor 200. In another aspect, the sensor holder 214b may be adapted to receive a reusable sensor for sensing a parameter of the bioreactor 200.

The cover 214 may further include one or more connectors 214c, the one or more connectors 214c for connection with conduits or pipes within the support 252, such as for providing culture medium or any other additives to the bioreactor 200. Another connector 214d may be used to connect to the aspiration tube T. One or more removable caps may seal the one or more connectors. The cover 214 may optionally include one or more sampling ports 214e.

With further reference to fig. 10A, bioreactor 200 (or bioreactor 100 or any other disclosed form) may include an external temperature regulator. This regulator may take the form of a heating (or cooling) sleeve, such as blanket 262. A thermal conductor, such as a metal (e.g., aluminum) vessel, may facilitate heat exchange between the regulator and the bioreactor 200. In one aspect, the blanket 262 and vessel 264 are not part of the disposable bioreactor 200, but are part of a system.

Referring to fig. 11, 12, 13, 14 and 15 and their associated parts, as outlined below, another example of a bioreactor 300 is shown. As in the above example, bioreactor 300 may be similar in structure and arrangement to bioreactor 100 and/or bioreactor 200 described above. The structural elements and terms used to describe bioreactor 300 may be the same as or overlap with the structural elements, terms, and reference numerals of bioreactor 100 and/or 200 described above, but may differ in some aspects as outlined in the description below (and such differences may apply to any of the disclosed embodiments).

Bioreactor 300 may include a housing 312 with a support 352. In one example, the housing 312 may be a unitary, one-piece, or monolithic container, such as a canister, a barrel, or a canister. In such cases, the housing 312 may be manufactured using injection molding techniques to eliminate seams and attendant leakage or contamination. In some applications, the housing 312 may be translucent or transparent. In other applications, the housing 312 may be opaque and made of any material, preferably plastic, if disposable is desired.

Referring to fig. 11A and 11B, the support 352 may define a peripheral chamber 320 between an inner surface of the housing 312 and an outer surface of the support 352. As shown, the housing 312 may be circular in cross-section and the support 352 may be annular in shape. Thus, the peripheral chamber 320 may also be annular. The peripheral chamber 320 may be adapted to receive a fixed bed 322. The fixed bed 322 may be any type of fixed bed mentioned hereinabove.

As can be appreciated from fig. 11 and fig. 12 and 12A, one or more locators, such as protrusions or arms 346, may position the support 352 within the housing 312. The arms 346 may extend radially outward from a lower portion of the support 352. In one aspect, three such arms 346 are substantially equally spaced from each other about the circumference of the support 352. These arms 346 may serve to center the support 352 and maintain a fixed distance between the support 352 and the wall of the housing 312, thereby at least partially defining the size of the peripheral chamber 320. On the other hand, the arms 346 may support the fixed bed 322 from below (shown transparent in fig. 11 for clarity) while still allowing fluid to flow through the peripheral chamber and the fixed bed 322 therein, which may be best seen in fig. 11 and 15A.

The housing 312 may include a receiver 348 adapted to engage the at least one arm 346. The receiver may be positioned at a lower portion of the housing 312 so as to correspond to the position of the arms 346 of the support 352. The receiver 348 may be in the form of a ledge, shelf, bracket, pocket, slot, recess, or other element adapted to receive and/or cradle and support one or more of the arms 346, and thus the support 352, within the body of the housing 312. The receptacle 348 may also take the form of a circumferential lip extending along the inner surface of the housing 312. Alternatively, the positions of the arms 346 and the receivers 348 may be reversed such that the arms are connected to the housing 312 and the receivers 348 are connected to the support 352, but remain removably attached to each other.

In one aspect as shown in fig. 12A, the receiver 348 may include a plurality of individual receivers, such as a plurality of shelves or ledges, spaced apart from each other and adapted to engage the plurality of arms 346. The receiver 348 may include one or more stops 349, the stops 349 being adapted to engage the arms 346 and, once engaged, limit rotation of the arms 346. For example, the stop 349 may include one or more sidewalls associated with the receiver 348. Thus, the receiver 348 may form a pocket or recess adapted to receive the arm 346 and prevent rotation of the support 352 within the housing 312. In this case, the arms 346 may be used to: (1) Maintaining a space between the support 352 and the walls of the housing 312 (e.g., maintaining a constant size of the fluid chamber 320); (2) Maintaining the support 352 at a desired height within the housing 312; and/or (3) prevent rotational movement of support 352 within housing 312.

In another aspect, the locator or arm 346 can serve as a base or reference point for locating the fixed bed 322 on the support 352. For example, if the fixed bed 322 is in the form of a sheet or fabric wrapped around the support 352 during use, the arms 346 may serve as a level against which at least one edge of the fabric may rest during the wrapping process. Specifically, if the fixed bed 322 is of the type shown in fig. 3 (or some similar material or combination of materials that may be wrapped or wrapped around a central core), the support 352 may be positioned on a surface with the arms 346 oriented downward and the remainder of the support 352 oriented upward, and the fabric or other material forming the fixed bed 322 may be placed such that one edge of the material contacts the arms 346 and wrapped around the annular portion of the support 352 using the arms 346 as guides, the material forming the fixed bed 322 may be wrapped around the annular portion of the support 352.

Alternatively, if the fixed bed 322 is pre-formed or preformed prior to being positioned on a support 352 (e.g., an annular disk or series of disks) that is simply inserted over an annular portion of the support 352, the arms 346 may provide a stop or floor upon which the pre-formed or preformed fixed bed 322 may rest.

In one aspect, bioreactor 300 may include an agitator, such as magnetic impeller 318. The magnetic impeller 318 may be in the form of a "plug-in" rotatable non-contact magnetic impeller 318, which thus forms a centrifugal pump in the bioreactor. In one aspect, the impeller 318 may include one or more magnet caps that may be attached (e.g., glued) to ensure that the magnets are sealed to the impeller. The agitator may also be in the form of an impeller mechanically coupled to the base, an external pump forming part of the fluid circulation system, or any other means for inducing fluid circulation within the bioreactor. As shown, the impeller 318 may include a body including curved walls or vanes adapted to draw fluid therein and expel fluid radially outwardly therefrom.

A magnetic impeller 318 may be positioned in the base chamber 316. In one aspect, this base chamber 316 may be defined at least in part by a lower wall of the support 352 and a lower wall of the housing 312. This base chamber 316 may be in fluid communication with an outer chamber 320. In one aspect, the arms 346 extend between the base chamber 316 and the outer chamber 320.

The retainer holds the impeller 318 in place, in the example shown, in the form of a post 370. This post 370 may be adapted to support and maintain the position of the impeller 318 within the base chamber 316. The post 370 may include a base 372 adapted to support at least a portion of the impeller 318 from below. The post 370 may include a race that may be adapted to receive a bearing 374 to facilitate rotation of the impeller 318. The bearing 374 may include one or more ball bearings, cylindrical bearings, bushing material, or any other bearing element suitable for facilitating relative movement between the impeller 318 and the post 370. In one aspect, the impeller 318 may include a race or portion of a race for the bearing 374, as shown in fig. 13 (see also bioreactor 200 in fig. 4B, but may be similarly applied to bioreactor 300). In one aspect of the present disclosure, the column 370 may be tubular or hollow, and thus may be used, in particular, as a conduit for introducing fluid into the bioreactor 300 or withdrawing fluid from the bioreactor 300 via the chamber 316.

