Biological material testing device and methods for manufacturing and using the same

Abstract

Provided is a biological material testing device (100) comprising a substrate (102) in which at least one chamber unit (104) is defined, and a support member (106) for supporting biological material received 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. Also provided is a method (10) for manufacturing such a biological material testing device. Further provided are uses and methods in which the device is employed for testing biological material, for example in drug testing in which the biological material is exposed to a drug to be tested.

Description

[0001] BIOLOGICAL MATERIAL TESTING DEVICE AND METHODS FOR MANUFACTURING AND

[0002] USING THE SAME

[0003] FIELD OF THE INVENTION

[0004] The invention relates to a method for manufacturing a biological material testing device.

[0005] The invention also relates to the biological material testing device per se and the biological testing device obtainable by the method.

[0006] The invention also relates to a mold for use in the manufacturing method.

[0007] The invention further relates to use of such a device for testing biological material.

[0008] The invention yet further relates to a drug testing method in which the device is used.

[0009] BACKGROUND OF THE INVENTION

[0010] In vitro testing of mammalian cells or tissue is an important technique to obtain clinically important information of the mammalian material under investigation. For example, biopsied mammalian cell or tissue material may be subjected to such testing to determine anomalies or disease in such mammalian material or to expose diseased mammalian material to drugs, e.g., experimental drugs, to monitor the response of the diseased mammalian material to such exposure. This approach, for example, is frequently used in oncological procedures. This can provide important insights in how a 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 number of reasons, including drug toxicity.

[0011] Drugs may be tested for cardiotoxicity. Many drugs can deleteriously affect heart muscle contraction and profile. Drug-induced cardiotoxicity is a major adverse effect that has been encountered for some clinically important drugs. This toxicity has previously led to post-marketing withdrawal of numerous pharmacologically active drugs and has limited the efficacy of other clinically useful ones. Almost 10% of drugs in the last four decades have been recalled from the market worldwide due to cardiovascular safety concerns (see, for example, “Drug Induced Cardiotoxicity: Mechanism, Prevention and Management” by Mina T. Kelleni and Mahrous Abdelbasset, 2018; a chapter of “Cardiotoxicity”, edited by Wenyong Tan).

[0012] Hence drug-induced cardiotoxicity represents a key reason for rejecting compounds in preclinical and clinical development, which reflects the seriousness of cardiotoxicity as a side effect observed during novel drug development. Assessing drug-induced cardiotoxicity risk, including QT interval prolongation, is considered nowadays an integral part of standard preclinical evaluation of new chemical entities (referring again to “Drug Induced Cardiotoxicity: Mechanism, Prevention and Management”). Cardiotoxicity may be tested in animal models, for example in rats. However, these models have drawbacks and do not always reliably predict effect(s) experienced by humans. This may result in potential beneficial medications being discarded, but may also mean that potentially toxic medications progress to human trials.

[0013] Another way to assess cardiotoxicity is to test the effect of the drug in vitro. The drug to be tested may be added to an assembly of cardiomyocytes, which are grown in vitro. Afterwards, the effect of electrical stimuli on the assembly of cardiomyocytes may be studied. An example of a state-of- the-art tool, is the “HeartDyno” (Trademark), developed by the group of James Hudson (see, 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).

[0014] This type of tool, which is encompassed herein by the term “support member”, may include an oval shaped well, with two small protrusions, e.g. pillars, at the bottom. Such a support member can be produced using thin film technologies. A mold can be made via SU-8 photolithography on a wafer, e.g., resulting in 700 pm deep features. Polydimethylsiloxane (PDMS) can then be cast over these features and cured. After removal of the PDMS from the wafer, samples of 6 mm diameter are punched.

[0015] These samples are for example placed inside the wells of a ninety six-well substrate and glued to the bottom of the substrate using silicone glue / adhesion. A mixture of cardiomyocyte cells, cardio fibroblast cells, collagen, DMEM, NaOH and matrigel can then be added to the wells and supported on samples. It is noted that the well or wells defined in such a substrate are referred to herein using the more general term “chamber unit”. The cardiomyocyte tissue is formed over several days. The tissue shows spontaneous contraction, but also contracts during electrical stimulation. During contraction, the two pillars deflect and the deflections are analyzed using video analysis algorithms.

