Biomaterial testing apparatus and methods of making and using same

Abstract

A biological material testing apparatus (100) is provided that includes a substrate (102) having at least one chamber unit (104) defined therein, and a support member (106) for supporting a biological material contained in the at least one chamber unit. One of the substrate and the support member is molded on the other of the substrate and the support member. A method (10) for manufacturing such a biological material testing device is also provided. Uses and methods are also provided in which the device is used to test a biological material, for example in a drug test in which the biological material is exposed to a drug to be tested.

Description

TECHNICAL FIELD

[0001] The present invention relates to a method for manufacturing a biomaterial testing device.

[0002] The present invention also relates to the biomaterial testing device itself and to a biomaterial testing device obtainable by the method.

[0003] The present invention also relates to a mould for use in the manufacturing method.

[0004] The present invention also relates to the use of such a device for testing a biomaterial.

[0005] The present invention also relates to a drug testing method using the device. BACKGROUND

[0006] In vitro testing of mammalian cells or tissues is an important technique for obtaining clinically important information about the mammalian material under study. For example, a biopsy of mammalian cell or tissue material can be subjected to such testing to determine abnormalities or disease in such mammalian material, or to expose diseased mammalian material to a drug (e.g. an experimental drug) to monitor the response of the diseased mammalian material to such exposure. For example, this approach is frequently used in oncology procedures. This can provide important insights into how the disease in an individual can be effectively treated without having to expose the individual to a range of potentially effective drugs, which can be undesirable for a variety of reasons, including drug toxicity.

[0007] Cardiotoxicity of drugs can be tested. Many drugs adversely affect cardiac muscle contraction and profile. Drug-induced cardiotoxicity is a major side effect encountered by some clinically important drugs. This toxicity has previously led to the post-marketing withdrawal of many pharmacologically active drugs, and has limited the efficacy of other clinically useful drugs. In the past four decades, almost 10% of drugs have been recalled from global markets due to cardiovascular safety issues (see for example Mina T. Kelleni and Mahrous Abdelbasset, “Drug Induced Cardiotoxicity: Mechanism, Prevention and Management” (2018); chapters of “Cardiotoxicit” edited by Wenyong Tan).

[0008] Drug-induced cardiotoxicity thus represents a key reason for compounds to be rejected in preclinical and clinical development, which reflects the severity of cardiotoxicity as a side effect observed during the development of new drugs. Assessing the risk of drug-induced cardiotoxicity, including QT interval prolongation, is now considered an integral part of the standard preclinical evaluation of new chemical entities (cf. again “Drug Induced Cardiotoxicity: Mechanism, Prevention and Management”).

[0009] Cardiotoxicity can be tested in animal models, for example in rats. However, these models have drawbacks and are not always reliable in predicting the effect(s) experienced by humans. This can lead to potentially beneficial drug treatments being discarded, but can also mean that potentially toxic drug treatments enter human trials.

[0010] Another way of assessing cardiotoxicity is to test the effects of a drug in vitro. The drug to be tested can be added to a group of cardiomyocytes grown in vitro. Thereafter, the effects of electrical stimulation on the group of cardiomyocytes can be studied. An example of a state-of-the-art tool is “HeartDyno” (trademark) developed by the James Hudson group (cf. for example Mills et al. “Functional screening in human cardiac organoids reveals a metabolic mechanism for cardiomyocyte cell cycle arrest” (Proc Natl Acad Sci U S A. 2017; 114, 40).

[0011] A tool of this type encompassed by the term “support member” herein can comprise an oval well with two small protrusions, e.g. pillars, at the bottom. Such a support member can be produced using thin film technology. A mold can be fabricated on a wafer via SU-8 lithography, e.g. to create 700 pm deep features. Polydimethylsiloxane (PDMS) can then be cast on these features and cured. After removal of the PDMS from the wafer, 6 mm diameter samples can be punched.

[0012] The samples are for example placed in the wells of a ninety-six well substrate and glued to the bottom of the substrate using silicone glue / adhesive. A mixture of cardiomyocytes, cardiac fibroblasts, collagen, DMEM, NaOH and Matrigel can then be added to the wells and supported on the samples. It is noted that the one or more wells defined in such a substrate are referred to herein using the more general term "chamber unit". Cardiomyocyte tissue forms within a few days. The tissue shows spontaneous contractions, but also contracts during electrical stimulation. During contraction, the two posts deflect and the deflection is analyzed using a video analysis algorithm.

[0013] Fluidic devices comprising two or more fluidic channels for transporting fluids to one or more chamber units, e.g. culture chamber(s), defined in a substrate of the fluidic device are also known. For example, one channel can be used to provide nutrients and oxygen to each of the chamber unit(s) and to remove metabolic products, e.g. carbon dioxide, therefrom. A further channel can be used to provide a pharmaceutical treatment to the chamber unit(s). Such devices can comprise a support member, e.g. a multi-well support member, for supporting biological material, e.g. a cell group, a bilayer, a spheroid, an organoid or a biopsy, contained in the chamber unit(s). SUMMARY

[0014] Attaching a substrate, e.g. of the type described above, and a support member, e.g. of the type described above, to each other can present various technical challenges. It can be difficult to perform such an attachment in a sterile manner and / or without inadvertently providing a point between the substrate and the support member at which fluid leakage can occur. Furthermore, additional manufacturing steps can be required to attach the substrate and the support member to each other.

[0015] The invention is defined by the claims.

[0016] According to examples in accordance with an aspect of the present disclosure, there is provided a method for manufacturing a biological material testing device, the device comprising: a substrate, at least one chamber unit being defined in the substrate; and a support member for supporting biological material contained in the at least one chamber unit, the method comprising: molding one of the substrate and the support member on at least a portion of the other of the substrate and the support member using a mold, the at least a portion of the other of the substrate and the support member being arranged in the mold or adjacent to the mold during the molding.

[0017] By molding one of the substrate and the support member on at least part of the other one of the substrate and the support member, the former can be shaped and adhered to the latter in a single step. In this way, a separate assembly step in which the substrate and the support member are attached (e.g., adhered and / or clamped) to each other can be avoided. Adhesive / glue can not be needed to keep the substrate and the support member attached to each other. Thus, the method can provide a relatively fast and reliable process for manufacturing the device in which the substrate and the support member can be considered integrated with each other in a single piece.

[0018] Further, the support member (e.g., a silicone support member) can be anchored in the frame (e.g., a polycarbonate frame) defined by the substrate by a geometry. For example, surfaces of the substrate and the support member that will be in contact in the device can have a geometry or shape that promotes adhesion. Such a geometry or shape can be selected from the group comprising: a rough surface, corrugations, protrusions, recesses. However, other geometries can additionally or alternatively be used.

[0019] In at least some embodiments, the molding comprises injection molding the one of the substrate and the support member on the at least part of the other one of the substrate and the support member. Injection molding can provide a cost-effective and scalable way of manufacturing the device.

[0020] In some embodiments, the method comprises initially molding the substrate or the support member to form an initially molded part, and subsequently molding whichever of the substrate and the support member has not been molded on the initially molded part. In such embodiments, the method can comprise two-step injection molding, e.g., so-called “2K” injection molding.

[0021] Alternatively or additionally, molding the one of the substrate and the support member on the at least part of the other one of the substrate and the support member can comprise curing a precursor material. The precursor material is preferably liquid during its addition to the mold. In case of injection molding, the molding can comprise injecting the precursor material into the mold, and then curing the precursor material in the mold. Preferably, in this context, curing means that one or more components of the precursor material undergo a chemical or physical transformation to transform the precursor material into the solid support member material. A precursor material that undergoes a chemical transformation is preferred. The use of a precursor material facilitates the molding of a substrate and / or support member comprising a rubber material. Such materials are difficult to reshape after their formation. For example, a support member comprising a silicone polymer of the rubber type as disclosed herein is preferably made using a liquid precursor material that is cured into the silicone polymer rubber by a chemical reaction of the precursor components. Other rubber materials can also be used, and they can benefit from the use of a precursor material in the method.

[0022] Curing can include heating the precursor material, for example above 160 °C, for example between 160 °C and 200 °C. Heating at such temperatures can help ensure that the precursor material is cured and adheres to at least part of the other of the substrate and support member (e.g. the initially molded part). The heating can be used to cause the chemical and / or physical transformation to occur.

[0023] In some embodiments, initially molding the substrate or support member includes heating the material above room temperature to make the material moldable, enabling the initially molded part to be formed (e.g. heating to 200 to 300 °C in the case of a thermoplastic material (often referred to as a thermoplastic plastic)). It should be noted that the term "thermoplastic material" as used herein is intended to refer to a polymer that can be softened by heating and then processed (in particular molded).

[0024] In such embodiments, subsequently molding whichever of the substrate and support member that has not yet been molded on the initially molded part can be performed prior to the initially molded part returning to room temperature, and preferably while the initially molded part is at a temperature of at least 80 °C. This can help bond the substrate and support member to each other. In embodiments in which the subsequent molding includes curing a precursor material, such maintenance of an elevated temperature can also help such curing.

[0025] In some embodiments, the method includes arranging at least part of the base member in or adjacent to the mold, and initially molding the substrate or support member on the base member.

[0026] After molding whichever of the substrate and support member that has not yet been molded on the initially molded part, the base member, with the substrate and support member molded thereon, can be removed (e.g. released) from the mold.

[0027] The base member can help strengthen the apparatus. Alternatively or additionally, the base member (e.g. a glass base member) can be optically transparent, enabling biological material to be analyzed via optical microscopy through the glass base member.

[0028] The base member (e.g. a glass base member) can also provide a smooth surface on which to mold the support member. Such a smooth surface in turn can help cause the outer surface of the support member to have a smooth surface. This can facilitate analysis of biological material via optical microscopy through the support member.

[0029] It should be noted that in other embodiments in which a base member is not included in the apparatus (or, more generally, the outer surface of the support member is in contact with the surface of the mold during molding), the surface of the mold is preferably a polished surface.

[0030] Such a polished surface can help provide a relatively smooth outer surface of the support member, which can ultimately facilitate optical microscopic analysis of the biological material passing through the support member.

[0031] In some embodiments, the method comprises releasing the substrate together with the support member from the mold when the temperature of the substrate and the support member is higher than room temperature, preferably at least 40°C. In this way, the risk of one or both of the substrate and the support member sticking to the mold can be minimized.

[0032] In some embodiments, the method comprises adjusting the mold from a first configuration for molding the one of the substrate and the support member to a second configuration for molding the other of the substrate and the support member. In such embodiments, adjusting the mold from the first configuration to the second configuration can comprise switching or replacing the second mold part with a third mold part while the part initially molded between the first mold part and the second mold part remains housed in the first mold part. Such switching can for example comprise opening the mold by displacing the second mold part, after which the mold is reclosed using the third mold part.

[0033] In preferred embodiments, the method preferably comprises that the mold comprises a second mold configuration for molding the support member onto the substrate, the second mold configuration comprising a second mold part and a third mold part, which can be separated from the first mold part and together form the second mold configuration. In some embodiments, the substrate can have been previously molded and inserted into the first mold part before molding of the support member takes place. In other embodiments, the method comprises that the mold comprises a first mold configuration for molding the substrate, the first mold configuration comprising a first mold part and a second mold part, which can be separated from the first mold part and the substrate once molded, such that the substrate remains within the first mold part. In such embodiments, adjusting the mold from the first configuration to the second configuration can thus comprise switching or replacing the second mold part with a third mold part while the substrate initially molded between the first mold part and the second mold part remains received in the first mold part. Such switching can for example comprise opening the mold by displacing the second mold part, after which the mold is reclosed using the third mold part.