As can be seen in fig. 13, the support 352 may include a central opening 342. This central opening 342 may connect the central chamber 326 of the support 352 with the base chamber 316. The post may be adapted to pass through the central opening 342 of the support 352 to position and center the impeller 318 relative to the support 352.

The post 370 may include a stop 378, the stop 378 being configured to limit the insertion distance through the central opening 342. For example, this stop 378 may include a shoulder, lip, detent, protrusion, or other element that may engage the support 352 and prevent the post 370 from passing further through the central opening 342. As shown, the stop 378 includes a first portion of the post 370 that has a diameter greater than the diameter of the central opening 342. The second portion of the post 370 may have a diameter that is smaller than the diameter of the central opening 342, and thus may pass through the central opening 342 and into the central chamber 326.

The stop 378 may be located a distance from the base 372 such that the height of the impeller 318 may fit therebetween. In one example, as shown, the distance from the base 372 to the stop 378 is sufficiently greater than the height of the impeller 318 to suspend the impeller 318 therebetween, with at least some clearance between the impeller and the bottom wall of the support 352. In one embodiment, when the bioreactor 300 is fully assembled, the posts 370 extend below the bottom of the impeller 318 to suspend the impeller 318 above the floor of the housing 312.

Referring to fig. 12A and 13, the lower wall of the support 352 may include one or more fluid openings 380. Each fluid opening 380 may be adapted to transfer fluid from the central chamber 326 to the impeller chamber 316 for distribution to the fluid chamber 320. The fluid openings 380 may be equally spaced about the central opening 342. As shown, bioreactor 300 includes three fluid openings separated from one another by separator 382, but any number of fluid openings may be present and any means of providing such openings in support 352 may be utilized as long as uniform fluid flow is facilitated. The separator 382 may extend from a peripheral portion of the support 352 to a central body 384 of the support 352, the central body 384 may receive the post 370.

In operation, the impeller 318 may rotate, drawing fluid downward through the central chamber 326, through the fluid opening 380, and into the chamber 316. The fluid may then be forced outwardly and upwardly into and through the peripheral chamber 320 between the support 352 and the housing 312. After the fluid rises through the peripheral chamber 320 (which may include the fixed bed 322), the fluid may pass over or through the upper portion of the support 352 and back into the central chamber 326 for recirculation. Although not shown in this embodiment of the bioreactor, one or more vanes or baffles may prevent the fluid from forming a vortex during recirculation (see, e.g., vanes 260 of bioreactor 200).

The support 352 may be of unitary construction, such as an injection molded construction. As will be appreciated from the foregoing, this single piece support 352 may serve as a pump housing or vessel, a fixed bed support, an anti-vortex structure, forming a central chamber for receiving fluid (such as a falling film from chamber 220), and preventing all associated components from moving within housing 312.

Returning to fig. 11 and 12, and referring to fig. 15, housing 312 may include one or more ports 360 to allow measurement of characteristics of bioreactor 300. For example, port 360 may be adapted to receive or hold a sensor adapted to collect information and/or measure a characteristic of the bioreactor (e.g., any of pH, dissolved Oxygen (DO), temperature, cell density, flow rate, metabolite level, medium or medium composition level, or other parameter, but is not limited thereto). In one aspect, the sensor may be an optical sensor.

In one embodiment, as shown in fig. 14, ports 360 may be located in communication with the peripheral chamber 320. More specifically, the ports 360 may be located in an upper portion of the peripheral chamber 320. Even more specifically, the ports 360 may be located in the peripheral chamber 320 at a position above the height of the fixed bed 322. Having a port 360 at this location places the port and sensor at a location in the fluid that has passed through the fixed bed 322 under normal flow conditions.

The port 360 may include a support section, such as a tube or a shelf, to support at least a portion of the weight of the sensor. In one aspect, the port 360 may include a cylindrical bore that allows the sensor to directly access the interior of the bioreactor 300. In another aspect, port 360 may be sealed from the interior of the bioreactor, but may allow a non-contact sensor, such as an optical sensor, to sense a parameter inside the bioreactor. Port 360 may include a lock for securing the sensor in place relative to the port. The lock may maintain a seal between the interior of bioreactor 300 and the external environment. For example, port 360 may include a luer lock or other component for maintaining sterility of bioreactor 300 while allowing a sensor to sense a parameter inside bioreactor 300.

Bioreactor 300 may further include a cover 314 for sealing bioreactor 300. As shown in fig. 14, the cover 314 may be removably secured to the housing 312, such as by a threaded connection. Alternatively, cover 314 may be attached to housing 312 by one or more fasteners, such as screws, bolts, clamps, clips, tabs, mating lips, or any other attachment mechanism that allows bioreactor 300 to be easily opened. The housing 312 and the cover 314 may together comprise a container for housing the remaining elements of the bioreactor.

One advantage of using an easy-open connection between the lid and the housing is that the interior of the bioreactor 300 can be easily accessed, such as during or after an experimental run. For example, in the case of fixed bed bioreactors, an easily openable lid provides the advantage of potentially helping to monitor cell density (colony homogenization) within the fixed bed by direct measurement. In the case of DoE studies, the easy-to-open lid allows, among other things, simultaneous inoculation of multiple bioreactors 300, followed by successive "sacrifice" of a single bioreactor at a time (e.g., one per day) in order to establish a dynamic growth pattern of cells over time. This easy-to-open feature facilitates such DoE studies in a manner that a more permanent connection between the lid and the housing is not possible.

A seal such as an O-ring 315 may form a fluid tight connection between the cap 314 and the housing 312. In some embodiments, the O-ring 315 and/or a portion of the cover 314 or housing 312 adapted to contact the O-ring 315 may be crimped to facilitate clamping between the elements. In some aspects, the O-ring 315 and/or a portion of the cap 314 or housing 312 adapted to contact the O-ring 315 may be smooth to facilitate proper sealing between the components.

Referring to fig. 11 and 11A, the cover 314 may include one or more connectors or ports P. These ports P may be adapted to connect to one or more conduits, such as for adding or removing culture medium or any other additives or products or waste to or from the bioreactor 300. One or more removable covers may be provided, in some cases including suitable seals for sealing one or more ports. These ports may include one or more of a first port 390a for adding medium, a second port 390b for removing medium, a third port 390c for adding gas, a fourth port 390d for removing gas, a fifth port 390e for inserting or holding a probe (e.g., a temperature probe), and a sixth port 390f for adding a pH adjuster (e.g., a base).

In some embodiments, port 392 may allow a user to empty bioreactor 300. As shown in fig. 11B and 14, the port 392 may be connected to a conduit 394. In one embodiment, the conduit 394 may be fluidly connected to the column 370. In such embodiments, the post 370 may include a tube or other hollow space to which the conduit 394 may be connected. The column 370 may include a base 372 with an opening to allow for emptying of the bioreactor 300, or if operated in reverse, this may instead introduce fluid therein.

