Biological modeling device

Abstract

The present invention provides a biological modeling device for co-culturing cells, comprising: a first chamber providing a first reservoir; a second chamber providing a second reservoir, where the second reservoir and the first reservoir are connected by a permeable support, and the first chamber is at least partially accommodated into the second chamber; and a seal arranged and sealingly engaging the first and second chambers such that a first gaseous environment of the first reservoir of the first chamber is separated from a second gaseous environment of the second reservoir of the second chamber by the seal.

Description

Technical Field

[0001] The present invention generally relates to a biological modeling device. In particular, the present invention relates to a biological modeling device that includes two chambers (each having a gaseous environment), with an improved sealing arrangement for maintaining isolation between the gaseous environments of the two chambers. Background Art

[0002] Known biological modeling devices are used for in vitro culturing of biological cells and / or tissues in order to simulate in vivo environments, such as, for example, a luminal model of the digestive tract. Typically, a biological modeling device has a plate with one or more notches or "wells" that are used to accommodate biological elements (such as cell populations or tissue samples) on the apical side. Some devices also have one or more cup-shaped inserts, each placed within a well. The inserts accommodate and hold different biological elements (such as a second cell population or tissue sample) on the basal side. Some known inserts have a membrane on the bottom surface. When a nutrient medium is introduced into one or more wells of the plate and / or the inserts, the biological element(s) obtain nutrients from the culture medium.

[0003] There is a desire to provide a biological modeling device with an improved sealing arrangement that is capable of maintaining isolation between the gaseous environments (plural gaseous environments) of two chambers. Specifically, an object of the present invention is to provide a biological modeling device that provides a sealed and separated gaseous environment between a well and an insert. An object of the present invention is to provide a biological modeling device in which the gaseous transmission path between the well(s) and the insert(s) substantially passes through an adjacent membrane.

[0004] The present invention provides at least an alternative to the prior art biological modeling devices. Summary of the Invention

[0005] According to the present invention, there is provided a biological modeling device according to the appended claims.

[0006] According to one aspect of the present disclosure, there is provided a biological modeling device that includes two chambers: a first chamber that provides a first reservoir having a first gaseous environment; a second chamber that provides a second reservoir having a second gaseous environment, wherein the second reservoir and the first reservoir are connected by a permeable support, and the first chamber is at least partially received within the second chamber; and a seal that is arranged and sealingly engages the first and second chambers such that the first gaseous environment of the first reservoir and the second gaseous environment of the second reservoir are separated by the seal.

[0007] In certain embodiments, the permeable support is a membrane.

[0008] In some embodiments, the permeable support is at least a gas-permeable membrane.

[0009] In some embodiments, the permeable support is a porous surface that allows gas to be transferred between the first and second reservoirs.

[0010] In some embodiments, the permeable support is a porous surface that allows at least one or more of gas, nutrients, and metabolites to pass between the first and second reservoirs.

[0011] In some embodiments, the seal prevents mixing of the first gaseous environment and the second gaseous environment except through the permeable support. In this way, the device provides two separated gaseous environments in the first and second chambers, which are connected only by the permeable support for the purpose of gaseous and / or nutrient transfer between the two chambers. In this way, the biomodeling device is provided with a sealed arrangement for maintaining sealed and different (i.e., separately controllable) gaseous environments in the first and second reservoirs. This configuration enables introduction into, and sampling from, multiple chambers with different gaseous environments without disturbing or mixing the gaseous environments. Gas can be transferred through the permeable support. The gas diffuses / transfers through biological elements (e.g., through biomaterials / cells) supported by the permeable support. By providing two gaseous environments that are sealed from each other except through the permeable support, the device is operable for biologically modeling anaerobic and aerobic separation conditions, and / or testing the passage of smoke, airborne toxins, drugs, etc. through biological elements (such as a population of colon cells, a population of epithelial cells, an in vitro airway model, etc. held on the permeable support).

[0012] As used herein, a "primary path" or "predominant path" refers to the path through which substantially all gaseous transport occurs. In some embodiments, gas flow between the first reservoir and the second reservoir occurs through the permeable support as the primary path. Any inherent permeability of the sealing material is small compared to the gas permeability of the permeable support.

[0013] In some embodiments, the first gaseous environment includes the headspace of the first reservoir of the first chamber.

[0014] In some embodiments, the second gaseous environment includes the headspace of the second reservoir of the second chamber.

[0015] As used herein, the "headspace" of a reservoir refers to the gaseous space above the biological elements and / or any solution in the reservoir.

[0016] In certain embodiments, a bioremodeling device can be used for co-culturing cells. In such a system, the first and second chambers each include a biological element in the form of a cell (or cells) population, and the primary path between the chambers is through a permeable support. Specifically, a flow path is provided through the permeable support as the primary (i.e., first) path. A particular advantage of this arrangement is that, for example, an aerobic environment can be used to propagate one cell population while an anaerobic environment can be used simultaneously to propagate another cell population. Thus, each environment can be controlled separately, independent of the other.

[0017] In certain embodiments, the second gaseous environment is sealed from the first gaseous environment by a combination of a seal and the first chamber.

[0018] In certain embodiments, the first gaseous environment is an anaerobic environment.

[0019] In certain embodiments, the second gaseous environment is an aerobic environment.

[0020] In certain embodiments, the first and / or second reservoir is configured to contain and retain at least one biological element. In certain embodiments, the biological element can be one or more of a mixed cell population, a single cell population, a cell co-culture, a bacterial culture, a microbiota, mucus, etc.