[0034] In preferred embodiments using the second mold configuration (and optionally, but preferably also using the first mold configuration), the support member is molded on a substrate. Preferably, the substrate comprises or even consists of a thermoplastic material or a thermoset material (neither of which is rubber). Preferably, such material is transparent. For example, and preferably, the material comprises at least one component selected from the group comprising polycarbonate, polyether ether ketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and polyether sulfone (PES). Most preferred material comprises polycarbonate. Such material typically has one or more of the following desirable properties: thermal properties as further described herein, transparency to visible light, rigidity (e.g., not rubber), and availability / cost. The substrate comprising or consisting of any of these materials is preferably combined with a support member comprising a rubber material. Preferred rubber material comprises a silicone, for example one or more silicone polymers as described herein. Preferably, the silicone rubber material is molded using a precursor material as described herein that undergoes curing as described herein. Most preferably, the silicone rubber material is a silicone rubber material modified with polar groups, for example carboxylic acid groups or their conjugate base groups.

[0035] In some embodiments, the support member can be formed from a material that is softer (e.g., lower Shore A hardness) and / or more flexible than the material forming the substrate. Thus, the mechanical properties of the support member can be suitable for supporting biological material, for example, cells or tissue cultured in the chamber unit(s), while the substrate contributes more to the structural rigidity of the device.

[0036] Alternatively or additionally, the support member can be formed from a material that is more biocompatible than the material forming the substrate, such that cells and / or tissue preferentially adhere to the support member. In this way, the choice of material can be used to direct cell / tissue growth, where such cell / tissue growth is expected, for example, in the area(s) of the device that facilitate observation and / or testing of the cells / tissue.

[0037] In some embodiments, the support member comprises a silicone. Preferably, the silicone comprises or even consists of a silicone polymer in the form of a rubber. Such silicone support member can be optically transparent and also have limited autofluorescence, such that the support member can allow for the use of various commonly used optical inspection techniques and protocols in tests performed using the device, for example, with or without staining. Furthermore, the silicone can be suitably biocompatible, particularly when modified such that polar groups are available at the biological material-contacting surface(s) of the silicone substrate.

[0038] More generally, the support member can comprise a polymeric material that is bulk modified with moieties each comprising a polar group, wherein the polar groups of the moieties are available at a surface of the support member that is arranged to contact the biological material housed in the at least one chamber cell. Such polar groups can help render the support member biocompatible, for example via application of proteins, such as fibronectin, to the polar group functionalized surface of the support member.

[0039] In some embodiments, the substrate is formed of a material having a glass transition temperature of at least 140 °C. This minimum glass transition temperature can allow, for example, molding of the support member on the substrate, for example, such molding including relatively high temperature curing of a precursor material.

[0040] Alternatively or additionally, the substrate can be formed of a thermoplastic, preferably a thermoplastic selected from one or more of the following: polycarbonate, polyether ether ketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and polyethersulfone (PES).

[0041] In some embodiments, the substrate provides a side wall(s) of the at least one chamber cell, wherein a central portion of the support member is provided in the at least one chamber cell, and wherein a plurality of rib elements each extend from the central portion to engage with the side wall of the respective chamber cell. Such rib elements can help minimize deformation of the central portion while keeping shear stress sufficiently high to help hold the support member in place.

[0042] In some embodiments, the support member comprises at least one structural feature for contacting the biological material housed in the at least one chamber cell. Such structural feature(s) can provide a suitable shape or profile for growth of cells or tissue thereon. The shape or profile of the structural feature(s) can be selected depending on, for example, the type of cells or tissue being grown and / or the test to be performed on the biological material. The structural feature can be selected from the group comprising: a membrane; a porous membrane; a plurality of pores each having a membrane or porous membrane and a membrane having an opening or slit for clamping a biological material such as a biopsy. However, other structural features can be selected.

[0043] In some embodiments, the at least one structural feature of each chamber cell comprises a pair of flexible protrusions, for example flexible struts or posts, for supporting tissue thereon and therebetween. In such embodiments, the apparatus can be used to test muscle tissue, for example heart tissue, grown between and around the pair of protrusions. When the muscle tissue contracts, the pair of protrusions can deflect, wherein the displacement of the protrusions caused by the deflection is measurable to enable determination of the force provided by the contraction of the tissue.

[0044] In some embodiments, the at least one chamber unit comprises a plurality of chamber units. Thus, testing of biological material can be performed simultaneously in different chamber units. In such embodiments, the chamber units can be arranged, for example, in one or more rows in a manner similar to conventional well plates.

[0045] According to another aspect, there is provided a mold system for molding a biological testing device using a method as disclosed herein. The mold system comprises a first mold part and a third mold part separable from the first mold part, wherein the first mold part and the third mold part are designed and arranged to be combined to form a second mold configuration, the second mold configuration being arranged to hold one of a substrate and a support member, such that in the second mold configuration the first mold part, the second mold part, and the one of the substrate and the support member enclose one or more second open spaces, wherein the other of the substrate and the support member can be molded onto the one of the substrate and the support member. In preferred embodiments, one of the substrate and the support member consists of a substrate, while the other of the substrate and the support member consists of a support member. In such embodiments, the second mold configuration is configured for molding a support member onto a substrate.

[0046] Preferably, the first mold part comprises at least one first mold surface for defining a chamber surface of the end portion. Thus, the first mold surface can be used to design the end portion surface that will eventually be used to support biological material. Preferably, the first mold part comprises or consists of a chamber protrusion comprising the first mold surface, wherein the chamber protrusion extends within the mold space of the second mold configuration, and wherein, if the mold configuration holds a substrate, the protrusion occupies (e.g. fills in) at least part of the chamber of the substrate, such that the first mold surface is exposed in the one or more second open spaces of the second mold configuration. Thus, the first mold surface can be shaped to provide a desired design for the end portion of the support member, such as a post, a membrane, a well, etc. as defined herein. The first mold surface can have any shape as desired as described herein, such as flat, curved, with protrusions or indentations or recesses according to the desired shape of the support member to be formed against the surface.

[0047] These mold designs can be used to manufacture chamber units, some of the walls of which are defined by a substrate and some of the walls of which are defined by a support member, such that in the chamber at least some surfaces of the support member are exposed to support biological material.

[0048] The first mould part is further arranged such that, if it holds the substrate, at least one (preferably multiple) contact surface of the substrate is exposed within one or more spaces of the second mould configuration. Thus, the support member can be moulded onto these contact surfaces using the second mould configuration.

[0049] The first mould part can comprise one or more protrusions for occupying (e.g. filling) the channels in the substrate.

[0050] Preferably, the third mould part comprises at least one third mould surface for defining the end portion. Thus, the first mould surface can be used to design the end portion surface that will be used to support the biological material in the finished device. The third mould surface can be arranged to define one or more of the following: a flange, a membrane or a pillar structure. The first mould surface and the third mould surface are preferably arranged in the respective mould parts such that, in the second mould configuration, they define parts of the one or more second open spaces in which the end portion of the support member is defined. Preferably, the third mould surface is comprised in a protrusion that is arranged in the third mould part such that it extends towards the first mould surface in the one or more open second openings of the second mould configuration. Preferably, the protrusion defines a sub-chamber in the support member. Preferably, the third mould part comprises one or more channel protrusions for forming one or more channels in the support member. For example, some of these protrusions are arranged to form channels that are connected to one or more sub-chambers.

[0051] Preferably, the mould system further comprises a second mould part that is separable from the first mould part and designed and arranged to combine with the first mould part to form a first mould configuration, wherein the first mould part and the second mould part enclose one or more first open spaces in which one of the substrate and support member can be moulded. This second mould part is not needed in the method using only the second mould configuration. For example, it is useful and advantageous in the method comprising moulding of the substrate when a pre-fabricated substrate is used in the process.

[0052] Preferably, the second mould part comprises a second mould surface for defining a contact surface of the substrate to be moulded. It is this second mould surface that can be shaped to improve the adhesion of the support member to the substrate. Preferably, the second contact surface comprises at least one of the following: corrugations, indentations, protrusions and surface roughness. Any shape that increases the area of the contact surface relative to the area of a flat contact surface is beneficial to adhesion.

[0053] Preferably, the second mould part comprises a further chamber protrusion which is arranged to extend in the one or more first open spaces of the first mould configuration, such that in the first mould configuration the combination of the chamber protrusion of the first mould part and the further chamber protrusion define a chamber cell in the substrate.

[0054] The mould system is preferably a mould system for injection moulding. Preferably, one or more of the mould parts comprise one or more openings for injecting material to be moulded into the substrate and the support member. Preferably, the mould parts are made of metal, such as steel.

[0055] Preferably, the first, second and third mould parts are shaped and dimensioned such that the apparatus (and preferably the substrate of such apparatus) comprises at least one (and preferably a plurality of) chambers.

[0056] According to another aspect, there is provided an apparatus for testing a biological material, comprising: a substrate defining at least one chamber cell therein; and a support member for supporting a biological material housed in the at least one chamber cell, one of the substrate and the support member being moulded on the other of the substrate and the support member.

[0057] The apparatus can be obtainable by a method according to any of the embodiments described herein. The apparatus can be obtainable by moulding one of the substrate and the support member on the other of the substrate and the support member.

[0058] The apparatus preferably comprises at least one chamber, wherein the at least one chamber is defined by a portion of a wall of the chamber cell and a portion of a surface of the support member. The portion of the support member preferably comprises or consists of an end portion, such as those defined herein. Preferably, the end portion is arranged in the chamber cell such that it forms a bottom of the chamber or at least a part of a bottom of the chamber.

[0059] The chamber unit can have one chamber or multiple chambers. The device can have multiple chambers. These can be arranged in one or more arrays. In some embodiments, the device comprises fluidic channels from and to the chambers. Preferably, these channels are arranged in the base plate. In some embodiments, the chambers are arranged to connect multiple chambers. Alternatively or additionally, the support member comprises at least one, such as multiple, fluidic channels. In some embodiments, the device comprises at least one membrane, wherein such membrane is part of the support member. Preferably, the membrane is part of the end portion. In some embodiments, such membrane is porous. In some embodiments, the end portion comprises at least one aperture. In some embodiments, the aperture comprises a membrane. The end portion can comprise a flange extending at least partially into the chamber unit. The flange is arranged to carry or be integrated with a membrane or part of the end portion comprising an aperture as defined herein. Preferably, the end portion comprises a porous membrane and a flange, and the support member is further arranged to define a sub-chamber, which is delimited by the membrane and connected to one or more fluidic channels arranged in the support member.

[0060] Embodiments described herein with respect to the method can be applicable to the device, and embodiments described herein with respect to the device can be applicable to the method. Thus, preferred materials for the method are also preferred materials for the device itself. Similarly, preferred structural device features of the device described with respect to the method disclosed and defined herein are also preferred features for the device itself.

[0061] According to yet another aspect, there is provided use of a device according to any of the embodiments described herein for testing biological material.

[0062] According to another aspect, there is provided a method of drug testing, comprising providing biological material in at least one chamber unit of a device according to any of the embodiments described herein, and exposing the biological material to a drug to be tested.

[0063] In some embodiments, providing biological material in the at least one chamber unit comprises culturing cells in the at least one chamber unit.

[0064] Alternatively or additionally, the method of drug testing can comprise monitoring the response of the biological material, e.g. cultured cells, to the drug to be tested. This monitoring can be carried out in any suitable manner, e.g. by optical microscopy, e.g. through the support member when the support member is optically transparent.