The conduit 394 may also be used to maintain the relative position between the various portions of the bioreactor 300. Specifically, the conduit 394 extends along the post 370 to prevent it from exiting through the central opening 342 in the support 352. For example, referring to fig. 14, the lower end of the conduit 394 may be placed along the post 370, such as at the point of contact between the supports 352 around the central opening 342. In some embodiments, a connector or other support (not shown) may connect the conduit 394 to the post 370, such as a collar. In other embodiments, the conduit 394 is sized such that an interference or friction fit prevents it from breaking. Thus, the conduit 394 may trap or sandwich the bottom wall of the support 352 between the conduit 394 and the post stop 378. This maintains the relative positions of the conduit 394, the post 370 and the impeller 318 so that these elements do not become disconnected after assembly.

To facilitate assembly of the bioreactor 300, the length of the conduit 394 may be greater than the height of the bioreactor 300, or greater than the distance between the bottom of the support 352 and the cover 314 (in the assembled state). For example, the conduit 394 may be 10% to 60% longer than the height of the bioreactor 300. In one embodiment, the bioreactor 300 may be 50 millimeters high and the conduit 394 may have a length between 55 millimeters and 80 millimeters. In this way, the post 370 may pass through the impeller 318 with the bearing 374 therebetween. The post 370 may extend through the central opening 342 of the support 352, such as until the stop 378 contacts the underside of the support 352. The conduit 394 may be connected from the top to the post 370 such that it prevents vertical movement relative to the support 352.

It will be appreciated that the assembly comprising the conduit 394, support 352, post 370, bearing 374 and impeller 318 may be inserted into the housing 312 as a single unit. This insertion may be performed simultaneously with or prior to insertion of the fixed bed 322 into the housing 312. Once inserted into the housing 312, the other end of the conduit 394 may be connected to the port 392 and the cover 314 may be attached to the housing 312.

In other aspects, once the conduit 394, support 352, post 370, bearing 374, and impeller 318 are inserted into the housing 312, any additional length of the conduit 394 may extend through a hole or aperture in the cover 314 where the port 392 is located. The extra length of the conduit 394 may be cut to length, such as within about 10 millimeters above the level of the cover 314. The free end of the conduit 394 may be attached to the port 392, and then the port 392 may be pushed down into the hole or aperture and secured to the cap 314, such as with glue (e.g., UV glue), welding, or other fastening means.

Referring to fig. 15A and 15B, bioreactor 300 may be adapted to maintain the position of support 352 within bioreactor 300. As shown, an overhang portion of the lid 314, such as the vertical leg 398, may extend downward from the lid 314. The legs 398 may interlock with the support 352 or be adjacent to the support 352. The legs 398 may also have a length corresponding to the distance from the lid 314 to the top of the annular wall of the support 352, particularly when the lid is fully positioned on the housing 312 due to the threaded connection used in the illustrated embodiment.

Thus, when bioreactor 300 is assembled with lid 314 in place over the open top of housing 312, legs 398 are adjacent to and may even contact supports 352. In either case, this prevents the carrier from moving vertically any appreciable distance or floating within the bioreactor 300. As shown, the leg 398 may include a substantially flat lower edge for engaging the support 352. However, the legs 398 may include one or more extensions, fingers, clamps, clips, supports, or any other structure for engaging the support 352 and preventing it from rising vertically within the bioreactor 300. Legs 398 may also be connected to support 352 instead of engaging cover 314.

In other embodiments, an upper frame such as shown in fig. 8 and described above may be positioned between the cover 314 and the support 352 in place of or in addition to the legs 398 to maintain the position of the support 352 within the housing.

As described in the previous embodiments, the bioreactor 300 may be equipped with a temperature controller, such as a heating (or cooling) blanket and/or thermal conductor as described herein. Similarly, bioreactor 300 may be used for similar purposes in the same manner as described herein with respect to bioreactor 200. In another aspect, bioreactor 300 may be similar in configuration to bioreactor 100 described previously, but on a smaller scale. The more similar the configuration of the bioreactor 300 and the larger bioreactor 100, the more likely the optimized or determined parameters in the small scale bioreactor will lead to similar results for the process described in the larger bioreactor 100.

Fig. 16 illustrates in flow chart form exemplary steps of assembling bioreactor 100 according to a first embodiment of the present disclosure. If not preassembled, the method may include a step 400 of installing a magnet in the body of the impeller 118. Step 402 may include coupling the post 148 and bearings to the impeller 318 and the vessel 140. A next step 404 may include placing an O-ring on the container 140 and positioning it in the housing 112. If present, the conduit 151 is connected to the container 140 at step 406.

At step 408, the method may further include forming a fixed bed assembly 149, which in the case of a spiral or coiled form, the fixed bed assembly 149 may include rolling the fixed bed material onto the support 152 and placing the O-ring 157 on the vessel 140. At step 410, a fixed bed assembly 149 may be placed in the housing 112 to interlock with the vessel 140. If multiple stacks are desired, at step 414, an O-ring 159 may be positioned over the fixed bed 122 of the first fixed bed assembly and steps 408, 410, 412 may be repeated as desired.

At step 414, an o-ring 157 is associated with the uppermost fixed bed assembly 149 in the stack to form a seal and interlocked with this fixed bed assembly 149 at the upper frame 130. At step 416, a sampler or probe may be inserted into the uppermost fixed bed assembly 149 through the upper frame 130 in a desired orientation. If present, the conduit 151 may be connected to the cover 114, and the cover 114 may be interlocked with the upper frame 130 and sealed to the upper frame 130, as indicated at step 418.

Similar steps may be performed for bioreactors 200 and 300, varying as desired according to different arrangements (e.g., O-rings and repeated assembly steps may be omitted since only one fixed bed is provided in the illustrated embodiment of bioreactors 200, 300). In any of the methods disclosed, the steps may be performed or carried out by a different person or a care-giver, and may be carried out in a different order than shown. Furthermore, not all steps have to be performed and the order of the steps may be different.

In another embodiment, referring to fig. 17, it may be advantageous to simultaneously provide a system S comprising a plurality of bioreactors 500, such as any of the bioreactors disclosed herein. In one aspect, a single controller may control various aspects of each of the plurality of bioreactors. Such simultaneous control may allow for the efficiency of operation or application of one or more different parameters of a given process in order to optimize the conditions of the process. For example, such simultaneous control may allow for control of different cell culture conditions in different bioreactors, including but not limited to pH, DO, temperature, agitation rate, flow rate, and the like.

System S may allow for testing and/or optimization of operating parameters for a given process within bioreactor 500. Once the desired operating parameters are determined, it may be desirable to scale up the process from small-scale bioreactor 200 to a bioreactor capable of running an optimized or desired process at the determined parameters. Thus, in another aspect of the present disclosure, the small-scale bioreactors 200, 300 may be similar in configuration to the large-scale bioreactors 100. The more similar the configuration of the small-scale bioreactors 200, 300 and the larger bioreactor 100, the more likely the optimized or determined parameters in the small-scale bioreactors will lead to similar results for the process described in the larger bioreactor 100.