[0016] Fluidic devices are also known that include two or more fluid channels for carrying fluids to one or more chamber units, e.g., culturing chamber(s), defined in the fluidic device’s substrate. One channel can be used, for example, to provide nutrients and oxygen to each of the chamber unit(s), as well as to remove metabolic products such as carbon dioxide therefrom. A further channel can be used to provide medication to the chamber unit(s). Such devices may include a support member, for example a porous support member, for supporting biological material, such as cell groups, double cell layers, spheroids, organoids or biopsies, received in the chamber unit(s).

[0017] SUMMARY OF THE INVENTION

[0018] Attaching the substrate, e.g., of the type mentioned above, and the support member, e.g. of the type mentioned above, to each other may present various technical challenges. It may be difficult to implement such attachment in a sterile manner and / or without inadvertently providing points between the substrate and the support member where fluid leakage can occur. Moreover, additional manufacturing steps may be necessary to attach the substrate and the support member to each other. The invention is defined by the claims.

[0019] According to examples in accordance with an aspect of the disclosure, there is provided a method for manufacturing a biological material testing device, the device comprising a substrate in which at least one chamber unit is defined, and a support member for supporting biological material received in the at least one chamber unit, the method comprising molding, using a mold, one of the substrate and the support member on at least part of the other of the substrate and the support member, which at least part of the other of the substrate and the support member is arranged in or adjacent to the mold during said molding.

[0020] By molding one of the substrate and the support member on at least part of the other of the substrate and the support member, the former may be shaped and adhered onto the latter in the same 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 may be obviated. No adhesive / glue may be required to keep the substrate and the support member attached to each other. Thus, the method may provide a relatively quick and reliable process for manufacturing the device, in which the substrate and the support member can be regarded as being integrated with each other in a single piece.

[0021] Moreover, the support member, e.g., silicone support member, may be anchored by geometry into a frame, e.g., polycarbonate frame, defined by the substrate. For example, surfaces of the substrate and of the support member that will be in contact in a device may have geometrical structures or shapes to promote adherence. Such structures or shapes may be chosen from a group consisting of: rough surfaces, corrugations, protrusions, recesses. However other structure may be used in addition or alternatively.

[0022] In at least some embodiments, the molding comprises injection molding said one of the substrate and the support member on said at least part of the other of the substrate and the support member. Injection molding may provide a cost-effective and scalable way of manufacturing the device.

[0023] In some embodiments, the method comprises initially molding the substrate or the support member to form an initially molded component, and subsequently molding whichever of the substrate and the support member has not already been molded on the initially molded component. In such embodiments the method may comprise two-step injection molding, e.g., so-called “2K” injection molding.

[0024] Alternatively, or additionally, molding of said one of the substrate and the support member on the at least part of the other of the substrate and the support member may comprise curing a precursor material. The precursor material is preferably in liquid state during its addition to the mold. In the case of injection molding, the molding may 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 cause the precursor material to transform into a solid support member material. Precursor materials that undergo chemical transformation are preferred. The use of a precursor material is advantageous for molding of a substrate and / or support member comprising a rubber material. Such materials are difficult to reshape after their formation. For example, support members as disclosed herein comprising silicone polymers of the rubber type are preferably made using a liquid precursor material that solidifies into the silicone polymer rubber through chemical reactions of the precursors components. Other rubber materials may be used too and may benefit from the use of precursor materials in the method.

[0025] The curing may comprise heating the precursor material, for example above 160°C, e.g., between 160 and 200°C. Heating at such temperatures may assist to ensure that the precursor material is cured and adhered to the at least part of the other of the substrate and the support member, e.g., the initially molded component. Heating, may be used to cause the chemical and / or physical transformations to occur.

[0026] In some embodiments, initially molding the substrate or the support member comprises heating a material above room temperature to make the material moldable to enable forming of the initially molded component, for example to 200 to 300°C in the case of a thermoplastic material, often referred to as a thermoplastic. It is noted that the term “thermoplastic material” as used herein is intended to refer to a polymer that can be softened through heating and then processed, in particular molded.