[0065] These and other aspects of the present disclosure will be apparent from and elucidated with reference to the embodiments(s) described hereinafter. BRIEF DESCRIPTION OF DRAWINGS

[0066] For a better understanding of the present disclosure, and to show more clearly how it can be carried into effect, reference will now be made, by way of example only, to the accompanying drawings in which:

[0067] FIG. 1A and FIG. 1B A method for manufacturing a biomaterial testing device according to a first example is schematically depicted;

[0068] FIG. 2A Portions of an exemplary support member for inclusion in a device are shown;

[0069] FIG. 2B Views of another exemplary support member for inclusion in a device are provided;

[0070] FIG. 3 A flowchart of a method for manufacturing a biomaterial testing device according to another example is provided;

[0071] FIG. 4 A device according to a second example is shown;

[0072] FIG. 5A to FIG. 5C Various views of a device according to a third example are provided;

[0073] FIG. 6 A device according to a fourth example is shown;

[0074] FIG. 7 A device according to a fifth example is shown; and

[0075] FIG. 8 A device according to a sixth example is shown.

[0076] FIG. 9A A set of mold parts for forming a first mold configuration of a mold to be used for molding a substrate of a device of FIG. 9E to FIG. 9H using the methods disclosed herein is shown;

[0077] FIG. 9B A first mold configuration of mold parts is shown that is used to mold a substrate of a device of FIG. 9A wherein; FIG. 9E to 9H the substrate of the device is molded;

[0078] FIG. 9C and FIG. 9D A second mold configuration of mold parts is shown that is used to replace mold parts to provide a support member for molding a device of FIG. 9E to FIG. 9H using the methods disclosed herein. The second mold configuration and the support member molded therein are shown in FIG. 9D

[0079] FIG. 9E to 9H ​Cross-sections of devices molded according to the methods disclosed herein are shown, as well as top and bottom views. DETAILED DESCRIPTION

[0080] The present disclosure will be described with reference to the accompanying drawings.

[0081] The detailed description and specific examples, while indicating exemplary embodiments of the devices, systems, and methods, are intended for purposes of illustration only and are not intended to limit the scope of the present disclosure. These and other features, aspects, and advantages of the devices, systems, and methods of the present disclosure will become better understood from the following description, appended claims, and accompanying drawings. The drawings, which are not necessarily to scale, depict embodiments of the present disclosure. It should be noted that the detailed description and specific examples are intended to illustrate, not limit, the scope of the present disclosure. Furthermore, the figures are not drawn to scale, and the dimensions of the various features are not necessarily to scale.

[0082] A biomaterial testing device is provided, comprising a substrate defining at least one chamber unit therein, and a support member for supporting a biomaterial accommodated in the at least one chamber unit. One of the substrate and the support member is molded on the other of the substrate and the support member. A method for manufacturing such a biomaterial testing device is also provided. Uses and methods are also provided in which the device is used for testing a biomaterial, e.g. in a drug test, wherein the biomaterial is exposed to a drug to be tested.

[0083] FIG. 1A and FIG. 1B An apparatus design and manufacturing method 10 according to examples in accordance with the present disclosure is schematically depicted. The method 10 is for manufacturing an apparatus 100 for testing a biomaterial. To this end, the apparatus 100 comprises a substrate 102 defining at least one chamber unit 104 therein. FIG. 1A and FIG. 1B The substrate 102 shown has a plurality (in this case 8) of such chamber units 104. The apparatus 100 further comprises a support member 106 for supporting a biomaterial when accommodated in each of the chamber unit(s) 104 (the biomaterial being accommodated in the chamber unit(s) 104 in the FIG. 1A and FIG. 1B are not visible in the figure).

[0084] Examples of biomaterials include cell panels, bilayer cells, spheroids, organoids, or biopsies, but other biomaterials can also be used.

[0085] Each chamber unit 104 can have a height extending between a top side and a bottom side of the substrate 102 in the range of 1.8 to 2.6 mm, e.g. about 2.2 mm. Alternatively or additionally, a diameter of each chamber unit 104 can be in the range of 3 to 20 mm, e.g. in the range of 3 to 10 mm, e.g. about 6 mm.

[0086] Such a scale can balance the following two requirements: the requirement to keep the (one or more) chamber units 104 as small as possible (e.g., so that as many (one or more) chamber units 104 as possible can be defined in the substrate 102); and the requirement to provide sufficient space for the (one or more) chamber units 104 to test biological materials (e.g. for capturing and subsequently analyzing cells).

[0087] One or more chamber units 104 can have any suitable shape. From a fluid dynamics perspective, such as FIG. 1A and FIG. 1B The generally cylindrical shape shown may be preferred, for example, to mitigate the risk of fluid pooling at the corners of chamber unit 104 and / or to ensure availability with respect to existing analytical equipment. However, shapes other than cylindrical are contemplated, such as those with square or rectangular cross-sections.

[0088] In some embodiments, such as in FIG. 1A and FIG. 1B In the illustrated embodiment, substrate 102 provides at least one or more sidewalls 108 of chamber unit 104, such as one or more cylindrical sidewalls 108, and support member 106 provides end portions 110 from which the sidewalls 108 extend. The end portions 110, combined with the sidewalls 108 of each chamber unit 104, can provide a container in which biological material can reside, for example, along with cell culture medium. Typically, the end portion 110 may be referred to as the bottom of a chamber, and the chamber may also be referred to as a container.

[0089] Typically, each chamber or container has a chamber depth measured along the sidewalls from the top side of the substrate to the end portion. FIG. 1A to FIG. 1B In the example, the height equals the chamber depth, but this is not always the case. In a variation of the example, as will be disclosed herein, the support member can extend from the bottom side of the chamber unit into the chamber unit such that the chamber depth is less than the distance the support member extends into the chamber unit to form the bottom side.

[0090] The thickness of the support member 106 (e.g., one or more end portions 110 provided by the support member 106) can be selected to enable analysis of the biomaterial contained in the chamber unit 104 through the support member 106.

[0091] Such analysis can include optical microscopy with or without staining. To this end, the support member 106, e.g. at least the end portion(s) 110 provided by the support member 106, can be optically transparent and can have a thickness of e.g. at most 0.4 mm. In this sense, optical can relate to spectral regions including one, more or all of visible light, UV light and near infrared light.

[0092] In at least some embodiments, the at least one chamber unit 104 comprises a plurality of chamber units 104. There is then a plurality of chamber units in the substrate and thus also in the apparatus. Accordingly, testing of biological material can be performed in different chamber units 104 simultaneously. In such embodiments, the chamber units 104 can be arranged in one or more rows 105. In FIG. 1A and FIG. 1B One such row 105 comprising eight chamber units 104 is shown in

[0093] In some embodiments, e.g. in the embodiment shown in FIG. 1A and FIG. 1B The apparatus 100, e.g. the substrate 102 thereof, comprises one or more mounting features 111, such as tabs, to enable the apparatus 100 to be mounted on and / or in a holder (not visible). In such embodiments, the holder can for example be configured to support a plurality of such apparatuses 100 simultaneously thereon and / or therein.

[0094] The holder can for example hold FIG. 1A and FIG. 1B twelve of the apparatuses 100 shown in

[0095] In some embodiments, one or more of the apparatuses 100 can be individually detachable from the holder.

[0096] This can provide for a more flexible arrangement of the holder apparatus(es). In relation to the non-limiting example of twelve of the apparatuses 100 being mountable in the holder, detachment of one or more of the apparatuses 100 can be performed when the experimenter does not wish to use all ninety-six chamber units 104 at once.

[0097] During at least part of the test performed using the device 100, it is often desirable for the biological material to survive. Therefore, the device 100, e.g. at least its support member 106, can be formed from a suitable biocompatible material. In some embodiments, the support member 106 is formed from a biocompatible polymeric material, such as silicone, to which, e.g., a suitable cell culture protein, such as fibronectin, can or is applied. Suitable materials are mentioned herein.

[0098] In some embodiments, the support member 106 comprises a polymeric material that is bulk-modified with motifs each comprising a polar group, wherein the polar groups of the motifs are available at a surface of the support member 106 that is arranged to contact the biological material accommodated in the at least one chamber unit 104.

[0099] Such polar groups can help to render the support member 106 biocompatible, e.g. via functionalization of the polar group-functionalized surface of the support member with a protein, such as fibronectin.

[0100] In some embodiments, the motifs comprise fatty acid motifs, wherein the polar group comprises a carboxylic acid group of the fatty acid motif.

[0101] Particularly mentioned are silicones that are bulk-modified with motifs comprising polar groups, e.g. fatty acids, e.g. via crosslinking between silicon-hydrogen bonds and C=C bonds of unsaturated fatty acid motifs.

[0102] Such crosslinking can be catalyzed by a platinum catalyst.

[0103] In this regard, reference is made to the materials and methods described, e.g., in WO2021058657 and WO2019015988, which also describe the processing of preferred materials to be used for the manufacture of support members as defined herein, and the entire contents of which are incorporated herein by reference. Thus, in some embodiments, the one or more acidic groups are phosphorus-based acidic groups, sulfur-based acidic groups, and carboxylic acid groups or a mixture of two or more of these. Phosphoric acid groups and their conjugate bases are preferred over sulfonic acid groups, and carboxylic acid groups are preferred over phosphoric acid groups. Residues carrying such groups are more readily available during bulk modification, as their precursors have acidic groups in conjugate base form with a metal counterion during such modification processes, and such residues mix better with other constituent ingredients to form elastomers therefrom. The acidic groups can be selected according to the pKa required (as specified previously herein).

[0104] In some preferred embodiments, the residue comprises or consists of an aliphatic moiety comprising 3 or more carbon atoms and less than 50 carbon atoms, the one or more acidic groups are covalently coupled to the aliphatic moiety, and the aliphatic moiety is covalently coupled to the elastomer bulk. The aliphatic moiety can be linear or branched. It can comprise one or more carbon-carbon double or triple bonds or aromatic or benzene units. The aliphatic moiety can comprise cyclic units, such as cyclohexyl or cyclopentyl or others. The aliphatic moiety is preferably a saturated hydrocarbon moiety. The aliphatic moiety preferably comprises only carbon and hydrogen atoms.

[0105] In some embodiments, the aliphatic moiety is a linear chain coupled at the end to the elastomer bulk. In some embodiments, the aliphatic moiety does not comprise carbon-carbon triple bonds to increase the flexibility of the residue.

[0106] In some embodiments, the residues within the bulk-modified elastomer can differ from each other, each being selected as defined herein. In some embodiments, at least some of the residues can be bound to the elastomer bulk via two covalent bonds. This can for example result from a residue precursor having two ethylenic bonds for participating in the modification reaction. In some embodiments, the moiety comprises at least 5 carbon atoms, and more preferably at least 10 carbon atoms. Preferably, the aliphatic moiety comprises less than 40 or less than 30 carbon atoms. Linear moieties are preferred, but this is not a requirement. The number of carbon atoms of such moieties is preferably between 5 and 30, more preferably between 5 and 20 or between 5 and 15. The moiety and / or chain can have one or more aromatic groups. One or more of the acidic groups can be attached directly to an aromatic group, such as a benzene ring.

[0107] In some embodiments, the residue is the remainder of an unsaturated fatty acid precursor, covalently bound to the elastomer bulk via reaction of one or more of their ethylenic groups (carbon-carbon double bonds), if present.

[0108] For example, the unsaturated fatty acid residue is one or more residues of a fatty acid selected from the group comprising myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linolenic acid, a-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, and docosahexaenoic acid

[0109] In some embodiments, the residue is the remainder of a precursor having a linear or branched alkyl chain carrying one or more acidic groups and at least one ethylenic group (carbon-carbon double bond). Preferably, at least one such ethylenic group is a terminal chain ethylenic group. Terminal ethylenic groups can provide increased reactivity compared to non-terminal ethylenic groups during the formation of the elastomeric body during the manufacturing process. A linear chain having one terminal ethylenic group and one acidic group (e.g., a hydroxy acid group) is a preferred example. In such cases, there can be 5 to 15 carbon atoms in the chain.