[0021] Additionally or alternatively, the first and / or second reservoir is configured to contain and retain a solution. In certain embodiments, the solution can be one or more of a culture or nutrient medium, a buffer solution, a test solution, etc.

[0022] In certain embodiments, the seal is a gasket seal.

[0023] In certain embodiments, the permeable support is integrally formed with the gasket seal. In such embodiments, a separate first chamber element is not required and the gasket seal can form the first chamber.

[0024] In certain embodiments, the gasket seal includes a penetrable portion.

[0025] In certain embodiments, the penetrable portion includes a septum.

[0026] In certain embodiments, the septum is a resealable or self-sealing septum. In this way, a sampling or injection device can be introduced into the first or second chamber independent of the other chamber. The gaseous environment of the chamber receiving the sampling or injection device remains sealed off from the other gaseous environment.

[0027] In certain embodiments, the first chamber is an insert including a permeable support.

[0028] In certain embodiments, the permeable support forms at least a part of the bottom surface of the first chamber.

[0029] In some embodiments, the insert includes a wall that stands upright from the bottom surface of the permeable support.

[0030] In some embodiments, the first chamber is partially received into the second chamber through a gasket seal.

[0031] In some embodiments, the seal includes a barrier layer that is arranged to sealingly engage with the second chamber so as to separate the second gaseous environment from the first gaseous environment.

[0032] In some embodiments, the barrier layer includes an opening.

[0033] In some embodiments, the first chamber is partially received through the opening to form a sealing engagement between the barrier layer and the first chamber, such that the second chamber is sealed by the combination of the barrier layer and the first chamber.

[0034] In some embodiments, the opening of the barrier layer is a self-sealing diaphragm. In this way, when the first chamber is pressed and passed through the opening, the barrier layer forms a sealing engagement with the first chamber for the second chamber.

[0035] In some embodiments, the first chamber is part of the barrier layer.

[0036] In some embodiments, the first chamber is integrally formed with the barrier layer.

[0037] In some embodiments, the permeable support is integrally formed with the barrier layer. In such embodiments, a separate first chamber element is not required, and the barrier layer can form the first chamber.

[0038] In some embodiments, the second chamber is located in the substrate. In some embodiments, the second chamber is integrally formed with the substrate.

[0039] In some embodiments, the second chamber includes a bottom surface, an inner wall extending upward from the bottom surface toward an open top, and the bottom surface and the inner wall together define a second reservoir.

[0040] In some embodiments, the biomodeling device includes a plurality of second chambers, and each second chamber includes a second reservoir.

[0041] In some embodiments, the biomodeling device includes a plurality of first chambers, each first chamber corresponding to each of the plurality of second chambers, each first chamber including a first reservoir, and a permeable support located between the respective first and second reservoirs.

[0042] In some embodiments, each first chamber is an insert.

[0043] In some embodiments, each first chamber is integrally formed with a seal, and wherein each first chamber is integrally formed with a permeable support.

[0044] In some embodiments, the biologic modeling device includes a plurality of seals, each seal disposed between and sealingly engaging a respective first chamber and a second chamber.

[0045] In some embodiments, the seal is disposed between an outer surface of the first chamber and an inner wall of the second chamber and sealingly engages the two.

[0046] In some embodiments, the biologic modeling device further includes a lid for the first chamber (or each first chamber), wherein the first chamber is enclosed by the lid.

[0047] In some embodiments, the lid includes a seal that is arranged to sealingly engage the first chamber so as to separate the first gaseous environment from the surrounding ambient environment, and wherein the first chamber is sealed by the seal.

[0048] In some embodiments, the seal of the lid includes a penetrable portion.

[0049] In some embodiments, the penetrable portion includes a diaphragm.

[0050] In some embodiments, the diaphragm is a resealable or self-sealing diaphragm.

[0051] In some embodiments, the seal of the lid includes more than one (preferably two) penetrable portions.

[0052] In some embodiments, one penetrable portion is configured to allow access to one of the first chamber or the second chamber, while the other penetrable portion is configured to allow access to the other of the first chamber and the second chamber.

[0053] In some embodiments, each penetrable portion includes a diaphragm.

[0054] In some embodiments, the diaphragm is a resealable or self-sealing diaphragm.

[0055] In some embodiments, the biologic modeling device further includes a transfer lid. More specifically, the transfer lid is formed of a gas-permeable material. Even more specifically, the transfer lid is configured and arranged to cover at least the first chamber or each of a plurality of first chambers. In this way, the biological elements (multiple biological elements) and / or solutions (multiple solutions) in the first chamber are prevented from transferring between adjacent first chambers.

[0056] In some embodiments, the transfer lid is a gas-permeable silicon membrane.

[0057] In some embodiments, the first chamber is a top chamber.

[0058] In some embodiments, the second chamber is a basolateral chamber.

[0059] In some embodiments, the first reservoir is configured to contain and hold a first culture medium for feeding a first biological element. In some embodiments, the second reservoir is configured to contain and hold a second culture medium for feeding the first and / or second biological elements.

[0060] In some embodiments, the second reservoir is configured to contain and hold a second culture medium for maintaining a first biological element through a permeable support. More specifically, the second culture medium can provide nutrients for the first biological element through the permeable support.

[0061] In some embodiments, the first chamber includes a seal plug that operably seals the first reservoir from the surrounding environment. In this way, the first chamber can be closed and not used.