[0027] In such embodiments, subsequently molding whichever of the substrate and the support member has not already been molded on the initially molded component may be implemented prior to the initially molded component returning to room temperature, and preferably while a temperature of the initially molded component is at least 80°C. This may assist adhesion of the substrate and the support member to each other. In embodiments, in which the subsequent molding comprises curing the precursor material, such maintenance of elevated temperature can also assist such curing.

[0028] In some embodiments, the method comprises arranging at least part of a base member in or adjacent to the mold, and initially molding the substrate or the support member on the base member.

[0029] Following molding of whichever of the substrate and the support member has not already been molded on the initially molded component, the base member with the substrate and the support member molded thereon may be removed, e.g., released, from the mold.

[0030] The base member may assist to reinforce the device. Alternatively, or additionally, the base member, e.g. the glass base member, may be optically transparent to enable analysis of the biological material through the glass base member via optical microscopy.

[0031] The base member, e.g., glass base member, may also provide a smooth surface on which to mold the support member. Such a smooth surface may, in turn, assist an exterior surface of the support member to have a smooth surface. This may facilitate analysis of the biological material through the support member via optical microscopy.

[0032] It is noted that in other embodiments in which the base member is not included in the device, or more generally the exterior surface of the support member is in contact with a surface of the mold during molding, the surface of the mold is preferably a polished surface. Such a polished surface may assist to provide a relatively smooth exterior surface of the support member, which can ultimately facilitate analysis of the biological material through the support member via optical microscopy.

[0033] In some embodiments, the method comprises releasing the substrate together with the support member from the mold while temperatures of the substrate and the support member are 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.

[0034] In some embodiments, the method comprises adjusting the mold from a first configuration used for molding said one of the substrate and the support member to a second configuration used for molding said other of the substrate and the support member. In such embodiments, the adjusting the mold from the first configuration to the second configuration may comprise switching or replacing a second mold part with a third mold part while the component initially molded between a first mold part and the second mold part remains received in the first mold part. Such switching may, for example, include opening the mold by displacing the second mold part, followed by re-closing the mold using the third mold part.

[0035] In preferred embodiments, the method preferably comprises the mold including a second mold configuration for molding the support member onto a substrate, the second mold configuration comprising a second mold part and a third mold part separable from the first mold part and that together form the second mold configuration. In some embodiments the substrate may have been previously molded and is 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 the first mold part and a second mold part, the second mold part being separable from the first mold part and from the substrate once molded such that the substrate is left within the first mold part. In such embodiments, the adjusting the mold from the first configuration to the second configuration may thus comprise switching or replacing a 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 may, for example, include opening the mold by displacing the second mold part, followed by re-closing the mold using the third mold part.

[0036] In preferred embodiments using the second mold configuration, and optionally but preferably also the first mold configuration, the support member is molded on the substrate. Preferably the substrate comprises, or even consists of, a thermoplastic or thermoset material, either one not being a rubber. Preferably such material is transparent. For example, and preferably the material comprises at least one component chosen from the group consisting of: polycarbonate, polyether ether ketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and polyethersulfone (PES). The most preferred material comprises polycarbonate. Such materials generally have one or more of the following desired properties: thermal properties as further described herein, transparent to visible light, stiffness (e.g. not a rubber) and availability / cost. A substrate comprising or consisting of any one of these materials preferably is combined with a support member comprising a rubber material. A preferred rubber material comprises silicone, such as one or more silicone polymers as described herein. Preferably silicone rubber material is molded using a precursor material as described herein that undergoes a curing as described herein. Most preferably, the silicone rubber material is a silicone rubber material modified with polar groups such as for example carboxylic acid groups or their conjugated base groups.

[0037] In some embodiments, the support member may be formed from a softer, e.g., lower Shore A hardness, and / or more flexible material than that forming the substrate. Thus, the mechanical properties of the support member may be appropriate for supporting the biological material, e.g., cells or tissue being cultured in the chamber unit(s), while the substrate contributes more to the device’s structural rigidity.

[0038] Alternatively, or additionally, the support member may be formed from a more biocompatible material than that forming the substrate so that cells and / or tissue preferentially adhere(s) to the support member. In this way, selection of materials can be used to guide cell / tissue growth where such cell / tissue growth is intended, e.g., in region(s) of the device that facilitate observation and / or testing of the cells / tissue.