[0110] In some embodiments, the elastomeric body comprises a silicone or a polydiene backbone. Polybutadiene and polyisoprene are examples of polydienes. Polydimethylsiloxane is an example of a polysiloxane (silicone). Silicones have higher water permeability and are more transparent, allowing for easier optical inspection of cell culture.

[0111] In some embodiments, the residue is covalently bound to the elastomeric body as a result of a reaction between the unsaturated carbon-carbon bond of the precursor of the residue and an ethylenic or hydride functional group of the elastomeric body. In polydienes, such a reaction is an ethylenic group, while in silicones, such a binding is typically with a silyl hydride functional group.

[0112] More generally, as used herein, the term "biocompatible" can refer to a material that is capable of allowing cells to proliferate on the material.

[0113] The term "more biocompatible material" can refer to a material on which cells proliferate to a greater extent than a given material being compared to the more biocompatible material.

[0114] In some embodiments, the support member 106 is formed of a material that is more biocompatible than the material forming the substrate 102, such that cells and / or tissue preferentially adhere to the support member 106.

[0115] For example, a silicone (e.g., a silicone modified in the manner described above) can form the support member 106, while a polymeric material to which biological materials have a lower affinity (e.g., a polymeric material including one or more materials selected from the group consisting of polycarbonate, polyether ether ketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and polyethersulfone (PES)) can form at least a portion of the substrate 102. For example, a polymeric material to which biological materials have a lower affinity (e.g., polycarbonate, polyether ether ketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and / or polyethersulfone (PES)) can form the sidewall(s) 108 of the chamber unit(s) 104.

[0116] In at least some embodiments, the support member 106 comprises an elastomeric material (sometimes also referred to herein as rubber or rubber material), such as an elastomeric material comprising one or more materials selected from the group consisting of silicone and polybutadiene. The elastomeric properties of such elastomeric materials (e.g. together with a biocompatible or at least modifiable to provide sufficient biocompatibility such material) can make such elastomeric materials particularly suitable for inclusion in the support member 106.

[0117] Alternatively or additionally, the support member 106 can be formed from a material that is softer (e.g. lower Shore A hardness) and / or more flexible than the material forming the substrate 102. Thus, the mechanical properties of the support member 106 can be adapted to support biological material (e.g. cells or tissue being cultured in the chamber unit(s) 104), whereas the substrate 102 contributes more to the structural rigidity of the apparatus 100.

[0118] It is also noted that embodiments in which the support member 106 comprises (e.g. is formed from) silicone, the silicone support member 106 can be optically transparent and also has limited autofluorescence, such that the support member 106 can allow the use of various commonly used optical inspection techniques and protocols in tests performed using the apparatus 100, e.g. with or without staining.

[0119] The selection of materials for the substrate 102 and the support member 106 is further discussed below in more detail in relation to a method 10 of manufacturing the apparatus 100.

[0120] A list of example materials for the support member 106 is provided below.

[0121] In some embodiments, and again with reference to FIG. 1A and FIG. 1B the support member 106 comprises at least one structural feature 112 for contacting biological material housed in the chamber unit(s) 104. Such a structural feature 112 can provide a suitable shape or profile for the growth of cells or tissue thereon. The shape or profile of the structural feature 112 can be selected according to, for example, the type of cells or tissue being grown and / or the test to be performed on the biological material.

[0122] In some embodiments, for example in FIG. 1A , FIG. 1B , FIG. 2A and FIG. 2BIn the illustrated embodiment, the at least one structural feature 112 comprises, for each chamber cell 104, a pair of flexible protrusions, e.g. flexible columns or struts, for supporting tissue thereon and therebetween. In such embodiments, the apparatus 100 can be used to test muscle tissue, e.g. cardiac tissue, grown between and around the pair of protrusions. When the muscle tissue contracts, the pair of protrusions can deflect to bend towards each other, wherein the displacement of the protrusions caused by the deflection is measurable to enable determination of one or more contractility properties, such as, for example, the extent of contraction, the frequency of contraction and / or the force of contraction provided by the contraction of the tissue.

[0123] Such measurements are described, for example, in WO 2022 / 112291, which article is incorporated herein in its entirety by reference.

[0124] The protrusions can have any suitable height, as long as they are able to fulfil their tissue testing function. In some embodiments, and with reference to FIG. 2B , the protrusions each have a height H or maximum dimension of 500 pm to 1500 pm, preferably 600 pm to 1000 pm.

[0125] Such height H (e.g. together with the thickness of the support member 106 between the outer surface of the support member 106 to the inner surface 116 from which the protrusions protrude, which is at most 4 mm) can help to analyse the tissue by optical microscopy through the support member 106 when the support member 106 is optically transparent at least underneath the protrusions.

[0126] Alternatively or additionally, the protrusions can each have a width W of 100 pm to 300 pm and / or a length L of 400 pm to 600 pm.

[0127] The spacing SP between the protrusions (e.g. protrusions having the above-mentioned height H, width W and / or length L dimensions) can be 800 pm to 1200 pm.

[0128] It is noted that the flexibility of the protrusions can be provided at least by the protrusions of the support member 106 comprising an elastomeric material, such as an elastomeric material comprising one or more materials selected from the group consisting of silicone and polybutadiene.

[0129] In some embodiments, and as best shown in FIG. 2A and FIG. 2B A recess 114 defining each chamber cell 104 is defined in the support member 106. The base surface 116 partially delimiting the recess 114 can be connected to the main surface 118 of the support member 106 by a side surface 120 extending between the base surface 116 and the main surface 118.

[0130] The structural feature(s) 112 (e.g., flexible protrusions) can be arranged on and / or in the base surface 116 of the recess 114. Alternatively or additionally, the recess 114 can be elongate, e.g., elliptical, when viewed in plan, as shown in the upper pane of FIG. 1B and FIG. 2B .

[0131] In the case of such an elongate recess 114, and when protrusions are included in the support member 106, the protrusions can be arranged along a longitudinal axis 122 along which the recess 114 is elongate. Contraction of the muscle tissue between the protrusions can be along this longitudinal axis 122.

[0132] The recess 114 (e.g., elongate recess 114) can have any suitable dimensions. In some embodiments, and with reference to FIG. 2B , the recess 114 has a width W1 of 1500 to 2500 pm, preferably 1800 to 2200 pm, and / or a length L1 of 2500 to 3500 pm, preferably 2700 to 3200 pm. In embodiments in which the recess 114 is elongate, the length L1 extending along the longitudinal axis 122 is significantly greater than the width W1.

[0133] The height H1 of the recess 114 (e.g., recess 114 having the above-mentioned width W1 and / or length L1 dimensions) can be 500 pm to 1500 pm, preferably 600 pm to 1000 pm.

[0134] It is noted that the height H1 of the recess 114 can be the same as or within 10% of the height H of the protrusions (when present).

[0135] It is noted that in some embodiments, the structural feature(s) 112 (e.g., protrusions) are included in the support member 106, but no recess 114 is defined in the support member 106.

[0136] In such embodiments, suitable lateral confinement of the biological material, cell media, etc. within each of the chamber(s) unit(s) 104 can rely on the side wall(s) 108 of the chamber(s) unit(s) 104.

[0137] In some embodiments, e.g., in the embodiment shown in FIG. 2A , a central portion 124 of the support member 106 is provided in, e.g., each of the chamber(s) unit(s) 104, wherein a plurality of rib elements 126 each extend from the central portion 124 to engage the side wall 108 of the respective chamber unit 104. Such rib elements 126 can help to reduce or minimize deformation of the central portion 124 while keeping the shear stress high enough to help hold the support member 106 in place.

[0138] Any suitable number of rib elements 126 can be envisaged, such as two, three, four (as shown), five or more. Preferably, a symmetrical arrangement of rib elements 126 is employed, wherein two rib elements 126 are arranged along a first axis 128, and two rib elements 126 are arranged along a second axis 130 perpendicular to the first axis 128. FIG. 2A

[0139] It is noted that the recess 114 can be defined in the central portion 124 of the support member 106, e.g. wherein the structural feature(s) 112 (e.g. flexible protrusions) are arranged on and / or in the base surface 116 of the recess 114. In case of an elongated recess 114, the above-mentioned longitudinal axis 122 can coincide with the axis 128 along which the rib elements 126 extend to reach the side wall 108 of the chamber unit 104.

[0140] In some embodiments, the support member 106 is porous so as to render the support member 106 fluid-permeable, e.g. gas-permeable. Such a porous support member 106 can be obtained in any suitable manner, e.g. by forming pores in the material (e.g. elastomeric material) constituting the support member 106. Such pores can be formed, e.g. by laser ablation or cast-embossed poration.

[0141] Laser poration / ablation is described, e.g. in US2014127744 and US20180315409. Cast-embossed poration is described, e.g. in US2020360923, US2015010919 and US2022228108, all of which are incorporated herein by reference in their entirety.

[0142] Each of the pores is preferably dimensioned to limit or prevent cells supported on the support member 106 from passing therethrough. To this end, the diameter of each pore can be less than 10 pm.

[0143] Generally, a chamber unit as disclosed herein (e.g. as referenced to FIG. 1A to Fig. 1D and FIG. 2A and FIG. 2B ​Attaching the different materials of the substrate 102 and the support member 106 of the described apparatus to each other can present various technical challenges. It can be difficult to perform such attachment in a sterile manner and / or to perform such attachment without inadvertently providing a point between the substrate 102 and the support member 106 at which fluid leakage can occur, thereby potentially impeding use. Moreover, additional manufacturing steps can be required to attach the substrate 102 and the support member 106 to each other, thereby increasing manufacturing complexity and cost. Furthermore, in embodiments in which the support member 106 includes flexible protrusions, the substrate 102 and the support member 106 can need to be aligned to ensure that the protrusions are properly oriented, e.g., in each of the plurality of chamber units 108.

[0144] For at least these reasons, and with reference again to FIG. 1A and FIG. 1B , the method 10 according to the present disclosure includes molding 12 one of the substrate and the support member 102, 106 onto at least a portion of the other of the substrate and the support member 106, 102, the at least a portion of the other of the substrate and the support member 106, 102 being arranged in or adjacent to a mold during the molding 12. For example, in the embodiment schematically depicted in FIG. 1A and FIG. 1B , the support member 106 is molded onto the substrate 102.

[0145] By molding 12 one of the substrate and the support member 102, 106 onto at least a portion of the other of the substrate and the support member 106, 102, the former can be simultaneously shaped and adhered to the latter. In this way, a separate assembly step in which the substrate 102 and the support member 106 are attached (e.g., adhered) to each other can be avoided. In particular, an adhesive / gel can not be required to hold the substrate 102 and the support member 106 attached to each other.

[0146] The method 10 can provide a relatively quick and reliable process for manufacturing the apparatus 100, in which the substrate 102 and the support member 106 can be considered to be integrated with each other in a single piece. Moreover, the support member 106 (e.g., a silicone support member 106) can be anchored by geometry into a frame (e.g., a polycarbonate frame) defined by the substrate 102. Thus, portions of the substrate can be embedded in portions of the support member, such that one cannot be easily separated from the other.

[0147] Accordingly, the adhesion (e.g., leak-proof adhesion) provided by molding 12 one of the substrate and support member 102, 106 on the other can be supplemented by geometric anchoring, particularly in embodiments in which the support member 106 includes a central portion 124 and rib elements 126 that engage the sidewall(s) 108 of the chamber unit(s) 104. In other embodiments, the central portion-rib element arrangement 124, 126 can be omitted, e.g., to simplify the design of the mold.