[0062] In some embodiments, the biomodeling device is a culture device for growing cells and / or tissues in vitro.

[0063] According to another aspect of the present disclosure, a method for growing cells and / or tissues in vitro using a biomodeling device according to another aspect of the present invention is provided.

[0064] According to still another aspect of the present disclosure, a biomodeling device is provided, comprising: a first chamber providing a first reservoir with a first gaseous environment; a second chamber having a second gaseous environment, wherein the second reservoir and the first reservoir are connected by a permeable support, and the first chamber is at least partially received in the second chamber; and a seal arranged and sealingly joining the first and second chambers such that the first gaseous environment of the first reservoir of the first chamber and the second gaseous environment of the second reservoir of the second chamber are separated by the seal; and a lid for the first chamber, wherein the first chamber is closed by the lid, and wherein the lid includes at least one opening and a conduit extending from the at least one opening towards the first chamber.

[0065] In some embodiments, the lid further includes at least one opening and a conduit extending from the at least one opening towards the second chamber.

[0066] In some embodiments, the first reservoir is configured to contain and hold a first biological element.

[0067] In some embodiments, the second reservoir is configured to contain and hold a second biological element.

[0068] In some embodiments, the first and / or second biological element(s) may include a cell population. More specifically, the cell population may be selected from one or more of the following: bacteria, epithelial cells, immune cells.

[0069] In some embodiments, the first and / or second reservoir(s) contain(s) and hold(s) a solution. More specifically, the solution may be one or more of a culture or nutrient medium, a buffer solution, a test solution, etc.

[0070] According to another aspect of the present disclosure, there is provided a biological modeling device, comprising: a first chamber providing a first reservoir having a first gaseous environment; a second chamber providing a second reservoir having a second gaseous environment, wherein the second reservoir and the first reservoir are connected by a permeable support, and the first chamber is at least partially received in the second chamber; and a seal disposed and sealingly joining the first and second chambers such that the first gaseous environment of the first reservoir of the first chamber and the second gaseous environment of the second reservoir of the second chamber are separated by the seal; and a lid for the first chamber, wherein the first chamber is enclosed by the lid, and wherein the lid includes at least one opening and a conduit extending from the at least one opening toward the second chamber.

[0071] In some embodiments, the lid further includes at least one opening and a conduit extending from the at least one opening toward the first chamber.

[0072] In some embodiments, the first reservoir is configured to contain and hold a first biological element.

[0073] In some embodiments, the second reservoir is configured to contain and hold a second biological element.

[0074] In some embodiments, the first and / or second biological element(s) may include a cell population. More specifically, the cell population may be selected from one or more of the following: bacteria, epithelial cells, immune cells.

[0075] In some embodiments, the first and / or second reservoir(s) contain(s) and hold(s) a solution. More specifically, the solution may be one or more of a culture or nutrient medium, a buffer solution, a test solution, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0076] Embodiments of the present invention will now be described by way of example only with reference to the accompanying drawings, in which: Figure 1 An example of a biological modeling device is shown, in which: (a) is an exploded perspective view; (b) is an exploded perspective view seen from the front; (c) is a perspective view of the assembled configuration; (d) is a cross-sectional view seen from the front; and (e) is an enlarged cross-sectional view as seen from the front; Figure 2 Another example of a biomodeling device is shown, wherein: (a) is an exploded perspective view; (b) is an exploded perspective view as seen from the front; (c) is a perspective view of the assembled configuration; (d) is a cross-sectional view as seen from the front; and (e) is an enlarged cross-sectional view as seen from the front; Figure 3 shows Figure 2 the biomodeling device in (a) is an exploded perspective view; (b) is an exploded perspective view as seen from the front; (c) is a perspective view of the assembled configuration; (d) is a cross-sectional view as seen from the front; and (e) is an enlarged cross-sectional view as seen from the front; Figure 4 Another example of a biomodeling device is shown, wherein: (a) is an exploded perspective view as seen from the top; (b) is an exploded perspective view as seen from the bottom; and (c) is a cross-sectional view of the assembled configuration; Figure 5 Another example of a biomodeling device is shown, which is an enlarged view of an insert, wherein: (a) is an exploded perspective view; (b) is a partially exploded perspective view; (c) is a perspective view of the assembled configuration; and (d) is a cross-sectional view of the assembled configuration; Figure 6 A part of a substrate is shown, wherein: (a) is a perspective view; and (b) is a cross-sectional view; Figure 7 The substrate of a biomodeling device printed using (a) fused deposition modeling (FDM); and (b) stereolithography (SLA) is shown; Figure 8 A perspective view of (a) the substrate of a biomodeling device; (b) a perspective view of an insert tray of a biomodeling device; (c) a perspective view of a substrate and insert tray assembly; (d) a perspective view of a substrate, insert tray, and lid assembly; and (e) a cross-sectional view of an assembly including a substrate and an insert tray is shown; Figure 9 A biomodeling device model fabricated using stereolithography 3D printing is shown: (a) the substrate, insert tray, and lid are separated; (b) the substrate and insert tray are assembled together and the lid is separated; and (c) an assembly of the substrate, insert tray, and lid; Figure 10 A biomodeling device is shown, including a transfer lid configured and arranged to prevent material transfer between adjacent first chambers. Detailed Description

[0077] Certain terms used in the following description are for convenience only and are not limiting. Words such as "right", "left", "lower", "upper", "front", "rear", "upward", "down", and "downward" denote directions with reference to the accompanying drawings and are relative to the components described when assembled and installed. "Inner", "inward" and "outer", "outward" each refer to the direction towards and away from the designated center line or geometric center (such as the central axis) of the described element, and the specific meaning can be easily seen from the context of the description.