[0039] In some embodiments, the support member comprises silicone. Preferably, the silicone comprises or even consists of a silicone polymer in the form of a rubber. Such a silicone support member may be optically transparent and also have limited autofluorescence so that the support member may allow a variety of conventionally used optical inspection techniques and protocols, e.g., with or without staining, to be used in testing performed using the device. Moreover, silicone may be suitably biocompatible, particularly when modified so that polar groups are available at the biological materialcontacting surface(s) of the silicone substrate.

[0040] In more general terms, the support member may comprise a polymeric material that is bulk-modified with moieties that each include a polar group, with polar groups of the moieties being available at a surface of the support member arranged to contact the biological material received in the at least one chamber unit. Such polar groups may assist to render the support member biocompatible, e.g., via application of a protein such as fibronectin to the polar group-functionalized surface of the support member.

[0041] In some embodiments, the substrate is formed from a material having a glass transition temperature of at least 140°C. This minimum glass transition temperature may permit, for example, molding of the support member on the substate, e.g., with such molding comprising relatively high temperature curing of a precursor material.

[0042] Alternatively, or additionally, the substrate may be formed from a thermoplastic, preferably a thermoplastic selected from one or more of polycarbonate, polyether ether ketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and polyethersulfone (PES).

[0043] In some embodiments, the substrate provides sidewall(s) of the at least one chamber unit, with a central portion of the support member being provided in the at least one chamber unit, and with a plurality of rib elements each extending from the central portion to engage with the respective chamber unit’s sidewall. Such rib elements may assist to minimize deformation of the central portion while keeping shear stresses sufficiently high to assist to keep the support member in position.

[0044] In some embodiments, the support member comprises at least one structural feature for contacting the biological material received in the at least one chamber unit. Such structural feature(s) may provide a suitable shape or profile for growth of cells or tissue thereon. The shape or profile of the structural feature(s) may be selected according to, for example, the type of cells or tissue being grown and / or the testing to be carried out on the biological material. The structural features can be chosen from the group consisting of: a membrane; a porous membrane; a plurality of wells each having a membrane or porous membrane and a membrane having an opening or slit for clamping biological material such as a biopsy. Other structural features may however be chosen.

[0045] In some embodiments, the at least one structural feature comprises, per chamber unit, a pair of flexible protrusions, e.g., flexible posts or pillars, for supporting tissue thereon and therebetween. In such embodiments, the device may be employed 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 may deflect, with displacement of the protrusions caused by the deflection being measurable to enable determination of a force provided by contraction of the tissue.

[0046] In some embodiments, the at least one chamber unit comprises a plurality of chamber units. Thus, testing of biological material can be implemented in different chamber units at the same time. In such embodiments, the chamber units can be arranged in one or more rows, e.g., in a manner akin to a conventional well plate.

[0047] 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 arranged to hold one of a substrate and support member such that in the second mold configuration the first mold part, the second mold part and the one of the substrate and support member enclose one or more second open spaces in which the other of the substrate and support member can be molded onto the one of the substrate and support member. In preferred embodiments one of the substrate and support member consists of the substrate and the other of the substrate and support member consists of the support member. In such embodiments, the second mold configuration is configured for molding the support member onto the substrate.

[0048] Preferably, the first mold part comprises at least one first mold surface for defining a chamber surface of an end portion. The first mold surface can thus 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 which includes 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) at least part of a 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. The first mold surface can thus be shaped to provide the end-portion of a support member with a desired design, such as pillars, membrane, wells etc. as defined herein. The first mold surface can have any shape desired as described herein such as flat, curved, having protrusions or indentations or recesses according to the desired shape of the support member to be formed against that surface.

[0049] These mold designs can be used to make chamber-units that have some of their wall defined by the substrate and some of their wall defined by the support member such that in the chamber there is at least some surface of the support member exposed for the support of the biological material.

[0050] The first mold part is further arranged such that, if it holds a substrate, at least one, preferably a plurality, of contact surfaces of the substrate are exposed within the one or more spaces of the second mold configuration. Therewith, the support member can be molded onto these contact surfaces using the second mold configuration.