[0148] It is also noted that, at a microscopic scale, irregularities (e.g., molecular-scale induced irregularities) of the surface of the first molded part (e.g., the substrate 102) can be at least partially followed by the second molded part (e.g., the support member 106) (as a result of the second molded part being molded onto the first molded part). This can increase the adhesion of the two parts of the device disclosed herein that has been manufactured using the methods disclosed herein, and benefit its leak characteristics. Accordingly, the method of molding one part onto another can allow for the use of irregularly shaped contact surfaces between the substrate and support member that would otherwise be more difficult to achieve good contact. For example, individually manufactured components with complementary and matching complex irregular shapes for close contact can be difficult to manufacture or more expensive to manufacture. Accordingly, an inspection of the contact surface between the substrate and support member of a device as disclosed herein can be used to distinguish a device as disclosed herein from other multi-component devices that have not been made using the methods as disclosed herein (e.g., methods that utilize clamping, adhering, or gluing of separately manufactured parts). Although conformal adhesion can also be observed for parts that are glued together, the additional layer of glue here can help to distinguish such glued devices from glued devices that have been manufactured using the methods disclosed herein.

[0149] Implicit in the molding 12 is that one of the substrate and support member 102, 106 (on which the other of the substrate and support member 106, 102 is molded) can withstand the molding conditions. This can affect one or more of: which of the substrate 102 and support member 106 is molded 12 onto the other (and, conversely, which component has been manufactured prior to the molding 12); the materials and / or precursors selected for forming the substrate 102 and support member 106; and the conditions during the molding 12.

[0150] In at least some embodiments, the molding 12 includes injection molding one of the substrate and support member 102, 106 on (e.g., defined by) at least a portion of the other of the substrate and support member 106, 102. Injection molding can provide a cost-effective and scalable way of manufacturing the device 100.

[0151] In some embodiments, e.g., inFIG. 3 In the embodiment illustrated in FIG. 1, the method 10 includes releasing 14 the substrate 102 and the support member 106 from the mold while the temperature of the substrate 102 and the support member 106 is higher than room temperature, preferably at least 40°C, for example between 40 and 80°C. In this way, the risk of one or both of the substrate 102 and the support member 106 sticking to the mold can be minimized.

[0152] In some embodiments, and still referring to FIG. 3 , the molding 12 (e.g., injection molding) of one of the substrate and support member 102, 106 on at least a portion of the other of the substrate and support member 106, 102 includes curing 12B the precursor material. In the case of injection molding, the molding 12 can include injecting 12A the precursor material into the mold and then curing 12B the precursor material in the mold and on the other of the substrate and support member 106, 102.

[0153] In such embodiments, the curing 12B can include heating the precursor material, for example above 160°C, for example between 160°C and 200°C.

[0154] Heating at such temperatures can help ensure that the precursor material is cured and adheres to the initially molded part.

[0155] After the heating, the substrate 102 and the support member 106 can be cooled to room temperature, although the substrate 102 is preferably released 14 from the mold before the substrate 102 and the support member 106 are cooled to room temperature, as previously described.

[0156] It is noted that the thermal curing 12B of the precursor material can require that a suitable thermally robust material be selected for one of the substrate and support member 102, 106 on which the other of the substrate and support member 106, 102 is molded via the curing 12B.

[0157] To this end, the glass transition temperature of the material from which one of the substrate and support member 102, 106 (on which the other of the substrate and support member 102, 106 is molded) can be at least 140°C.

[0158] This minimum glass transition temperature can allow for a relatively high temperature curing 12B of the precursor material during the molding 12.

[0159] Examples of materials having a glass transition temperature of at least 140°C include: polycarbonate, polyether ether ketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), polyethersulfone (PES), and combinations of the above polymers, for example a blend or alloy of at least two of polyether ether ketone (PEEK), polyetherimide (PEI), and polyethersulfone (PES).

[0160] In embodiments in which the support member 106 comprises or is formed from silicone, the curing 12B precursor material can comprise a curing silicone precursor material.

[0161] Such curing 12B can for example be carried out between 140°C and 180°C. Alternatively or additionally, the precursor material (e.g. silicone precursor material) can be injected into the mould under a pressure between 100 bar and 600 bar, preferably when the mould is above 110°C.

[0162] In some embodiments, for example in the embodiments shown in FIG. 1A , FIG. 1B and FIG. 3 , the method 10 comprises initially moulding 16 (e.g. initially injection moulding 16) the substrate 102 or the support member 106, and subsequently moulding 12; 12A, 12B whichever of the substrate 102 and the support member 106 has not been moulded on the initially moulded part (e.g. subsequently injection moulding 12; 12A, 12B whichever of the substrate 102 and the support member 106 has not been moulded on the initially moulded part). Thus, the method 10 can comprise a two-step moulding process, for example a two-step injection moulding process.

[0163] The initial moulding 16 can involve injection moulding 16 of a thermoplastic, for example a thermoplastic selected from one or more of the following: polycarbonate, polyether ether ketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI) and polyethersulfone (PES). Such a thermoplastic can for example be melted at a temperature of 200 to 300°C, injected into a mould at a pressure between 600 and 2000 bar, the mould preferably being at a temperature lower than the glass transition temperature of the thermoplastic. For example, the temperature of the mould can be 80 to 120°C.

[0164] In some embodiments, and with reference to FIG. 3 , the method 10 comprises adjusting 18A, 18B the mould from a first configuration used to initially mould 16 the substrate 102 or the support member 106 (e.g. initially injection mould 16) to a second configuration used to subsequently mould 12; 12A, 12B whichever of the substrate 102 and the support member 106 has not been moulded on the initially moulded part (e.g. subsequently injection mould 12; 12A, 12B whichever of the substrate 102 and the support member 106 has not been moulded on the initially moulded part).

[0165] The mold can include a first mold part and a second mold part, wherein the substrate 102 or the support member 106 can be molded between the first mold part and the second mold part. The mold can also include a third mold part, wherein whichever of the substrate 102 and the support member 106 that is not moldable between the first mold part and the second mold part is moldable between the third part and the component that has already been molded between the first mold part and the second mold part.

[0166] In such embodiments, adjusting 18A, 18B the mold from the first configuration to the second configuration can include switching the second mold part with the third mold part while the component that was molded between the first mold part and the second mold part remains housed in the first mold part. Such switching can for example include opening 18A the mold by displacing the second mold part, after which the mold is re-closed 18B using the third mold part.

[0167] Initially molding 16 (e.g., injection molding 16) the substrate 102 or the support member 106 can include heating the material above room temperature, so that the material is moldable, e.g., injectable, into the space between the first mold part and the second mold part described above.

[0168] In such embodiments, subsequently molding 12 whichever of the substrate 102 and the support member 106 that has not yet been molded on the initially molded component can be performed before the initially molded component returns to room temperature, and preferably while the initially molded component is at least 80°C. This can help to bond the substrate 102 and the support member 106 to one another. In embodiments in which the subsequent molding 12 includes curing a precursor material, such maintenance of an elevated temperature can also help such curing.

[0169] In some preferred embodiments, the substrate 102 is initially molded 16 from a thermoplastic material having a glass transition temperature of at least 140°C, such as polycarbonate, polyether ether ketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and / or polyethersulfone (PES), and the support member 106 is subsequently molded 12; 12A, 12B to the substrate 102 via curing 12B of a precursor material, e.g., a silicone precursor material.

[0170] In such embodiments, the mold can be opened 18A on one side, while the initially molded substrate 102 remains in the mold. Then, a third mold part can close the mold to assume the second mold configuration, and a precursor material, e.g., a silicone precursor material, can be injected therein.

[0171] It is noted that the mold in the second configuration can be maintained at a temperature of 80°C to 120°C, and the precursor material, e.g., a silicone precursor material, can be injected while the mold is at this temperature.

[0172] Once filled with the precursor material (e.g. a siloxane precursor material), the mold can be heated at 160 to 200 °C until the precursor material is cured, forming the support member 106, and the support member 106 is adhered to the substrate 102, e.g. the thermoplastic of the substrate 102.

[0173] More generally, when in the mold, the elastomeric material can be molded at least partially against a surface of the substrate 102, e.g. the thermoplastic substrate 102.

[0174] The mold can then be cooled to, for example, between 40 °C and 80 °C, and the two-part product can be released from the mold at this temperature. As mentioned before, this can mitigate the risk of the two-part product sticking to the mold.

[0175] The molding method disclosed herein has been described with reference to FIG. 1A to D and FIG. 2A and 2B The method is, however, not limited to the manufacture of such devices, but can be used for many other devices for many biological testing purposes. Some examples will be described below.

[0176] It is reiterated at this point that in embodiments in which several chamber units 104 are defined in the substrate 102, the chamber units 104 can be arranged in multiple rows 105. In some embodiments, for example in the embodiment shown in FIG. 4 twelve rows 105 of chamber units 104 are defined in the substrate 102, wherein each row comprises eight chamber units 104, similar to the rows of chamber units 104 shown in FIG. 1A and FIG. 1B .

[0177] The device 100 can thus have a ninety-six chamber unit 104 (e.g. well) configuration. In other embodiments, the device 100 has a twenty-four chamber unit 104 (e.g. well) configuration. The number of chamber units 104 defined in the substrate 102, as well as the shape and dimensions of the substrate 102 (e.g. and of the device 100 as a whole) can correspond to the number of wells, the dimensions and shape of a well plate as is well-known and commonly used in the art.

[0178] It is also reiterated that in some embodiments, for example in the embodiment shown in FIG. 4 a recess 114 is defined in the support member 106 for at least some (e.g. each) of the chamber units 104, and / or the support member 106 comprises at least one structural feature 112, e.g. a flexible protrusion, for contacting a biological material accommodated in at least some (e.g. each) of the chamber units 104.

[0179] FIG. 5A to FIG. 5CVarious views of a biomaterial testing apparatus 100 according to examples are provided, the biomaterial testing apparatus having a substrate 102 designed similar to the design shown in FIG. 4 However, for each chamber unit 104, the support member 106 has an end portion 110 described above with respect to the example shown in FIG. 2A The molding 12 enables the substrate 102 and the support member 106 to be attached to each other as described previously. However, for the sake of clarity, the substrate 102 and the support member 106 are shown detached from each other in the exploded perspective view provided in FIG. 5A The substrate 102 and the support member 106 are shown attached to each other in the transparent view of the apparatus 100 provided in FIG. 5B and the plan view provided in FIG. 5C

[0180] In some embodiments, for example in the embodiment shown in FIG. 6 The biomaterial testing apparatus 100 comprises a base member 132 (e.g. a glass base member 132) on which the support member 106 and the substrate 102 are arranged.

[0181] The base member 132 can help reinforce the apparatus 100. Alternatively or additionally, the base member 132 (e.g. the glass base member 132) can be optically transparent to enable analysis of the biomaterial via optical microscopy through the glass base member 132.

[0182] For some embodiments, the base member 132 (e.g. the glass base member 132) can also provide a smooth surface on which the support member 106 is molded. Such a smooth surface can in turn help make the outer surface of the support member 106 have a smooth surface. This can facilitate analysis of the biomaterial via optical microscopy through the support member 106.

[0183] It is noted that in other embodiments in which a base member 132 is not included in the apparatus 100 or, more generally, the outer surface of the support member 106 is in contact with the surface of the mold during manufacturing, the surface of the mold is preferably a polished surface.

[0184] Such a polished surface can help provide a relatively smooth outer surface of the support member 106 which can ultimately facilitate analysis of the biomaterial passing through the support member 106 via optical microscopy.