Bioreactor 500 is connected to a single controller 502 forming part of a system S, which is shown with eight bioreactors, but may include any number of more than one. Controller 502 may include a computer, microprocessor, mobile device, or any other control device suitable for monitoring and/or regulating conditions within bioreactor 500. In one aspect, there may be one or more sensors for monitoring one or more environmental conditions or parameters within a given bioreactor. One or more multiplexers may allow signals (e.g., sensor and/or control signals) to be transmitted through a single connector (e.g., a wire). This may reduce the number of components within the system required to monitor an environmental condition or parameter (or transmit/receive control signals).

The system S may be modular such that one or more bioreactors 500 are interconnected to form the system. For example, the system may include a single controller 502 connected to two bioreactors 500. The two bioreactors may operate separately or may be a pair of connected bioreactors and may include additional single bioreactors. In one aspect, one or more additional pairs of bioreactors may be included. The manifold M may be connected to a plurality of bioreactors 200. The embodiment shown in fig. 16 shows four pairs of bioreactors 500, but more or fewer bioreactors or bioreactor pairs may be included in the system.

The controller 502 may be adapted to monitor and/or control various process parameters of each of the bioreactors 500. For example, the controller 502 may be adapted to control one or more of temperature, pH, DO, agitation speed, or flow rate in a given bioreactor 500. In one aspect, the controller 502 may be adapted to individually control one or more of the parameters in at least one bioreactor, wherein the parameters are different from the same parameters in another bioreactor. In another aspect, the controller 502 may be adapted to individually control all parameters in each of the plurality of bioreactors 500 in the system. Thus, a single controller 502 may be adapted to control different parameter sets in each bioreactor 500. This may allow multiple parallel process conditions to run simultaneously. Such parallel process runs may allow multiple parameters to be tested simultaneously, such as condition optimization for a given process. One advantage of such parallel process runs is that parallel bioreactors can be operated using different DoE (design of experiment) parameters and can guide the user to scale up a given culture to a larger bioreactor and/or to obtain data in an efficient manner.

Running multiple processes simultaneously can save time, but may be inefficient if done on a large scale. Thus, in another aspect of the present disclosure, each bioreactor 500 may comprise a small volume bioreactor, such as a bioreactor having a volume of 300 milliliters or less. In some examples, the bioreactor can have any operating volume of 250 milliliters, 200 milliliters, 150 milliliters, 100 milliliters, 80 milliliters, 50 milliliters, or less. The use of such a smaller volume bioreactor 500 in system S may allow for efficient use of laboratory space and consumable resources, such as culture medium. Furthermore, a smaller bioreactor 500 like this is necessarily smaller in size than a larger reactor, and thus cheaper to manufacture, and thus more economical to use than a larger suspension reactor. Thus, the use of multiple smaller bioreactors 500 may facilitate efficient process development or DoE research.

In another aspect, bioreactor 500 may be a disposable bioreactor that may include probes or other sensors for measuring parameters thereof. In one aspect, such probes or other sensors may be discarded with bioreactor 500. In another aspect, bioreactor 500 may be adapted to receive reusable probes or sensors in a sterile manner. The probe or sensor may include one or more sensors for sensing cell density, optical density, pH, DO, temperature, agitation rate, flow rate (including flow into, out of, and within the bioreactor, such as during mixing), or other desired parameters associated with cell culture.

In another aspect, the use of small scale bioreactors allows for the development and testing of processes that can be effectively scaled up to larger reactors. Conversely, a small scale bioreactor may facilitate scaling down a process from a larger bioreactor to more efficiently understand or characterize the process. This ability to scale up and/or down may be particularly relevant in the field of viral vectors, as the developer and producer work together closely to build up large production capacities in a shorter time.

To allow for enlargement and/or reduction between reactor sizes, a smaller bioreactor should be structurally and/or functionally representative of a larger bioreactor. In the case of a fixed bed reactor, the smaller bioreactor may have the same fixed bed height and/or compactness as the larger bioreactor. In the case of fixed beds which are highly similar, the diameter of the fixed bed in a smaller bioreactor can be smaller to achieve a lower surface area. One way to allow expansion in fixed bed reactors is to maintain such structural similarity, as well as similar oxidation and mixing strategies, which may allow direct scale up/down of the process between different sized reactors.

Examples：

In a non-limiting example illustrating the expansion between a smaller bioreactor and a larger bioreactor, two test runs performed in the smaller bioreactor are used to compare the results with the known results for the larger bioreactor. The bioreactor conditions used are listed in table 1.

Table 1: bioreactor culture conditions

As a first step, adherent HEK293 cells from the cryopreserved cell bank were thawed. Cells were pre-cultured in T-flasks and passaged every 3 to 4 days (seeded at 20,000 cells/cm and harvested at mid-exponential phase) using DMEM medium enriched with 5% bovine serum and 1% antibiotic-antifungal agent (4.5 g/l glucose/l).

The cells were further expanded in a cell factory (Nunc) before seeding the mini-bioreactor. Cells were then seeded at 20,000 cells per square centimeter in a smaller bioreactor (e.g., scale-X of the present inventors) with a volume of 0.5 square meters ^TM Bioreactor) and maintained in batch mode for 2 to 4 hours to adapt to their new environment, then the growth phase was started using a recirculation loop (0.17 ml/cm) containing growth medium-DMEM (4.5 g/l glucose) enriched with 5% bovine serum. Supernatant samples were taken daily to evaluate glucose and lactate distribution. Harvesting was performed at the end of the run to assess cell density in the fixed bed.

The bioreactor was emptied and rinsed with DPBS solution containing 5 mm EDTA prior to retrieving the fixed bed for cell counting. The fixed bed is then removed from the bioreactor. Cell density and homogeneity were estimated at different locations of the fixed bed (N, S, W, E, inside and outside at the top and bottom, samples were taken with 1 square centimeter of each surface area; see FIG. 18).

After each test run, the density and viability of the suspended cells in the fixed bed samples were measured by Trypan Blue (Trypan Blue) dye exclusion method using a Thoma hemocytometer. Biomass estimation of 1 cm square PET samples was performed by acid cell lysis followed by crystal violet staining and nuclear counting. The average cell densities achieved for runs 1 and 2 were 617,032 cells per square centimeter and 673,932 cells per square centimeter, respectively, and the results are summarized in table 2.

Table 2: the key value of bioreactor culture; final cell density and population doubling time after disassembly (PDT)

The uniformity of cell distribution was demonstrated by two runs of the mini-bioreactor by cell counting at different locations on a fixed bed. The results of these cell counts are shown in table 3 below. At the end of the culture, the maximum difference observed was lower compared to the cell density over the whole stationary bed, only slightly exceeding 15% at one location.

Table 3: cell count at different locations on a fixed bed of a mini-bioreactor

Furthermore, as shown in fig. 19, the pH and Dissolved Oxygen (DO) trends maintained good stability throughout the run of the mini-bioreactor. During the phase opportunity (e.g., equilibration and inoculation), the pH and DO will experience the expected peak or change, but overall, the levels remain significantly stable throughout the run. The pH value is kept stable between 7.2 and 7.4, and completely accords with the set value. DO never reached below 100%, indicating that the cell growth oxygenation limit of the bioreactor was never approached.

In addition, various metabolites measured in the mini-bioreactor during the pilot run were compared to metabolites present during similar runs in the larger reactor (e.g., scale-X Hydro (2.4 square meters), the carbon (10 square meters) product of the present inventors). As shown in fig. 20, in similar time runs, the glucose consumption trend was similar between different reactors. In addition, the yields of lactic acid in the different reactors also show a similar trend.