[0078] In addition, as used herein, the terms "connect", "attach", "couple", "mount" are intended to include both direct connections between two members without any other members inserted therebetween, and indirect connections with one or more other members inserted therebetween. The terms include the specifically mentioned words above, their derivatives, and words with similar meanings.

[0079] In addition, unless otherwise specified, the use of ordinal adjectives such as "first", "second", "third", etc. only indicates different instances of the similar objects mentioned, and is not intended to imply that the objects so described must have a given order in time, space, sequence, or in any other way.

[0080] Like reference numerals are used to denote like features throughout the text.

[0081] Now referring to Figure 1 , a biomodeling device 2 is shown. The biomodeling device 2 is provided with a number of inserts (i.e., first chambers) 4 which, in this example, are inserted through the openings in the tray 27. In the illustrated embodiment, the inserts 4 are provided with outwardly extending arms 5 which engage the surface of the tray 27 to hold the inserts 4 in place. Each insert 4 has a tubular inner wall 8 which terminates at one end in a bottom surface 6 and is open at the other end. The tray 27 forms an airtight seal around each insert 4. A porous membrane 20 is integrally formed with the bottom surface 6 of each insert 4. The insert 4 defines a reservoir for holding the biological material to be cultured (such as a cell population), such as a population containing bacterial cells. In this example, the insert 4 is a tip chamber. The biomodeling device 2 is adapted for culturing cells and / or tissues in vitro. The pore size of the membrane 20 is between 0.4 and 8 microns. In a specific example, to model the intestine, a membrane 20 with a pore size of 1.8 microns is used. The membrane 20 has a scaffold structure that can promote the tissue-like behavior of the cultured cells. For example, the scaffold structure can be a gel or other tissue culture matrix.

[0082] The device 2 is provided with slots (i.e., second chambers) 12, each of which is formed as a portion of the substrate 24. The number of slots 12 corresponds to the number of inserts 4 in the tray 27. In this example, twenty-four slots 12 are provided, arranged in a six-by-four matrix, but any number of slots 12 can be considered depending on the requirements of cell propagation, such as, for example, six, twelve, twenty-four, forty-eight, ninety-six, three hundred and eighty-four, or one thousand five hundred and thirty-six slots 12. Each slot 12 has a tubular inner wall 16, one end of which terminates at the bottom surface 14 and the other end is an open top. The inserts 4 are arranged in the tray 27 to be aligned with the corresponding slots 12 of the substrate 24 such that the central axis of each insert 4 is aligned with the central axis of each slot 12. The diameter of the insert 4 is smaller than the diameter of the corresponding tubular slot 12. Each slot 12 defines a reservoir for holding biological material (such as a group containing epithelial cells). In this example, the inserts 4 and the slots 12 are tubular, but other regular and irregular shapes can also be considered. In this example, the slots 12 are basal side chambers.

[0083] A seal 22 is provided around the insert or each insert 4. In this example, separate seals 22 are provided around each insert 4. The seal 22 is sized such that the insert 4 of the tray 27 is received through the seal 22 and placed into the slot 12 of the substrate 24. When the insert 4 is received into the respective slot 12 in this way, the seal 22 engages with the insert 4 and the slot 12, thereby providing a sealed arrangement that separates the gaseous environment within the top space of the insert 4 from the gaseous environment within the top space of the slot 12. Specifically, the seal 22 is arranged between the outer surface of the insert 4 and the inner wall 16 of the slot 12. The top space 18 of the slot 12 is enclosed by the seal 22 and the insert 4, thereby providing a seal. In this specific example, a gasket 23 is provided to improve the seal between the insert 4 and the slot 12, which is most clearly seen in Figure 1 (e).

[0084] The substrate 24 is provided with a skirt 26 that extends continuously around the perimeter of the substrate 24. The tray 27 is provided with a flange 28 that extends outwardly from the tray 27. The tray 27 is sized and shaped such that the flange 28 of the tray 27 is retained within the substrate 24 by the skirt 26 of the substrate 24. When the tray 27 is held within the substrate 24 in this way, each insert 4 is aligned with and inserted into its respective slot 12. At the same time, the seal 22 contacts the inner wall 16 of the slot 12 to provide a sealed environment within the reservoir of the slot 12. The seal 22 is formed together with a barrier layer 30 that cooperates with the insert 4 to seal the slot 12. One surface of the substrate 24 is provided with an air valve 25 that allows air to freely flow from the surrounding environment surrounding the outside of the device 2 into the substrate 24, and more specifically, into the slot 12. This maintains an aerobic environment in the slot 12. It is contemplated that the air valve 25 can be provided with a particulate filter.

[0085] The reservoir of the insert or each insert 4 is connected to the respective trough 12 by a porous membrane 20. In this example, the porous membrane 20 is integrally formed with the bottom surface 6, but it is also conceivable to place the porous membrane 20 as a separate component on the bottom surface 6 of the insert 4. The porous membrane 20 can be provided in the form of a mesh or a matrix of pores. The porosity of the membrane 20 allows, for example, gases, nutrients, and metabolites to pass between the respective reservoirs of the insert 4 and the trough 12.