[0185] ​It is noted that at this point the focal length of many optical microscopy setups, e.g. for imaging staining, can be only about 0.6 mm. Thus, the distance from the bottom of the base member 132, e.g. the glass base member 132, and the support member 106 can not exceed 0.6 mm. However, other thicknesses can be used for different optical setups. Further details of such optical setups and options for floor materials and designs are disclosed in the currently unpublished pending patent applications EP 23181319.7 and PCT / EP2024 / 066936 as well as EP 23182831.0 and PCT / EP2024 / 067028 (all of which are incorporated herein by reference in their entirety), but other devices are possible as well.

[0186] The thickness of the optically transparent base member 132, e.g. the glass base member 132, can be 0.10 mm to 0.30 mm, e.g. about 0.20 mm.

[0187] Such thickness can take into account the thickness of the support member 106, e.g. including the height H of the flexible protrusion included in the support member 106.

[0188] More generally, the method 10 can include arranging at least a portion of the base member 132 in or adjacent to a mold, thereby initially molding 16 the substrate 102 or the support member 106 on the base member 132. After molding 12 of whichever of the substrate 102 and the support member 106 has not yet been molded on the initially molded component, the base member 132 on which the substrate 102 and the support member 106 are molded can be removed, e.g. released, from the mold.

[0189] In some embodiments, e.g. in the embodiment shown in FIG. 7 and FIG. 8 The at least one double-chamber cell 104 is defined in the substrate 102, wherein each of the double-chamber cell(s) 104 includes a first chamber 134 and a second chamber 136 fluidically connected to each other via a bridge 138 defined between the first chamber 134 and the second chamber 136, which can be referred to as a top fluidic channel. Further, the double-chamber cells 104 can be interconnected in one device by fluidic channels taking a serial, parallel or mixed configuration. For example, in FIG. 8 each row 105 of multiple double-chamber cells is interconnected in parallel between a first channel 137 and a second channel 139. The first channel can be referred to as a fluid supply channel and the second channel can be referred to as a fluid discharge channel. However, as mentioned above, other configurations can be defined. In FIG. 7 and FIG. 8In the device shown in Fig. 1, the fluidic channels 137, 138 and 139 are part of the substrate 102. They can be referred to as top channels of the device.

[0190] Such a dual-chamber unit 104 can be used to study metastasis of cancer cells. Cells released or emitted by any cell culture provided in the first chamber 134 can be evaluated. Thus, the first chamber 134 can be used to perform a treatment on the cell culture(s), e.g. to apply a certain treatment fluid or drug, e.g. a chemotherapy treatment fluid, wherein the second chamber 136 captures any released cells to analyze the effect of the treatment performed in the first chamber 134 on metastasis.

[0191] In such embodiments, the support member 106 can comprise (e.g. be defined by) a porous support member as described above for supporting biological material, e.g. cells, contained in the dual-chamber unit 104. The support member 106 may, for example, comprise a first porous portion for supporting biological material, e.g. cells, in the first chamber 134 and a second porous portion for supporting biological material, e.g. cells, in the second chamber 136.

[0192] Any suitable number of dual-chamber units 104 can be defined in the substrate 102, e.g. FIG. 7 two dual-chamber units 104 in the embodiment shown in Fig. 1 and FIG. 8 twenty-four dual-chamber units 104 in the embodiment shown in Fig. 2.

[0193] Further examples and more detailed designs of such dual-chamber unit devices have been described in the non-prepublished patent applications EP 23181319.7 and PCT / EP2024 / 066936, which are hereby incorporated by reference in their entirety.

[0194] The method 10 according to any one of the embodiments described herein can be used to form a substrate 102 comprising such dual-chamber unit(s) of a device 100 and a support member 106.

[0195] More generally, the method 10 can be used to form any device for testing biological material having at least a substrate and a support member as defined herein. Some examples of devices have been described in EP 23181319.7 and PCT / EP2024 / 066936 as well as EP 23182831.0 and PCT / EP2024 / 067028 (all of which are hereby incorporated by reference in their entirety), but other devices are possible as well.

[0196] FIG. 9A to FIG. 9FExamples are used to provide how the method 10 can be applied to manufacture a device with one or more top channels and / or one or more bottom channels, such as for example used in the device of FIG. 8 , with top channels 137, 138 and 139 and other features in the device. FIG. 9A To F are also used to illustrate a mold system with different parts to be used in the method. Although FIG. 9A to 9G Examples are shown with a substrate and support member having two chamber units, the figures can be considered to also represent cases where more of such chamber units are present, for example in the devices of FIG. 5A to 5C , 6 and 8.

[0197] FIG. 9A to FIG. 9F A method is illustrated to manufacture a three-part multi-chamber device 900 with chamber units connected in parallel, which device is very similar to the device of FIG. 8 , with the difference that the device 900 will have only one chamber (or vessel) for each chamber unit, compared to the chamber units of FIG. 7 and FIG. 8 .

[0198] FIG. 9E A cross-sectional view of the device 900 to be manufactured is shown, and FIG. 9F and FIG. 9G corresponding bottom and top views of the device 900 are shown. FIG. 9E Three cross-sectional views of the device 900 are shown. The two lower views IX and X show cross-sections perpendicular to the cross-section of the top view, along the directions IX and X indicated therein.

[0199] The device 900 comprises two chamber units 934 within a substrate 902, each having a cylindrical shape with a diameter 170. Other shapes can be used. The circumference of the chamber side wall is indicated in dashed format in FIG. 9F , as the chambers cannot be seen from the bottom side of the device 900 through the support member 906 in this view.

[0200] The device further comprises two top channels 937 and 939, which can be seen in the top view of FIG. 9G to extend horizontally in the substrate 902. The chamber units 934 are connected to the channels 937 and 939 via channels 938, such that the chambers are in a parallel configuration.

[0201] On the bottom side of the device 900, there is a support member 902 adhered to the substrate 902 according to the principles as disclosed herein.

[0202] The support member 906 comprises a bottom channel 935 connecting sub-chambers 975 located below a membrane 942. In FIG. 9FIn a top view, the channels 935 can be seen extending horizontally.

[0203] The support member 902 comprises end portions 910, each of which is located within one of the chamber units 934. Each end portion comprises a membrane 942 having a circular shape with a diameter 972 and a membrane thickness measured perpendicular to the plane in which it extends. In FIG. 9F and 9G The circular shape can be seen from above or below. Each end portion further comprises a flange 966, each of which extends upwards from the bottom side of the base plate 902 into the chamber unit 934, having a certain height which is smaller than the height of the chamber unit 934. Each of the membranes 942 extends from, or in other words is carried by or integrated with, the flange 966.

[0204] The preferred membrane thickness is in the range of 5 pm to 100 pm, but other thicknesses can be used. This can be chosen based on the desired purpose of the device.

[0205] In some embodiments, the membrane comprises or includes a perforated membrane comprising a plurality of pores. Each pore is preferably wide enough to allow passage of a fluid which can contain nutrients, drugs or other (bio)molecules, such as proteins and the like.

[0206] In some applications, the pores are wide enough to at least partially allow passage of cells or a specific type of cells.

[0207] In some embodiments, the pores are narrow enough to prevent passage of cells. For example, the pore diameter can be in the range of 10 pm or less, for example it can be 5 pm. But other values as described below can be used. Thus, the porous membrane can be used to support a tissue sample, such as cells, while a fluid having all of its content as indicated above can be provided to the cells by passing through the membrane using one or more of the channels in the device. Exemplary membranes and channels are in the device of FIG. 7 and FIG. 8 and the device described in EP23181319.7 and PCT / EP2024 / 066936, for example.

[0208] The support member 902 can have other structural features 912 in its end portions 934. Thus, for example, a plurality of holes can be defined in the membrane. The bottom of such holes can again form a membrane or perforated membrane as described herein. Detailed examples of such membranes comprising holes have been described in EP23181319.7 and PCT / EP2024 / 066936.

[0209] The pores can be made, for example, using laser perforation as disclosed herein.

[0210] FIG. 9A to 9DEach shows three cross-sectional views of a mold system as defined herein for use in a method as defined herein to manufacture a device 900. It will be appreciated that the mold system can be used to manufacture other devices by adapting its structural design. The mold system comprises a first mold part 950 and a second mold part 952, which can be combined to form a first configuration for the initial molding step 16 as previously described herein. The first and second mold parts have a shape and size designed to mold the substrate 902 of the device 900. The two following views I and II show cross-sections perpendicular to the top view along the direction indicated by I and II.

[0211] The first and second mold parts together are geometrically designed and shaped to have protrusions and open spaces, sometimes referred to as one or more first open spaces, which together define the shape and design of the substrate 902 during and after molding. In this case, the first and second mold parts have protrusions 953 which at least partly define the chamber cells 934 and protrusions 955 which ultimately define the top channel in the substrate 902, very similar to the channels 137 and 139 of the device of FIG. 8 . The top channel will be defined by filling the open spaces 954. In more detail, the protrusions 953 are in this case cylindrical, have a diameter 170, and their cylinder axes are vertical and in the plane of the cross-sectional view. After molding, they will ultimately define cylindrical chamber cells 934 in the substrate 902 having a diameter 170. The protrusions 955 are rectangular and extend along the first mold perpendicular to the plane of the view I of the cross-sectional view. The first mold part 950 shows protrusions 957 abutting the protrusions 953. The protrusions 957 are rectangular and extend only partly along the cylindrical protrusions 953, so that ultimately they will form the top channel connecting the chambers 934 to the top channel. The second mold part 952 has surface mold surface portions 980, sometimes referred to as second mold surfaces, which will define substrate surfaces 982 which at a later stage of the process will define the part of the substrate onto which the support member 906 will be molded. These surfaces 982 will thus be the contact or adhesive surfaces as previously described. The mold surface portions 980 can be geometrically designed to improve or increase the adhesion of the support member to the substrate. In FIG. 9A to FIG. 9G , for the sake of clarity, no specific geometric design is shown, but the first mold part surface 980 can have for example a specific surface roughness and / or some kind of corrugation.

[0212] The first and second mold parts in FIG. 9AThe middle is shown as being separated from each other in an open configuration, and they are releasably attachable to each other in a leak-proof manner (separable from each other), such that in the first mold configuration (when they are combined and held together), as FIG. 9B and FIG. 9D shown, they can be used to mold by injecting a liquid material therein. For example, they can be clamped together to form a closed first mold configuration. The first mold part has an injection opening 156 for inserting a liquid substrate material into the mold when the mold is in the closed molding configuration.

[0213] In the initial molding step 16 as described with reference to method 10 and FIG. 3 the first mold part 950 and the second mold part 952 are brought into the first configuration, and under the described conditions, the substrate material is molded into the substrate 902 by injection molding the material into the mold opening 954, for example, via the injection opening 156. The result is shown in cross-sectional view in FIG. 9B As an example, the injected material can be a material selected from the group of thermoplastic materials, for example, polycarbonate, as described earlier herein.

[0214] In a further step 18A, the second mold part 952 is released from the first mold part 950 and the molded substrate 902, while the substrate 902 is still within the first mold part 950. The result is shown in FIG. 9C The design of the protrusions 953 of the first and second mold parts 952, 953 now results in the overall first mold part and substrate 902 having open spaces 964, which open spaces have a diameter 170 and each form an exposed portion 964 of the substrate chamber 134. These open spaces will eventually be occupied by the end portion 910 of the support material once molded. In addition, the substrate.