The small scale bioreactor 500 is a successful proof of concept showing that the fixed bed configuration is easily retrofitted to produce a small scale commercial bioreactor of 0.5 square meters. Tests have shown that the same performance as the larger bioreactors in the series can be reliably achieved in smaller scale protocols. Cell growth, cell distribution and metabolite behavior data are equivalent to the same process run in larger scale bioreactors. The new small scale system that proved to remain straightforward to expand into more efficient and low cost process development, process optimization, and scaling down research opens up possibilities. While the presently contemplated bioreactors range from 2.4 square meters to 600 square meters of growth surface, this study offers the prospect of an extended range with a minimum size that is about 5 times lower than larger models, reducing the development costs, operations, and time associated with operating smaller bioreactors.

Summarizing the various aspects that may be involved in the present disclosure, the following examples are determined, which may be arranged in any combination:

1. an apparatus for culturing cells, comprising:

a bioreactor comprising a housing having walls forming an interior compartment;

a fixed bed for culturing cells;

a fixed bed support for a fixed bed, the fixed bed support adapted to be removably positioned within the interior compartment of the housing; and

one or more locators for uniformly spacing the fixed bed support from the wall of the housing.

2. The apparatus of example 1, wherein the fixed bed support comprises a central portion and a peripheral portion, the peripheral portion extending radially outward from the central portion and comprising one or more locators, the peripheral portion having an outer diameter corresponding to an inner diameter of the housing.

3. The apparatus of example 2, wherein the peripheral portion comprises one or more protrusions.

4. The apparatus of example 2 or example 3, wherein the peripheral portion comprises an outer ring connected to the central portion by one or more protrusions.

5. The apparatus of any one of examples 2-4, wherein the peripheral portion comprises an annular disc-shaped surface having a plurality of apertures therein.

6. The apparatus of any one of examples 2 to 5, wherein the peripheral portion comprises a mesh or screen.

7. The apparatus according to any one of examples 2 to 6, wherein the peripheral portion is adapted to support the fixed bed from below.

8. The apparatus of any one of examples 2-7, wherein the peripheral portion is adapted to allow fluid to flow therethrough.

9. The apparatus of any one of examples 2 to 8, wherein the central portion includes a separator that forms a plurality of spaces in an interior of the central portion for receiving the conduit within the housing.

10. The apparatus of example 9, wherein the plurality of spaces have different sizes.

11. The apparatus of any one of examples 9 to 10, wherein at least one of the conduits is adapted to convey the fluid flow to or from an interior of the central portion.

12. The apparatus of example 11, wherein the separator is adapted to position at least one of the conduits, the conduits being adapted to convey a fluid flow away from an inner wall of the central portion to prevent contact between the conduits and falling film fluid flowing down the inner wall.

13. The apparatus of any one of examples 2 to 12, wherein the central portion of the support forms a receptacle for receiving the agitator.

14. The apparatus of example 13, wherein the agitator comprises an impeller adapted to rotate about an agitator support, the agitator support adapted to receive and retain the fixed bed support.

15. The apparatus of example 14, wherein the agitator support comprises a tubular post connected to a flexible drain connected to a lid of the bioreactor.

16. The apparatus of example 15, wherein the peripheral portion of the fixed bed support comprises one or more protrusions in the form of a plurality of radially extending arms.

17. The apparatus of example 16, wherein the plurality of radially extending arms are connected to a rim, the rim having an outer diameter corresponding to an inner diameter of the housing.

18. The apparatus of example 16, wherein the plurality of radially extending arms engage and support the fixed bed.

19. The apparatus of any of examples 16-18, wherein the housing includes a receiver for receiving at least one of the one or more protrusions.

20. The apparatus of any of examples 2 to 19, wherein the central portion and the peripheral portion of the fixed bed support comprise a single unitary structure.

21. The apparatus of any one of examples 1 to 20, wherein the fixed bed comprises one or more layers of woven or nonwoven material wrapped around a central portion of the fixed bed support.

22. The apparatus of any one of examples 1 to 21, further comprising a seal for sealing between the inner wall of the housing and the fixed bed support.

23. The apparatus of any one of examples 1 to 22, wherein the fixed bed comprises a plurality of fixed bed sections, the fixed bed support further comprising a plurality of interlocking support sections for supporting each of the plurality of fixed bed sections.

24. The apparatus of example 23, wherein each interlocking support portion is adapted to interlock with an adjacent support portion.

25. The apparatus of example 23 or example 24, further comprising a first seal for sealing each adjacent interlocking support member portion together.

26. The apparatus of example 25, further comprising a second seal for sealing each of the plurality of portions to an inner wall of the housing.

27. The apparatus of any one of examples 1 to 26, further comprising an upper frame for positioning above the fixed bed.

28. The apparatus of example 27, wherein the upper frame has a height sufficient to form at least one pocket for allowing fluid to accumulate therein when exiting an upper end of the fixed bed, and a sensor for sensing a characteristic of the fluid in the at least one pocket.

29. The apparatus of example 27 or example 28, wherein the upper frame is adapted to engage a cover of the bioreactor.

30. The apparatus of any one of examples 27-29, wherein the upper frame forms a plurality of pockets, the plurality of pockets having different volumes.

31. The apparatus of any of examples 27 to 30, wherein the upper frame is adapted to receive or align one or more samplers for sampling the fixed bed.

32. The apparatus of any one of examples 1 to 31, further comprising a cover removably coupled to the housing.

33. The apparatus of example 32, wherein the cover is adapted to hold the fixed bed vertically in place within the housing.

34. The apparatus of example 32 or example 33, wherein the cover comprises an overhang portion for engaging the fixed bed or fixed bed support.

35. The apparatus of any of examples 32-34, wherein the cap is adapted for threaded engagement with the housing.

36. The apparatus of any one of examples 32-35, further comprising a gasket between the cover and the housing.

37. The apparatus of any one of examples 1 to 36, further comprising a port in a wall of the housing, the port being above an upper end of the fixed bed when positioned therein.

38. The apparatus of any one of examples 1-37, wherein the housing comprises a one-piece rigid structure forming the interior compartment.

39. An apparatus for culturing cells, comprising:

a housing and a lid together defining a container having an interior compartment; and

an assembly for positioning within the interior compartment, the assembly comprising a fixed bed adapted to culture cells, wherein the assembly is adapted to interlock with the container to maintain the position of the fixed bed within the interior compartment.

40. The apparatus of example 39, wherein the assembly comprises an upper portion adapted to interlock with the lid.

41. The apparatus of example 40, wherein the upper portion includes an upper frame for positioning above the fixed bed.

42. The apparatus of example 41, wherein the upper frame has a sufficient height to form at least one pocket for allowing fluid to accumulate therein when exiting an upper end of the fixed bed, and a sensor for sensing a characteristic of the fluid in the at least one pocket.

43. The apparatus of example 41 or example 42, wherein the upper frame includes a recess for engaging a protrusion extending from a lid of the bioreactor.

44. The apparatus of any one of examples 41-43, wherein the upper frame forms a plurality of pockets, each having a different volume.