[0086] The bioreactor device 2 is provided with a lid 32 which, when in the closed position, encloses the top space 10 of the insert 4. The lower part of the lid 32 is provided with a flange 34 which extends around the periphery of the lid 32. A seal 38 is arranged to engage with the flange such that when the lid 32 is closed, the lid 32 seals against the tray 27 and the substrate 24. The seal 38 can be a gasket seal. Each of the opposite outer surfaces of the lid 32 has a latch 36 which is attached to the surface of the lid 32 by a hinge such that each latch 36 can pivot independently about its hinge relative to the lid 32.

[0087] In use, referring to Figure 1 (c) to (e), each insert 4 is provided with a first biological element (such as bacterial cells) and a maintenance medium, such as phosphate buffered saline (PBS). Each trough 12 is provided with a second biological element (such as epithelial cells) and a different culture medium for culturing the cells. The tray 27 is lowered into the substrate 24 such that each insert 4 is lowered into and received by its respective trough 12, and the porous membrane 20 is inserted between the reservoirs of the insert 4 and the trough 12. When the tray 27 is fully received within the substrate 24, the flange 28 of the tray 27 is retained by the skirt 26 of the substrate 24. In this arrangement, the seal 22 engages with the inner wall 16 of the trough 12 and with the insert 4 such that the top space 18 of the trough 12 is enclosed by the seal 22 and the insert 4. Thus, the gaseous environment within the top space 10 of the insert 4 and the gaseous environment within the top space 18 of the trough 12 are separated by the seal 22. At the same time, the porous membrane 20 provides a path for the transfer of gases, nutrients, and / or metabolites between the respective reservoirs of the insert 4 and the trough 12.

[0088] When the tray 27 and the substrate 24 are assembled together to receive the insert 4 within the trough 12, the top space 10 of each insert 4 is open to the surrounding environment. To enclose the top space 10 of the insert 4 from the environment, the lid 32 is placed on the tray 27 and the latches 36 are pivoted about their hinges to engage with the bottom surface of the skirt 26.

[0089] The lower chamber (i.e., trough 12) may be referred to as the basolateral chamber, and the upper chamber (i.e., insert 4) may be referred to as the apical chamber. In use, the upper chamber may include bacterial cells cultured in an anaerobic environment. A sachet may be provided within the chamber to create an anaerobic environment. The contents of the sachet initiate a reaction that quenches the oxygen in the environment. The lower chamber may include mammalian epithelial cells cultured in an aerobic environment. In this example, the aerobic environment may be provided by an air valve 25 that allows air to freely flow into the lower chamber. However, in other examples, it may be considered that both the upper and lower chambers are supplied with gas from an external gas supply. Thus, the gaseous environment of the upper chamber is separated from the gaseous environment of the lower chamber to provide a controlled way to co-culture cells in separate gaseous environments. The upper chamber may be used to simulate the lumen. The lower chamber may be used to simulate the intestinal wall. It may be considered to additionally form a mucus layer on the porous membrane 20. Specifically, it may be considered to form a mucus layer on top of the epithelial cells to simulate the intestinal epithelium. It may also be considered that if some inserts 4 are not used, the open top of the insert 4 may be sealed with a sealing plug (not shown). In this way, the excess inserts 4 are enclosed and not used.

[0090] Figure 2 Another example of the biomodeling device 2 is shown, where the trough 12, the seal 22, and the lid 32 are different from those in the Figure 1 example shown. The biomodeling device 2 is provided with a plurality of troughs 12 formed as part of the substrate 24. In this embodiment, each trough 12 has an inner wall 16 with a square or rectangular footprint that terminates at the bottom surface 14, and an open top opposite the bottom surface 14. Each trough 12 defines a reservoir for holding biological materials such as epithelial cells. The biomodeling device 2 is provided with inserts 4 in the same way as Figure 1 before and will not be described in detail here.

[0091] A seal 22 is provided around the insert or each insert 4 such that the insert 4 is received through the seal 22 and placed into the trough 12 of the substrate 24. In this example, the seal 22 has a frustoconical outer profile that tapers inwardly towards the substrate 24. In use, when the tray 27 is received onto the substrate 24, the inserts 4 descend into their respective troughs 12. When the inserts 4 descend into the troughs 12 in this way, the lower end of the seal 22 engages with the inner wall 16 of the trough 12 such that the top space 18 of the trough 12 is enclosed and sealed by the seal 22 and the respective inserts 4. It may be considered that the seal 22 may have a penetrable portion. The penetrable portion may have a resealable diaphragm. In a specific example, it may be considered that the penetrable portion of the seal 22 may be a self-sealing diaphragm.

[0092] The biomodeling device 2 is provided with a lid 32 which, when in the closed position, encloses the top space 10 of the insert 4. In this example, the lid 32 is different from the embodiment in Figure 1 in that it is provided with an array of first openings 40 and second openings 42. The diameter of the first openings 40 is greater than the diameter of the second openings 42. The first openings 40 are arranged in a six-by-four matrix which is aligned with the central axis of the respective insert 4. Each of the second openings 42 is arranged offset from the first openings 40 and is thus also offset from the central axis of the respective insert 4. Each of the first openings 40 is covered by a penetrable film 41. Similarly, each of the second openings 42 is covered by a penetrable film 43. The penetrable films 41, 43 have resealable diaphragms. More specifically, the penetrable films 41, 43 have self-sealing diaphragms.