[0215] In step 18B, the second mold part 952 is replaced with a third mold part 962, which is designed to fit to the first mold part 950 to form a second mold configuration in which additional open spaces (sometimes referred to as one or more second open spaces) are retained in which the support member 906 is to be molded. The design of the first mold part 950, the second mold part 960, and the molded substrate 902 now defines additional open spaces. This second mold configuration is shown in FIG. 9D

[0216] ​The third mold part 962 comprises protrusions 961 having a cylindrical shape with a diameter 972 and a cylindrical axis that is oriented vertically and in the plane of the drawing. The diameter 972 is smaller than the diameter 970 of the protrusions 953 of the first mold part 950, and the height of the protrusions is such that they do not contact the protrusions 953 of the mold part 950 in the second closed configuration. An open space in the form of an exposed portion 964 is thus left in each of the chamber cells 934, which exposed portion can be occupied by the material that will form the end portion of the substrate member 906. In this case, the additional open space is thus designed to result in a support member 906 with a membrane 942 attached to the flange 966, with the membrane being between the protrusions 953 and 961. However, other designs can be used, and the design of the protrusions 953 and 961 can be used to define a desired end portion, such as those defined herein. In this context, the surface 964 can be referred to as a first mold surface, and the surface 986 of the protrusions 961 can be referred to as a third mold surface.

[0217] The third mold part also has one or more protrusions (only one shown for clarity) 963 having a rectangular shape and extending perpendicular to the drawing of the cross-sectional view VII. These protrusions will eventually form channels 935 in the support member 906 that interconnect the sub-chambers 975 under the membrane 942.

[0218] In step 12, when the second mold configuration has been assumed, the support member 906 can be molded as described herein with reference to method 10 and FIG. 3 The material for molding is injected through the openings 960 to form the result as shown in FIG. 9D When the support member material is molded against the portion of the surface of the substrate, the contact between the contacting surfaces is intimate, and the support member is molded to be highly conformal to the shape and irregularities in the surface of the substrate 902. This results in intimate contact with improved adhesion and leakage properties, as described above.

[0219] After cooling to an appropriate temperature as indicated herein before for method 10, the device in the form of the support member 906 molded onto the substrate 902 can be released from the first and second mold parts. The result will be a device 900 as described above FIG. 9E to FIG. 9F If no further parts are needed, the device can be used as such. This can be the case, for example, if the support member has a design without any bottom channels, such as channels 935 or other openings that need to contain a fluid. A device with posts as described herein can provide such an example. However, FIG. 9E The device of does have bottom channels, and these can need to be closed from the bottom side.

[0220] ReferenceFIG. 9A to FIG. 9D The mold system used by the method described in the method of the first aspect comprises a first mold part, a second mold part and a third mold part as defined herein.

[0221] In an alternative method according to the application, the molding of the substrate and the molding of the support member are not combined. Thus, the substrate is an already existing substrate, which can have been molded in a stationary manner, but is provided from stock. The first mold part is then designed as defined herein, and accommodates the substrate provided from stock. The first mold part and the second mold part are designed and used as referred to FIG. 9A to FIG. 9D In such alternative method, the mold system according to the present disclosure then only comprises a first mold part and a third mold part as defined herein.

[0222] To close the bottom channel, the device can be adhered to a base member 932 made of, for example, glass or other suitable material, such as those described herein. FIG. 9H The device of FIG. 9E to FIG. 9G is shown, which has a base member 932 that closes the channel 935 and the opening 975 on the bottom side of the device, so that they are suitable to contain fluid. The channel is now suitable to provide fluid to the membrane 942 from the bottom side, or to provide the exhaust fluid that has passed through such membrane from the top to the bottom side.

[0223] In an alternative method, the base member is already present in the third mold part 962, for example as an inlay or pre-molded in the third mold part 962, before the mold material of the substrate member 906 is injected. In this case, the support member can not only be molded to be conformal with the substrate 902, but can also be molded to be conformal with the base member 932.

[0224] A top cover plate can be added as desired. This can be releasably attachable to the substrate 902. The top cover plate can be glass or a polymeric material. It can be the same material as the material of the substrate. For the sake of clarity, the top cover is not shown in FIG. 9H .

[0225] It will be clear that a device of the two- or multi-component type as described herein can have a desired design that differs from the examples provided herein. Thus, by adjusting the design and dimensions of the protrusions 935 and 961, the size and shape of the chamber units 934 can be defined. Furthermore, their design and dimensions can also be used to define the open space between the protrusions 935 and 961 to thereby define the shape and dimensions of the end portion 91o and the membrane 942. For example, the thickness of the membrane can be set with these design parameters. The membrane can also have a hole defined therein. More detailed designs of membranes with perforations and / or holes have been described in reference to other documents provided herein (including their desired dimensions). Also, the device 900 has only one chamber for each chamber unit. As for the device 800, FIG. 8 Two or more chambers can be present for each chamber unit. Fig. 9 shows a device 900 with only two chambers and corresponding channels. However, this is only exemplary, as the principles shown can be extended to the manufacture of devices with different numbers of chambers and different numbers of channels, etc. Thus, for example, a 96-chamber plate can be manufactured with this approach.

[0226] More generally, the present disclosure proposes to test a biological material, for example a test cell group, a bilayer of cells, spheroids, organoids or a biopsy, using a device 100 according to any of the embodiments described herein.

[0227] The present disclosure also contemplates a drug testing method comprising providing a biological material in at least one chamber unit 104 of a device 100 according to any of the embodiments described herein, and exposing the biological material to a drug to be tested.

[0228] In some embodiments, providing a biological material in at least one chamber unit 104 comprises culturing cells in at least one chamber unit 104.

[0229] Alternatively or additionally, the drug testing method can comprise monitoring the response of the biological material, for example the cultured cells, to the drug to be tested. This monitoring can be implemented in any suitable way, for example by optical microscopy, for example through the support member 106 when the support member 106 is optically transparent.

[0230] A preferred combination of silicone and (rigid) thermoplastic is silicone / polycarbonate, wherein the glass transition temperature (Tg) of the polycarbonate is > 140 °C or silicone / PEEK and silicone / PEEK blends:

[0231] In certain embodiments, the following exemplary commercially available elastomer / soft material (which is transparent or translucent and is food contact approved) can be used as a material for the support member 106: Medalist MD-53253 (TPE Teknor Apex); Medalist MD-53273 (TPE Teknor Apex); Mediprene 500602 M-03 (TPE Hexpol); Texin Rx T85A (TPU Covestro); BioSpan® (SPU | PUR); BioSpan® (SPU | PUR); BJB Polyurethane F-116 A / B | TSU; BJB Polyurethane F-126 A / B | TSU; BJB Polyurethane F-131 A / B | TSU; BJB Polyurethane M-3115 REV 1 A / B | TSU; BJB Polyurethane M-3125 A / B | TSU; CELLENE MC2248 | TPE; CELLENE MC2265 | TPE; CELLENE MC3038 | TPE; CELLENE MC3050 | TPE; CELLENE MC3061| TPE; CELLENE MC3226 | TPE; CELLENE MC3239 | TPE; CELLENE MC3261 | TPE; Dynaflex™ G2706-1000-00 | TPE; Dynaflex™ G2711-1000-00 | TPE; Elastocon® 2860L | TPE; Filter-bond™ E-3264 | TS; FLEXCHEM™ 3551-02 | PVC, Flexible; FLEXCHEM™ 4051-02 | PVC, Flexible; FLEXCHEM™ 4551-02 | PVC, Flexible; FLEXCHEM™ 5051-02 | PVC, Flexible; FLEXCHEM™ 5551-02 | PVC, Flexible; FLEXCHEM™ 6051-02 | PVC, Flexible; FLEXCHEM™ 6551-02 | PVC, Flexible; Medalist® MD-12130 | TPE; Medalist® MD-12130H | TPE; Medalist® MD-12140 | TPE; Medalist® MD-12140H | TPE; Medalist® MD-12150 | TPE; Medalist® MD-12150H | TPE; Medalist® MD-12f50S | TPE;Medalist® MD-12160 | TPE; Medalist® MD-12160H | TPE; Medalist® MD-12170 | TPE; Medalist® MD-12170H | TPE; Medalist® MD-12243 | TPE; Medalist® MD-12337 | TPE; Medalist® MD-12340NAT | TPE; Medalist® MD-12342 | TPE; Medalist® MD-12344 | TPE; Medalist® MD-12350 | TPE; Medalist® MD-12352 | TPE; Medalist® MD-12362 | TPE; Medalist® MD-125 | TPE; Medalist® MD-130 | TPE; Medalist® MD-13240 | TPE; Medalist® MD-135 | TPE; Medalist® MD-145 | TPE; Medalist® MD-155 | TPE; Medalist® MD-17365 | TPE; Medalist® MD-225 | TPV; Medalist® MD-32045 | TPE; Medalist® MD-32245 | TPE; Medalist® MD-36048 | TPE; Medalist® MD-37063 NAT | TPE; Medalist® MD-42245 XRD1 | TPE; Medalist® MD-42245 XRD3 | TPE; Medalist® MD-74357 XRD1 | TPE; Medalist® MD-74357 XRD2 | TPE; Mediprene® 500120M | TPE; Mediprene® 500200M | TPE; Mediprene® 500250M | TPE; Mediprene® 500300M | TPE; Mediprene® 500350M | TPE; Mediprene® 500400M | TPE; Mediprene® 500434M | TPE; Mediprene® 500450M | TPE; Mediprene® 500484M | TPE; Mediprene® 500520M | TPE; Mediprene® 500534M | TPE;Mediprene® 500584M | TPE; Mediprene® 500600M | TPE; Mediprene® 500634M | TPE; Mediprene® 500650M | TPE; Mediprene® 500684M | TPE; Mediprene® 500700M | TPE; Monprene® RG-10160H | TPE; ProvaMed® TPE 1120 | TPE; ProvaMed® TPE 1160 | TPE; RABALON® PJ4300C | TPE; RABALON® PJ5300C | TPE; RABALON® PJ6300C | TPE; RABALON® PJ7300C | TPE; SkinFlex 15 F-115 A / B | TSU; SkinFlex BR-60; BRUSHABLE A / B | TSU; T-Blend® TPE-F22 | SEBS; THERMOLAST® M TM3LFT (series: MC / LF) | TPE; THERMOLAST® M TM3MED (series: MC / tl) | TPE; THERMOLAST® M TM3RST (series: MC / RS) | TPE; THERMOLAST® M TM4LFT (series: MC / LF) | TPE; THERMOLAST® M TM4MED (series: MC / tl) | TPE; THERMOLAST® M TM4RST (series: MC / RS) | TPE; THERMOLAST® M TM5LFT (series: MC / LF) | TPE; THERMOLAST® M TM5MED (series: MC / tl) | TPE; THERMOLAST® M TM6LFT (series: MC / LF) | TPE; THERMOLAST® M TM6MED (series: MC / tl) | TPE; THERMOLAST® M TM7LFT (series: MC / LF) | TPE; THERMOLAST® M TM7MED (series: MC / tl) | TPE; UNISOFT SPECIAL™ DS-35A-CL-M-01 | SEBS; UNISOFT SPECIAL™ DS-55A-CL-M-01 | SEBS; Versaflex™ G2705 N | TPE; Versaflex™ HC 1100-40 Translucent EU | TPE;Versaflex™ HC 1348 Natural | TPE; Versaflex™ HC MT317 | TPE; Versaflex™ HC MT555 | TPE; Versaflex™ OM 1040X-1 | TPE; CELLENE MC2248 | TPE; CELLENE MC2265 | TPE; CELLENE MC3038 | TPE; CELLENE MC3050 | TPE; CELLENE MC3061 | TPE; CELLENE MC3226 | TPE; CELLENE MC3239 | TPE; CELLENE MC3261 | TPE; ChronoPrene™ 25A | TPE; ChronoPrene™ 40A | TPE (CardioTech International, Inc.); Dryflex® 500300S | TPE; Dryflex® 500350S | TPE; Dryflex® 500400S | TPE; Dryflex® 500450S | TPE; Dryflex® 500500S | TPE; Dryflex® 500550S | TPE; Dryflex® 500600S | TPE; Dryflex® 500650S | TPE; Dryflex® 500700S | TPE; Dynaflex™ G2701-1000-02 | TPE; Dynaflex™ G2706-1000-00 | TPE; Dynaflex™ G2709-1000-00 | TPE; Dynaflex™ G2711-1000-00 | TPE; Dynaflex™ G2712-1000-02 | TPE; Dynaflex™ G2730 | TPE; Dynaflex™ G2755-1000-00 | TPE; Dynaflex™ G2755C | TPE; Dynaflex™ G6713-0001 | TPE; Dynaflex™ G6713C | TPE; Dynalloy™ GP 7810-60T | TPE; Dynalloy™ GP 7810-70T | TPE; Dynalloy™ OBC8200-BT50 | TPE; Estane® 58123 TPU | TPU-Polyether; Evoprene™ 019 | SBS; Evoprene™ G 925 | SEBS;Evoprene™ G 936 | SEBS; Evoprene™ G 942 | SEBS; Evoprene™ G 958 | SEBS; Evoprene™ G 966 | SEBS; Evoprene™ G 967 | SEBS; Evoprene™ G 968 | SEBS; Evoprene™ G 969 | SEBS; Evoprene™ G 970 | SEBS; Evoprene™ GC 5685 | SEBS; Evoprene™ GC 5686 | SEBS; Evoprene™ GC 5687 | SEBS; Evoprene™ GC 5688 | SEBS; Evoprene™ GC 5689 | SEBS; Evoprene™ GC 5690 | SEBS; GLS 422-126 | TPE; GLS 458-140 | TPE; GLS 458-141 | TPE; GLS 458-142 | TPE; K-Prene HYFLEX HF 15 | MPR; K-Prene HYFLEX HF 20 | MPR; K-Prene HYFLEX HF 25 | MPR; K-Prene HYFLEX HF 30 | MPR; Medalist® RG-38052XRD1 | TPE; megol® PUG 10 | SEBS; megol® PUG 60 | SEBS; megol® TA 60 | SEBS; Monprene® RG-10130 | TPE; Monprene® RG-10140 | TPE; Monprene® RG-10150 | TPE; Monprene® RG-10160 | TPE; Monprene® RG-10170 | TPE; Monprene® RG-15130 | TPE; Monprene® RG-15140 | TPE; Monprene® RG-15150 | TPE; Monprene® RG-15160 | TPE; Monprene® RG-15170 | TPE; Monprene® RG-18240 | TPE; Monprene® RG-18250 | TPE; Monprene® RG-18260 | TPE; Monprene® RG-18270 | TPE; Monprene® RG-19221NAT | TPE;Monprene® RG-19255 | TPE; Monprene® RG-20140 | TPE; Monprene® RG-20160 | TPE; Monprene® RG-20170 | TPE; Monprene® RG-29068 NAT | TPE; Monprene® RG-29240 XRD1 | TPE; RABALON® MJ4300C | TPE; RABALON® MJ5302C | TPE; RABALON® MJ6301C | TPE; RABALON® MJ7301C | TPE; RAYPRENE® NB221-S4050 | TPE; RAYPRENE® NB221-S4051 | TPE; RAYPRENE® NB221-S4052 | TPE; RAYPRENE® NB221-S4053 | TPE; tefabloc® TO 132 | TPE; Telcar® TL-83-F943D22-NT BLU | TPE; THERMOLAST® K TF2CGT (series: FC) | TPE; THERMOLAST® K TF3BTL (series: FC / AP) | TPE; THERMOLAST® K TF3CGT (series: FC) | TPE; THERMOLAST® K TF3STE (series: FC / CS) | TPE; THERMOLAST® K TF4AAB (series: FC / HE / tl) | TPE; THERMOLAST® K TF4BTL (series: FC / AP) | TPE; THERMOLAST® K TF4CGT (series: FC) | TPE; THERMOLAST® K TF4STE (series: FC / CS) | TPE; THERMOLAST® K TF5AAC (series: FC / HE / tl) | TPE; THERMOLAST® K TF5BTL (series: FC / AP) | TPE; THERMOLAST® K TF5CGT (series: FC) | TPE; THERMOLAST® K TF5STE (series: FC / CS) | TPE; THERMOLAST® K TF5WHA (series: DW / H) | TPE; THERMOLAST® K TF6AAF (series: FC / HE / tl) | TPE; THERMOLAST® K TF6BTL (series: FC / AP) | TPE;K TF6CGT (series: FC) | TPE; THERMOLAST® K TF6STE (series: FC / CS) | TPE; THERMOLAST® K TF6WCS (series: DW / CS) | TPE; THERMOLAST® K TF6WHA (series: DW / H) | TPE; THERMOLAST® K TF6WHB (series: DW / H) | TPE; THERMOLAST® K TF7AAC (series: FC / HE / tl) | TPE; THERMOLAST® K TF7BTL (series: FC / AP) | TPE; THERMOLAST® K TF7CGT (series: FC) | TPE; THERMOLAST® K TF7WHB (series: DW / H) | TPE; Topolymer® 8201-B | TPE; Versaflex™ FFC 2882-50 EU | TPE; Versaflex™ FFC 2882-50 | TPE; Versaflex™ G2708 N | TPE; Versaflex™ GP 2810-20N | TPE; Versaflex™ GP 2810-30N | TPE; Versaflex™ GP 2810-40N | TPE; Versaflex™ GP 2810-50N | TPE; Versaflex™ GP 2810-60N | TPE; Versaflex™ GP 2810-70N | TPE; Cawiton® MT920 | SEBS; Cawiton® MT930 | SEBS; Cawiton® MT940 | SEBS; Cawiton® MT950 | SEBS; Cawiton® MT960 | SEBS; Cawiton® MT970 | SEBS.