45. The apparatus of any one of examples 41-44, wherein the fixed bed comprises a plurality of fixed bed portions, each fixed bed portion associated with one of a plurality of supports adapted to interlock with an adjacent support.

46. The apparatus of any one of examples 41-45, wherein the upper frame is adapted to interlock with at least one of the plurality of supports.

47. The apparatus of any of examples 45-46, wherein each of the plurality of supports comprises a central portion and a peripheral portion, the peripheral portion adapted to allow fluid to reach the fixed bed, the peripheral portion having an outer diameter corresponding to an inner diameter of the housing.

48. The apparatus of any of examples 45 to 47, further comprising at least one O-ring located between at least two of the plurality of supports or between the cover and at least one of the plurality of supports.

49. The apparatus of any of examples 45-48, wherein the cover is adapted to provide downward pressure on the assembly when the cover is connected to the housing.

50. The apparatus of example 49, wherein the downward pressure is sufficient to maintain the at least one O-ring in place without glue.

51. The apparatus of any one of examples 39 to 50, wherein the assembly comprises a lower portion for receiving the agitator, the lower portion adapted to interlock with at least one support for supporting the fixed bed.

52. An apparatus for culturing cells, comprising:

a bioreactor comprising a housing having walls forming an interior compartment;

a fixed bed for culturing cells; and

a support for a fixed bed, the support being adapted to be removably positioned within the interior compartment, the support at least partially forming a chamber housing the agitator and including one or more centering projections extending toward a wall of the housing.

53. The apparatus of example 52, wherein the protrusions engage the fixed bed.

54. The apparatus of example 52 or example 53, wherein the housing includes one or more receptacles for receiving the one or more protrusions.

55. The apparatus of any of examples 52 to 54, wherein the engagement between the one or more protrusions and the one or more receptacles is adapted to prevent rotation of the support within the housing.

56. An apparatus for culturing cells, comprising:

a housing having walls forming an interior compartment;

a fixed bed for culturing cells within the interior compartment;

One or more probes for extending into the interior compartment, adjacent to or into the fixed bed; and

an upper frame covering the fixed bed for holding the fixed bed and organizing the one or more probes.

57. The apparatus of example 56, wherein the upper frame includes one or more indicia to indicate a position or orientation of the one or more probes.

58. The apparatus of example 56 or example 57, wherein the one or more probes are attached to the upper frame.

59. An apparatus for culturing cells, comprising:

a one-piece housing;

at least one fixed bed for culturing cells; and

a plurality of fixed bed supports adapted to interlock for positioning within a single piece housing.

60. The apparatus of example 59, wherein each of the plurality of fixed bed supports comprises an annular portion and a support frame extending radially outward from the annular portion.

61. The apparatus of example 60, wherein the support frame has an outer diameter corresponding to an inner diameter of the housing.

62. The apparatus of example 60 or example 61, wherein the support frame comprises a substantially planar extension.

63. The apparatus of any of examples 60 to 62, wherein the support frame is adapted to allow fluid to flow therethrough.

64. The apparatus of any one of examples 60-63, wherein the support frame extends from a bottom of the annular portion.

65. The apparatus of any one of examples 60 to 64, wherein the support frame comprises a plurality of radially extending arms connected to the peripheral ring.

66. The apparatus of any one of examples 60 to 65, wherein the support frame comprises a mesh or screen.

67. The apparatus of any one of examples 60 to 66, wherein the support frame is adapted to support the at least one fixed bed from below.

68. The apparatus of any of examples 60 to 67, wherein the support frame is adapted to serve as a base to position the fixed bed on a fixed bed support.

69. The apparatus of any of examples 59 to 68, wherein each of the fixed bed supports comprises a protrusion or receiver for interlocking with a corresponding protrusion or receiver on an adjacent fixed bed support of the fixed bed support.

70. An apparatus for culturing cells, comprising:

a bioreactor comprising a housing having walls forming an interior compartment;

a fixed bed for culturing cells;

an annular fixed bed support for supporting a fixed bed, the fixed bed support adapted to be removably positioned within the interior compartment of the housing and defining a peripheral chamber between the annular fixed bed support and the housing;

An impeller for circulating a fluid through the fixed bed within the peripheral chamber; and

an impeller support for extending at least partially through the impeller and centering the impeller in the housing;

wherein the impeller is attached to the impeller support by a snap fit connection.

71. A method of manufacturing a fixed bed bioreactor comprising:

one or more fixed bed supports are interlocked within a single piece housing.

72. The method of example 71, wherein the interlocking step comprises interconnecting a first fixed bed support to a second fixed bed support.

73. The method of any one of examples 71-72, further comprising the step of positioning the impeller within a portion of the first fixed bed support.

74. The method of any of examples 71-73, wherein the interlocking step comprises interlocking the second fixed bed support with a cover for covering the housing.

75. The method of any of examples 71-74, further comprising the step of forming a first seal between the first fixed bed support and the second fixed bed support.

76. The method of example 75, further comprising the step of forming a second seal between the second fixed bed support and the housing.

77. The method of any one of examples 71-76, further comprising the step of wrapping the fixed bed around each of the fixed bed supports.

78. A bioreactor for culturing cells, comprising:

a housing having walls defining an interior compartment;

a plurality of fixed beds for culturing cells;

a plurality of annular fixed bed supports, each of the plurality of fixed bed supports adapted to support a respective at least one of a plurality of fixed beds, wherein each of the plurality of fixed bed supports comprises:

an annular section; and

a support frame extending radially outwardly from the annular section, the support frame having an outer diameter corresponding in size to an inner diameter of the loss wall of the housing, the support frame adapted to support at least one of the plurality of fixed beds from below and to allow fluid to flow through the support frame;

wherein the plurality of fixed bed supports are adapted to interlock with one another to form a peripheral chamber between the plurality of annular fixed bed supports and the wall of the housing and a central chamber within the annular section;

a cover for connecting to the housing and for sealing the plurality of fixed beds and the plurality of fixed bed supports in the interior compartment;

a plurality of probes extending into the interior compartment adjacent to or into at least one of the fixed beds;

an upper frame covering the plurality of fixed bed supports and forming a plurality of pockets for allowing fluid to accumulate therein upon exiting an upper end of the plurality of fixed beds, wherein at least one of the plurality of probes is adapted to sense a characteristic of the fluid in a respective one of the plurality of pockets;

An impeller for circulating a fluid within the bioreactor; and

a vessel for housing an impeller, the vessel comprising:

a plurality of openings adapted to allow fluid to flow from within the container to the peripheral chamber; and

a plurality of positioners in the form of radially extending arms extending therefrom and adapted to position the container within the housing and to space the container from its walls;

wherein the upper frame is adapted to interlock with at least one of the plurality of annular fixed bed supports and is adapted to interlock with the cover to prevent relative rotation therebetween.