[0093] As shown in Figure 2 (d) and 2(e), each of the first openings 40 leads to a conduit 44 which provides a passage leading to the respective aligned insert 4 for introducing / extracting culture medium and / or cells from the insert 4 when the film 41 is penetrated. In this example, each of the second openings 42 leads to a separate conduit 46 which provides a passage leading to the respective trough 12 for introducing / extracting culture medium and / or cells from the trough 12 when the film 43 is penetrated. Although in this example the conduits 44, 46 are shown extending linearly downwards, in other examples it may be considered that the conduits 44, 46 may instead extend towards the insert 4 and / or the trough 12 in different directions or in a non-linear manner. For example, the conduits 44, 46 may extend at an angle of 20 degrees offset from the central axis of the trough 12. In an arrangement not shown, the conduits 44, 46 extend into the insert 4 and serve as condensate collectors. In this way, condensate can be collected and the transmission of biological material (such as bacteria) between adjacent inserts 4 can be prevented. Thus, the sterility of the device can be improved. The gas permeability of the conduits 44, 46 ensures a continuous first gaseous environment between adjacent upper chambers (such as the insert 4).

[0094] In this example, each of the trough 12, the seal 22 and the lid 32 is modified from the example in Figure 1 but other examples are also considered where one or more (but not all) of the trough 12, the seal 22 and the lid 32 are modified from the example in Figure 1 For example, it is considered that the biomodeling device 2 may include all the features of Figure 1 where the lid 32 is changed to include the openings 40, 42 and the respective penetrable films 41, 43. Different examples of the biomodeling device 2 may include a lid 32 having the openings 40, 42, the penetrable films 41, 43 and the conduits 44, 46.

[0095] Figure 3shows another example of the bioreactor device 2, which is substantially the same as the device 2 in Figure 2 . However, the larger first opening 40 is not sealed by the penetrable film 41. Similarly, the smaller second opening 42 is not sealed by the penetrable film 43. However, the openings 40, 42 are through holes extending through the entire thickness of the lid 32. Figure 3 The bioreactor device 2 in Figure 3 is provided with a sheet 50 having openings 52, 54 arranged to align with the larger first opening 40 and the smaller second opening 42 in the lid 32, respectively. Specifically, the size and spacing of the opening 52 of the sheet 50 correspond to and align with the larger first opening 40 of the lid 32. The size and spacing of the opening 54 of the sheet 50 correspond to and align with the smaller second opening 42 of the lid 32. A penetrable base layer 48 is disposed between the lid 32 and the sheet 50. The size and arrangement of the penetrable base layer 48 are such that it covers and extends between the openings 40, 42 of the lid 32 and the openings 52, 54 of the sheet 50.

[0096] In this example, the penetrable base layer 48 has a resealable diaphragm. It is contemplated that the penetrable base layer 48 may have a self-sealing diaphragm. In use, the base layer 48 is penetrated through the opening 52 to access the underlying insert 4 through the conduit 44 to introduce / extract culture medium and / or cells from the insert 4. The base layer 48 is penetrated through the opening 54 to access the underlying trough 12 through the conduit 46 to introduce / extract culture medium and / or cells from the trough 12.

[0097] Figure 4 shows an embodiment of the bioreactor device 2 having an alternative sealing arrangement to the previous example. Each insert 4 of the tray 27 is provided with a seal in the form of an O-ring 22 that is received around the insert 4 and abuts the lower surface of the tray 27. When the insert 4 is received in the corresponding trough 12 of the substrate 24, the chamber of the trough 12 is separated from the chamber of the insert 4. The O-ring 22 provides a liquid-tight and gas-tight seal between the insert 4 and the trough 12. The substrate 25 is provided with an air valve 25 that can be connected to an external tube or pipe for introducing gas or fluid into the trough 12. Since the O-ring 22 is provided above the chamber of the trough 12, gas can pass through the entire substrate 24 and into the interior of the trough 12 during use. In this example, the O-ring 22 is made of nitrile rubber and has an inner diameter of 14 mm and an outer diameter of 18 mm.

[0098] Figure 5Shows a further modification to the bioreactor device 2, where the porous membrane 20 is attached to the bottom of the insert 4 and held in place by a clamp 58. The insert 4 is provided with protrusions 56 on opposite sides of the lower surface. The protrusions 56 engage with corresponding channels (i.e., cutouts) 60 in the clamp that hold the clamp in place around the outer wall of the insert 4. The porous membrane 20 is sized larger than the opening of the clamp 58 such that the porous membrane 20 abuts against the insert 4 and retains it in position. By providing the clamp 58 in this way, there is no need to use an adhesive to attach the porous membrane 20 in place. In this example, the gap between the bottom of the clamp 58 and the bottom of the groove 12 is 2 millimeters (mm).

[0099] Figure 6 Shows an alternative embodiment, which shows a portion of the substrate 24 with a single groove 12. The groove 12 is provided with a recess (i.e., slot) 62 at the distal end that houses an O-ring seal. By providing an O-ring within the recess 62, a seal is formed between the substrate 24 and the insert when the insert is received within the groove 12.

[0100] Figure 7 Shows two different samples of the substrate 24 produced using different 3D printing techniques. Figure 7 (a) Shows the substrate 24 printed using fused deposition modeling (FDM) 3D printing with acrylonitrile butadiene styrene (ABS) as the printing material. Figure 7 (b) Shows the substrate 24 printed using stereolithography (SLA) 3D printing with Biomed clear resin as the printing material.