[0232] Variations to the disclosed embodiments can become apparent to those of ordinary skill in the art upon reading the foregoing description, which is provided by way of example only. In the claims, the word “comprising” does not exclude other elements or steps, and the words “a” or “an” do not exclude a plurality.

[0233] Although specific measures are recited in mutually different dependent claims, this does not indicate that a combination of these measures cannot be used to advantage.

[0234] If the term“adapted to” is used in the claims or specification it should be noted that the term“adapted to” is intended to be equivalent to the term“configured to”.

[0235] Any reference signs in the claims should not be construed as limiting the scope.

Claims

1. A method (10) for manufacturing a biomaterial testing device (100), the device comprising: a substrate (102) in which at least one chamber unit (104) is defined; and a support member (106) for supporting a biological material accommodated in the at least one chamber unit, the method comprising: moulding (12) one of the substrate and the support member on at least part of the other of the substrate and the support member using a mould, the at least part of the other of the substrate and the support member being arranged in or adjacent to the mould during the moulding.

2. The method (10) of claim 1, comprising: The substrate (102) or the support member (106) is initially moulded (16) to form an initially moulded part, and subsequently whichever of the substrate and the support member has not been moulded on the initially moulded part is moulded.

3. The method (10) according to claim 2, wherein The substrate (102) or the support member (106) is initially moulded (16) comprises heating a material above room temperature to enable the material to be moulded, thereby enabling the initially moulded part to be formed, and wherein subsequently moulding whichever of the substrate and the support member has not been moulded on the initially moulded part is performed before the initially moulded part returns to room temperature, and preferably is performed when the temperature of the initially moulded part is at least 80 °C.

4. The method (10) according to claim 2 or 3, comprising: Arranging at least part of a base member (132) in or adjacent to the mould on which the substrate (102) or the support member (106) is initially moulded, and removing the base member from the mould, the base member having the substrate and the support member moulded thereon.

5. The method (10) according to any one of claims 1 to 4, wherein Moulding (12) the one of the substrate (102) and the support member (106) on the at least part of the other of the substrate and the support member comprises curing a precursor material, optionally wherein the curing comprises heating the precursor material.

6. The method (10) according to any one of claims 1 to 5, comprising: Releasing the substrate (102) from the mould together with the support member (106) when the temperature of the substrate and the support member is above room temperature and preferably is at least 40 °C.

7. The method (10) according to any one of claims 1 to 6, comprising: Injection moulding (12) the one of the substrate (102) and the support member (106) on the at least part of the other of the substrate and the support member.

8. The method (10) according to any one of claims 1 to 7, comprising: Adjusting (18A, 18B) the mould from a first configuration, which is used to mould the one of the substrate (102) and the support member (106), to a second configuration, which is used to mould the other of the substrate and the support member.

9. The method (10) according to any one of claims 1 to 8, wherein The support member (106) is formed from a material that is softer and / or more flexible than the material from which the substrate (102) is formed.

10. The method (10) according to any one of claims 1 to 9, wherein The support member (106) is formed from a material that is more biocompatible than the material from which the substrate (102) is formed, such that cells and / or tissue preferably adhere to the support member.

11. The method (10) according to any one of claims 1 to 10, wherein The support member (106) comprises silicone.

12. The method (10) according to any one of claims 1 to 11, wherein The support member (106) comprises a polymeric material bulk-modified with a moiety comprising a polar group, the polar group of the moiety being obtainable at a surface of the support member, the surface being arranged to contact the biological material accommodated in the at least one chamber cell (104); optionally wherein the moiety comprises a fatty acid moiety, the polar group comprising a carboxylic acid group of the fatty acid moiety.

13. The method (10) according to any one of claims 1 to 12, wherein The substrate (102) is formed of a material having a glass transition temperature of at least 140 °C; and / or wherein the substrate is formed of a thermoplastic, preferably a thermoplastic selected from one or more of the following: polycarbonate, polyether ether ketone, acrylonitrile butadiene styrene, polyetherimide, and polyethersulfone.

14. The method (10) according to any one of claims 1 to 13, wherein The substrate (102) provides a side wall(s) (108) of the at least one chamber cell (104), a central portion (124) of the support member is provided in the at least one chamber cell, and a plurality of rib elements (126) each extend from the central portion to engage with the side wall of the respective chamber cell.

15. The method (10) according to any one of claims 1 to 14, wherein The support member (106) comprises at least one structural feature (112) for contacting the biological material accommodated in the at least one chamber cell (104); optionally wherein the at least one structural feature comprises, for each chamber cell, a pair of flexible protrusions for supporting tissue thereon and therebetween.

16. The method (10) according to any one of claims 1 to 15, wherein The at least one chamber cell (104) comprises a plurality of chamber cells; optionally wherein the chamber cells are arranged in one or more rows (105).

17. A mold system for molding a biological testing device using the methods disclosed herein, the mold system comprising: a first mould part; and a third mould part, the third mould part being separable from the first mould part; wherein the first mould part and the third mould part are configured to be combined to form a second mould configuration, the second mould configuration being arranged to hold one of a substrate and a support member, such that in the second mould configuration the first mould part, the second mould part, and the one of the substrate and the support member define one or more openings in which the other of the substrate and the support member can be moulded onto the one of the substrate and the support member.

18. A biological material testing apparatus (100) comprising: a substrate (102) in which at least one chamber cell (104) is defined; and a support member (106) for supporting a biological material accommodated in the at least one chamber cell, one of the substrate and the support member being moulded onto the other of the substrate and the support member.

19. Use of the apparatus (100) of claim 18 for testing a biological material.

20. A method of testing a drug comprising: providing a biological material in at least one chamber unit (104) of the apparatus (100) according to claim 18, optionally wherein said providing a biological material in said at least one chamber unit comprises culturing cells in said at least one chamber unit; and exposing said biological material to a drug to be tested.

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