79. A bioreactor for culturing cells, comprising:

a housing having walls defining an interior compartment;

a removable fixed bed for culturing cells;

a removable fixed bed support adapted to support a fixed bed, wherein the fixed bed support is annular in shape and comprises a plurality of arms extending radially outwardly, the radially extending arms defining an outer diameter corresponding in size to an inner diameter of a wall of the housing for positioning and centering the fixed bed support in the housing, wherein the plurality of arms are adapted to support the fixed bed from below;

wherein the fixed bed support forms a peripheral chamber between an outer wall of the fixed bed support and the housing, and a central chamber within the fixed bed support;

Wherein the fixed bed is adapted to be positioned within the peripheral chamber; and is also provided with

Wherein the housing includes one or more receptacles in a wall of the housing for receiving at least one of the plurality of arms, the one or more receptacles adapted to support the fixed bed support within the interior compartment and prevent relative rotation of the fixed bed support within the housing;

a cover for connecting to the housing and for sealing the fixed bed and fixed bed support in the interior compartment;

at least one probe extending into the interior compartment at a location within the peripheral chamber above the fixed bed;

an impeller adapted to rotate on an impeller support, the impeller for circulating a fluid within the bioreactor, the impeller being located in a chamber formed between a lower portion of the fixed bed support and a floor of the housing, wherein the impeller is adapted to circulate the fluid outwardly from a central chamber of the fixed bed support to a peripheral chamber and upwardly through the fixed bed therein; and

a drain connected to the impeller support for draining fluid from the bioreactor.

As used herein, the following terms have the following meanings:

as used herein, "a," "an," and "the" refer to both singular and plural referents unless the context clearly dictates otherwise. For example, "a/an compartment" refers to one or more compartments.

As used herein, "about," "substantially" or "approximately" refers to measurable values, such as parameters, amounts, time durations, etc., meaning variations comprising +/-20% or less, preferably +/-10% or less, more preferably +/-5% or less, even more preferably +/-1% or less, still more preferably +/-0.1% or less of particular values, which variations are suitable for execution in the disclosed invention so far. However, it is to be understood that the value referred to by the modifier "about" is also specifically disclosed per se.

As used herein, "comprising" and "having" are synonymous with "including" and are inclusive or open-ended terms that specify the presence of the following items, such as components, and do not preclude the presence or addition of additional, non-enumerated components, features, elements, components, steps, etc., known in the art or disclosed therein.

While various embodiments have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Many changes, modifications and substitutions will now occur to those skilled in the art without departing from the scope of the disclosure. For example, while the bioreactor is shown in a vertical orientation, it may be used in any orientation. In any of the foregoing embodiments, any or all of the components of the bioreactor 100, 200, 300 may be provided as disposable or "single use" components. This allows for inexpensive manufacture and use without the need for cleaning and re-sterilization. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. The following claims are intended to define the scope of protection under applicable law and to cover methods and structures within the scope of these claims and their equivalents.

Claims

1. An apparatus for culturing cells, comprising:

a bioreactor comprising a housing having walls forming an interior compartment;

a fixed bed for culturing cells;

a fixed bed support for the fixed bed, the fixed bed support adapted to be removably positioned within the interior compartment of the housing; and

2. The apparatus of claim 1, wherein the fixed bed support comprises a central portion and a peripheral portion extending radially outward from the central portion and including the one or more locators, the peripheral portion having an outer diameter corresponding to an inner diameter of the housing.

3. The apparatus of claim 2, wherein the peripheral portion comprises one or more protrusions.

4. A device according to claim 3, wherein the peripheral portion comprises an outer ring connected to the central portion by the one or more protrusions.

5. The apparatus of claim 2, wherein the peripheral portion comprises an annular disc-shaped surface having a plurality of apertures therein.

6. The apparatus of claim 2, wherein the peripheral portion comprises a mesh.

7. The apparatus of claim 2, wherein the peripheral portion is adapted to support the fixed bed from below.

8. The apparatus of claim 2, wherein the peripheral portion is adapted to allow fluid to flow therethrough.

9. The apparatus of claim 2, wherein the central portion includes a separator that forms a plurality of spaces in an interior of the central portion for receiving the conduits within the housing.

10. The apparatus of claim 9, wherein the plurality of spaces have different sizes.

11. The apparatus of claim 9, wherein at least one of the conduits is adapted to convey a fluid flow to or from the interior of the central portion.

12. The apparatus of claim 11, wherein the separator is adapted to position at least one of the conduits, the conduits being adapted to convey a fluid flow away from an inner wall of the central portion to prevent contact between the conduits and falling film fluid flowing down the inner wall.

13. The apparatus of claim 2, wherein the central portion of the support forms a receptacle for receiving an agitator.

14. The apparatus of claim 13 wherein the agitator comprises an impeller adapted to rotate about an agitator support, the agitator support adapted to receive and retain the fixed bed support.

15. The apparatus of claim 14, wherein the agitator support comprises a tubular post connected to a flexible drain connected to a lid of the bioreactor.

16. The apparatus of claim 15, wherein the peripheral portion of the fixed bed support comprises one or more protrusions in the form of a plurality of radially extending arms.

17. The apparatus of claim 16, wherein the plurality of radially extending arms are connected to a rim having an outer diameter corresponding to an inner diameter of the housing.

18. The apparatus of claim 16 wherein the plurality of radially extending arms engage and support the fixed bed.

19. The apparatus of claim 18, wherein the housing comprises a receiver for receiving at least one of the one or more protrusions.

20. The apparatus of claim 19 wherein the central portion and the peripheral portion of the fixed bed support comprise a single unitary structure.

21. The apparatus of claim 2, wherein the fixed bed comprises one or more layers of woven or nonwoven material wrapped around the central portion of the fixed bed support.

22. The apparatus of claim 1, further comprising a seal for sealing between the inner wall of the housing and the fixed bed support.

23. The apparatus of claim 1, wherein the fixed bed comprises a plurality of fixed bed sections, the fixed bed support further comprising a plurality of interlocking support sections for supporting each of the plurality of fixed bed sections.

24. The apparatus of claim 23, wherein each interlocking support section is adapted to interlock with an adjacent support section.

25. The apparatus of claim 23, further comprising a first seal for sealing each adjacent interlocking support member portion together.

26. The apparatus of claim 25, further comprising a second seal for sealing each of the plurality of portions to an inner wall of the housing.

27. The apparatus of claim 1, further comprising an upper frame for positioning over the fixed bed.

28. The apparatus of claim 27, wherein the upper frame has a height sufficient to form at least one pocket for allowing fluid to accumulate therein upon exiting an upper end of the fixed bed, and a sensor for sensing a characteristic of the fluid in the at least one pocket.

29. The apparatus of claim 27, wherein the upper frame is adapted to engage a cover of the bioreactor.

30. The apparatus of claim 27, wherein the upper frame forms a plurality of pockets, the plurality of pockets having different volumes.

31. The apparatus of claim 30, wherein the upper frame is adapted to receive or align one or more samplers for sampling the fixed bed.

32. The apparatus of claim 1, further comprising a cover removably connected to the housing.

33. The apparatus of claim 32 wherein the cover is adapted to hold the fixed bed vertically in place within the housing.

34. The apparatus of claim 33, wherein the cover comprises an overhang portion for engaging the fixed bed or the fixed bed support.

35. The apparatus of claim 32, wherein the cover is adapted to threadably engage the housing.

36. The apparatus of claim 32, further comprising a gasket between the cover and the housing.

37. The apparatus of claim 1, further comprising a port in the wall of the housing that is above an upper end of the fixed bed when positioned therein.