[0101] Figure 8 Shows various components of the bioreactor device. Figure 8 (a) Shows the substrate 24 with eight tubular grooves 12 and valves 25 at either end that are used to introduce gas or fluid into the grooves 12 during cell culture. Figure 8 (b) Shows a tray 27 that includes eight inserts 8, the inserts being shaped and sized to be retained within the grooves 12. In use, the substrate 24 and the tray 27 are assembled together, and optionally, as Figure 8 (d) Shows, a lid 32 is provided to prevent evaporation of the contents within the device.

[0102] As Figure 9As shown, each of the substrate 24, the insert tray 27, and the lid 32 can be manufactured separately (e.g., by 3D printing). In this specific example, the components are 3D printed using stereolithography with acrylonitrile-butadiene-styrene (ABS) as the printing material. To assemble the bioprinting device 2, the tray 27 is placed on the substrate 24 such that the insert 4 is held within the corresponding slot 12. Then the lid 32 is placed on top of the tray 27 and substrate 24 assembly to prevent evaporation of the internal contents during cell culture. The substrate 24 is provided with a valve 25 that can be attached to an external conduit or tube (not shown) for introducing fluid or gas into the chamber of the slot 12. For example, oxygen can be introduced through the valve 25 to create an aerobic environment within the slot 12.

[0103] Figure 10 The bioprinting device 2 with a gas-permeable silicon transfer lid 64 is shown. The transfer lid 64 is located on top of the insert tray 27 and prevents the transfer of biomaterials and / or solutions between adjacent inserts 4. The gas permeability of the transfer lid 64 ensures a continuous first gaseous environment between adjacent upper chambers (e.g., inserts 4).

[0104] The bioprinting device of the present invention is suitable for use in a variety of exemplary models. The following models and biomaterials are provided only as examples. Human colon model

[0105] Representative colon cell lines (such as CACO-2 or T84) are grown in a monolayer on a transwell insert 4. In the first chamber provided by the insert 4, a cell-compatible mucus layer is placed on top of the cell layer. A bacterial inoculum and a nutrient medium are placed above the mucus layer. The bacterial inoculum can be one of the following: a single bacterium, multiple bacteria, or a representative microbial culture grown from a fecal inoculum.

[0106] The upper chamber provided by the insert 4 has an anaerobic environment (oxygen content less than about 0.5-1%) that is generated by GasPak TM The lower chamber includes the slot 12 in which an aerobic environment is provided. The lower chamber can hold a nutrient medium and optionally a mixed cell population including immune cells and / or fibroblasts and / or epithelial cells and / or blood cells.

[0107] During the entire modeling process, at the end, or at a specified time point, samples can be taken from either chamber (insert 4 and / or slot 12). The samples can be used for analyses such as metabolite, material transport, 16s sequencing of bacteria, RNA sequencing of cells, etc. Cell-free colon model

[0108] This model is similar to the above-mentioned human colon model and does not have representative colon cell lines (such as CACO-2 or T84). Bacteria-free colon model

[0109] This model is similar to the above-mentioned human colon model. In this model, the bioreactor is maintained in a sterile environment and not inoculated with bacteria. Airway model

[0110] Representative airway cell lines (such as Calu-3 or primary airway cultures) grow in a monolayer on the permeable support of the transwell insert 4. A cell-compatible mucus layer is placed on top of the cell layer to represent airway mucin in the model. In a specific example of this model, bacteria are optionally introduced. The apical chamber's headspace is an airspace containing gas. A nutrient medium is provided in the basolateral chamber.

[0111] In this example as a lung function model, both chambers are in an oxygenated gaseous environment. The gaseous environments in both chambers can be regulated to simulate, for example, hypercapnia / hypocapnia, hyperoxemia / hypoxemia, cigarette smoke, pollution, carbon monoxide poisoning, etc. Mucus-free airway model

[0112] This model is similar to the above-mentioned airway model. In this model, there is no mucus layer.

[0113] Those skilled in the art should understand that the above detailed examples are only for illustration and do not have any restrictive meaning. Without departing from the scope of the present invention defined by the appended claims, various variations and modifications can be made. Various modifications can be made to the examples described in detail above.

[0114] From the description of this specification and the claims, the words "comprise" and "contain" and their variants mean "include but are not limited to", and they are not intended to (and will not) exclude other parts, additives, components, integers or steps. Throughout the description and claims of this specification, unless the context requires otherwise, the singular form encompasses the plural form. In particular, when using the indefinite article, unless the context requires otherwise, the specification should be understood to cover both the plural and singular cases.

[0115] Features, integers, characteristics, compounds, chemical moieties or groups described in connection with a particular aspect, embodiment or example of the present invention are to be understood to be applicable to any other aspect, embodiment or example described herein, unless incompatible therewith. All features disclosed in this specification (including any accompanying claims, abstract and drawings), and / or all steps of any method or process so disclosed, may be combined in any manner, except combinations where at least some of such features and / or steps are mutually exclusive. The invention is not restricted to the details of any foregoing embodiments. The invention extends to any novel feature or any novel combination of features disclosed in this specification (including any accompanying claims, abstract or drawings), or to any novel step or any novel combination of steps of any method or process so disclosed.