38. The apparatus of any one of claims 1 to 37, wherein the housing comprises a one-piece rigid structure forming the interior compartment.

39. An apparatus for culturing cells, comprising:

40. The apparatus of claim 39, wherein the assembly comprises an upper portion adapted to interlock with the cover.

41. The apparatus of claim 40, wherein the upper portion comprises an upper frame for positioning over the fixed bed.

42. The apparatus of claim 41 wherein the upper frame has a height sufficient to form at least one pocket for allowing fluid to accumulate therein upon exiting the upper end of the fixed bed, and a sensor for sensing a characteristic of the fluid in the at least one pocket.

43. The apparatus of claim 41, wherein the upper frame includes a recess for engaging a protrusion extending from a cover of the apparatus.

44. The apparatus of claim 41, wherein the upper frame forms a plurality of pockets, each having a different volume.

45. The apparatus of claim 41, wherein the fixed bed comprises a plurality of fixed bed sections, each fixed bed section being associated with one of a plurality of supports adapted to interlock with an adjacent support.

46. The apparatus of claim 45, wherein the upper frame is adapted to interlock with at least one of the plurality of supports.

47. The apparatus of claim 45, wherein each of the plurality of supports comprises a central portion and a peripheral portion, the peripheral portion adapted to allow fluid to reach the fixed bed, the peripheral portion having an outer diameter corresponding to an inner diameter of the housing.

48. The apparatus of claim 45, further comprising at least one O-ring located between at least two of the plurality of supports or between the cover and at least one of the plurality of supports.

49. The apparatus of claim 48, wherein the cover is adapted to provide downward pressure on the assembly when the cover is attached to the housing.

50. The apparatus of claim 49, wherein the downward pressure is sufficient to maintain the at least one O-ring in place without glue.

51. The apparatus of claim 39 wherein the assembly comprises a lower portion for receiving an agitator, the lower portion adapted to interlock with at least one support for supporting the fixed bed.

52. An apparatus for culturing cells, comprising:

a bioreactor comprising a housing having walls forming an interior compartment;

a fixed bed for culturing cells; and

a support for the fixed bed, the support adapted to be removably positioned within the interior compartment, the support at least partially forming a chamber housing an agitator and including one or more centering projections extending toward the wall of the housing.

53. The apparatus of claim 52, wherein the protrusions engage the fixed bed.

54. The apparatus of claim 52, wherein the housing includes one or more receptacles for receiving one or more of the protrusions.

55. The apparatus of claim 54, wherein engagement between one or more of the protrusions and one or more of the receptacles is adapted to prevent rotation of the support within the housing.

56. An apparatus for culturing cells, comprising:

a housing having walls forming an interior compartment;

a fixed bed for culturing cells within the interior compartment;

57. The apparatus of claim 56, wherein the upper frame includes one or more indicia for indicating a position or orientation of the one or more probes.

58. The apparatus of claim 56, wherein said one or more probes are attached to said upper frame.

59. An apparatus for culturing cells, comprising:

a one-piece housing;

at least one fixed bed for culturing cells; and

a plurality of fixed bed supports adapted to interlock for positioning within the single piece housing.

60. The apparatus of claim 59 wherein each of the plurality of fixed bed supports comprises an annular portion and a support frame extending radially outward from the annular portion.

61. The apparatus of claim 60, wherein the support frame has an outer diameter corresponding to an inner diameter of the housing.

62. The apparatus of claim 60, wherein the support frame comprises a generally planar extension.

63. The apparatus of claim 60, wherein the support frame is adapted to allow fluid to flow therethrough.

64. The apparatus of claim 60, wherein the support frame extends from a bottom of the annular portion.

65. The apparatus of claim 60, wherein the support frame comprises a plurality of radially extending arms connected to a peripheral ring.

66. The apparatus of claim 60, wherein the support frame comprises a mesh.

67. The apparatus of claim 60 wherein the support frame is adapted to support the at least one fixed bed from below.

68. The apparatus of claim 60 wherein the support frame is adapted to serve as a base for positioning the fixed bed on the fixed bed support.

69. The apparatus of claim 59, wherein each of the fixed bed supports comprises a protrusion or receiver for interlocking with a corresponding protrusion or receiver on an adjacent fixed bed support of the fixed bed support.

70. An apparatus for culturing cells, comprising:

a bioreactor comprising a housing having walls forming an interior compartment;

a fixed bed for culturing cells;

an annular fixed bed support for supporting the fixed bed, the fixed bed support adapted to be removably positioned within the interior compartment of the housing and defining a peripheral chamber between the annular fixed bed support and the housing;

71. A bioreactor for culturing cells, comprising:

a housing having walls defining an interior compartment;

a plurality of fixed beds for culturing cells;

a plurality of annular fixed bed supports, each of the plurality of fixed bed supports adapted to support a respective at least one of a plurality of the fixed beds, wherein each of the plurality of fixed bed supports comprises:

An annular section; and

a support frame extending radially outwardly from the annular section, the support frame having an outer diameter corresponding in size to an inner diameter of the wall of the housing, the support frame adapted to support at least one of the plurality of fixed beds from below and allow fluid to flow through the support frame;

wherein a plurality of said fixed bed supports are adapted to interlock with one another to form a peripheral chamber between a plurality of annular said fixed bed supports and said wall of said housing and a central chamber within said annular section;

an upper frame covering a plurality of said fixed bed supports and forming a plurality of pockets for allowing fluid to accumulate therein upon exiting an upper end of a plurality of said fixed beds, wherein at least one of a plurality of said probes is adapted to sense a characteristic of the fluid in a respective one of a plurality of said pockets;

an impeller for circulating a fluid within the bioreactor; and

A container for housing the impeller, the container comprising:

a plurality of locators in the form of radially extending arms extending therefrom and adapted to locate the container within the housing and to space the container from the walls thereof;

wherein the upper frame is adapted to interlock with at least one of the plurality of annular fixed bed supports and to interlock with the cover to prevent relative rotation therebetween.

72. A bioreactor for culturing cells, comprising:

a housing having walls defining an interior compartment;

a removable fixed bed for culturing cells;

a removable fixed bed support adapted to support the fixed bed, wherein the fixed bed support is annular in shape and comprises a plurality of arms extending radially outwardly, the radially extending arms defining an outer diameter corresponding in size to an inner diameter of the wall of the housing for positioning and centering the fixed bed support in the housing, wherein the plurality of arms are adapted to support the fixed bed from below;

Wherein the housing comprises one or more receptacles in the wall of the housing for receiving at least one of a plurality of the arms, one or more of the receptacles adapted to support the fixed bed support within the interior compartment and prevent relative rotation of the fixed bed support within the housing;

a cover for connecting to the housing and for sealing the fixed bed and the fixed bed support in the interior compartment;

at least one probe extending into the interior compartment at a position within the peripheral chamber above the stationary bed;

an impeller adapted to rotate on an impeller support for circulating a fluid within the bioreactor, the impeller being located in a chamber formed between a lower portion of the fixed bed support and a floor of the housing, wherein the impeller is adapted to circulate fluid outwardly from the central chamber of the fixed bed support to the peripheral chamber and upwardly through the fixed bed therein; and

a drain connected to the impeller support for draining the fluid from the bioreactor.