[0116] Those skilled in the art will understand that the above embodiments are for illustrative purposes only and have no limiting significance. Various modifications and variations can be made without departing from the scope of the invention as defined by the appended claims. Various modifications can be made to the above detailed design. List of Components and Reference Numerals 2 Biological modeling device 43 Penetrable film (second) 4 Chamber (insert) 44 Catheter (first) 5 Insert arm 46 Catheter (second) 6 Bottom surface of insert 48 Penetrable film substrate 8 Inner wall of insert 50 Sheet 10 Top space of insert 52 Sheet opening (third) 12 Chamber (groove) 54 Sheet opening (fourth) 14 Bottom surface of groove 56 Protrusion 16 Inner wall of groove 58 Clamp 18 Top space of groove 60 Channel 20 Porous membrane 62 Groove 22 Seal 64 Transfer cap 23 Washer 24 Substrate 25 Air valve 26 Skirt 27 Tray 28 Flange 30 Barrier layer 32 Lid 34 Lid flange 36 Latch 38 Lid seal 40 Lid opening (first) 41 Penetrable film (first) 42 Lid opening (second)

Claims

1. A biological modeling device, comprising: A first chamber providing a first reservoir with a first gaseous environment; A second chamber providing a second reservoir with a second gaseous environment, wherein the first reservoir and the second reservoir are connected by a permeable support, and the first chamber is at least partially received into the second chamber; And A seal arranged and sealingly engaged with the first chamber and the second chamber such that the first gaseous environment of the first reservoir is separated from the second gaseous environment of the second reservoir by the seal.

2. The biological modeling device according to claim 1, wherein, The permeable support is a membrane, optionally a gas-permeable membrane.

3. The biological modeling device according to claim 1, wherein, The first chamber is configured to accommodate and hold a first biological element and / or solution, and the second chamber is configured to accommodate and hold a second biological element and / or solution.

4. The biological modeling device according to any one of claims 1 to 3, wherein, The first gaseous environment includes the headspace of the first reservoir of the first chamber, and the second gaseous environment includes the headspace of the second reservoir of the second chamber.

5. The biological modeling device according to any one of the preceding claims, wherein, The second gaseous environment is sealed from the first gaseous environment by the combination of the seal and the first chamber.

6. The biological modeling device according to any one of the preceding claims, wherein, The seal is a gasket seal.

7. The biological modeling device according to claim 6, wherein, The gasket seal includes a penetrable diaphragm, optionally a self-sealing diaphragm.

8. The biological modeling device according to claim 6 or 7, wherein, The first chamber is at least partially received into the second chamber through the gasket seal.

9. The biological modeling device according to any one of the preceding claims, wherein, The seal includes a barrier layer arranged to sealingly engage with the second chamber to separate the second gaseous environment from the first gaseous environment, the barrier layer including an opening through which the first chamber is at least partially received to form a sealed engagement between the barrier layer and the first chamber, whereby the second chamber is sealed by the barrier layer and the first chamber.

10. The biological modeling device according to any one of the preceding claims, wherein the second chamber is located in a substrate, or wherein the second chamber is formed as a part of the substrate.

11. The biological modeling device according to any one of the preceding claims, wherein, The second chamber includes a bottom surface and an inner wall extending upward from the bottom surface towards an open top, the bottom surface and the inner wall together defining the second reservoir.

12. The biological modeling device according to any one of the preceding claims, wherein, The first chamber is an insert including the permeable support.

13. The biological modeling device according to claim 12, wherein, The permeable support forms at least a part of the bottom surface of the first chamber.

14. The biological modeling device according to any one of the preceding claims, comprising a plurality of second chambers, each second chamber including a second reservoir.

15. The biological modeling device according to claim 14, comprising a plurality of first chambers, each first chamber being an insert corresponding to each of the plurality of second chambers, each insert including a first reservoir and a permeable support located between the respective first reservoir and the second reservoir.

16. The biological modeling device according to claim 15, comprising a plurality of seals, each seal being arranged between and sealingly engaging the respective first chamber and the second chamber.

17. The biomodeling device according to any one of the preceding claims, wherein, The first chamber is a top chamber, and optionally, wherein the second chamber is a base-side chamber.

18. The biological modeling device according to any one of the preceding claims, wherein, The seal is arranged between the outer surface of the first chamber and the inner wall of the second chamber and sealingly engages the outer surface of the first chamber and the inner wall of the second chamber.

19. The biomodeling device according to any of the preceding claims, further comprising a lid for the first chamber, wherein the headspace of the first chamber is enclosed by the lid.

20. The biological modeling device according to claim 19, wherein, The lid includes a seal that is arranged to sealingly engage the first chamber to separate the first gaseous environment from the surrounding ambient environment, and wherein the headspace of the first chamber is sealed by the seal.

21. The biological modeling device according to any one of the preceding claims, wherein, The seal of the lid includes at least one penetrable portion.

22. The biological modeling device according to claim 21, wherein, The seal of the lid includes at least two penetrable portions, wherein one penetrable portion is configured to allow access to one of the first chamber or the second chamber, and the other penetrable portion is configured to allow access to the other of the first chamber and the second chamber.

23. The biomodeling device according to claim 21 or 22, wherein, The penetrable portion includes a diaphragm, and optionally, wherein the diaphragm is a resealable diaphragm or a self-sealing diaphragm.

24. The biomodeling device according to any of the preceding claims is a culture device for growing cells and / or tissues in vitro.

25. A method of co-culturing cells using the biomodeling device according to any of claims 1 to 24.

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