[0051] The first mold part can comprise one or more protrusions for occupying (e.g. filling) channels in a substate.

[0052] Preferably, the third mold part comprises at least one third mold surface for defining an end portion. The first mold surface can thus be used to design the end portion surface that will be used to support biological material in a finished device. The third mold surface may be arranged to define one or more of: a flange, membrane or pillar structure. The first mold surface and third mold surface preferably are arranged in the respective mold parts such that in the second mold configuration they define part of the one or more second open spaces in which an end-portion of the support member is defined. Preferably the third mold surface is comprised in a protrusion arranged in the third mold part such that it extends towards the first mold surface in the one or more open second openings of the second mold configuration. Preferably the protrusion defines a sub-chamber in the support member. Preferably the third mold 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 connect to the one or more subchambers.

[0053] Preferably the mold system further comprises a second mold part separable from the first mold part and designed and arranged to be combined with the first mold part to form a first mold configuration in which the first mold part and the second mold part enclose one or more first open spaces in which the one of the substrate and support member can be molded. While this second mold part is not needed in a method that only uses the second mold configuration. For example, when preformed substrates are used in the process, in methods that include molding of the substrate, this second mold part is of use and advantage.

[0054] Preferably the second mold part comprises a second mold surface for defining the contact surface of a substrate to be molded. It is this second mold surface that can be shaped to improve adhesion of the support member to the substrate. Preferably the second contact surface comprises at least one of: corrugations, indentations, protrusions and surface roughness. Any shapes that increase the area of the contact surface with respect to that of a flat contact surface is beneficial for adhesion.

[0055] Preferably, the second mold part comprises a further chamber protrusion arranged to extend in the one or more first open spaces of the first mold configuration such that in the first mold configuration the combination of the chamber protrusion of the first mold part and the further chamber protrusion defines a chamber unit in a substrate.

[0056] The mold system preferably is a mold system for injection molding. Preferably one or more of the mold parts comprise one or more openings for injection of materials to be molded into the substrate and support member. Preferably the mold parts are made of metals such as, for example, steel.

[0057] Preferably, the shape and size of the first mold part, the second mold part and the third mold part are such that a device, and preferably the substrate of such device, includes at least one, and preferably a plurality of, chambers.

[0058] According to another aspect there is provided a biological material testing device comprising: a substrate in which at least one chamber unit is defined; and a support member for supporting biological material received in the at least one chamber unit, one of the substrate and the support member being molded on the other of the substrate and the support member.

[0059] The device may be obtainable by the method according to any of the embodiments described herein. The device may be obtainable by molding one of the substrate and the support member on the other of the substrate and the support member.

[0060] The device preferably comprises at least one chamber, wherein the at least one chamber is defined by part of the walls of the chamber unit and by part of a surface of the support member. The part 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 unit such that it forms the bottom of the chamber, or at least part of the bottom of the chamber.

[0061] The chamber-unit can have one chamber, or a plurality of chambers. The device can have a plurality of chambers. These may be arranged in one or more arrays. The device, in some embodiments, comprises fluid channels from and to chambers. Preferably these channels are arranged in the substrate. The chambers, in some embodiments, are arranged to connect multiple chambers. Alternatively, or additionally, the support member comprises at least one, such as a plurality, of fluid channels. The device in some embodiments comprises at least one membrane, where 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 well. In some embodiments, the well comprises the membrane. The end portion can comprise a flange that extends at least partly into a chamber unit. The flange is arranged to carry, or be integrated with, the membrane or with a part of the end portion comprising a well as defined herein. Preferably the end-portion comprises the porous membrane and the flange and the support member is further arranged to define a sub-chamber that is bounded by the membrane and that is connected to one or more fluid channels arranged in the support member.

[0062] Embodiments described herein in relation to the method may be applicable to the device, and embodiments described herein in relation to the device may be applicable to the method. Thus, preferred materials described for the methods are also preferred materials for the devices per se. Similarly, preferred structural device features of the devices as described in relation to the methods disclosed and defined herein are also preferred features for the devices per se.

[0063] According to yet another aspect there is provided use of the device according to any of the embodiments described herein for testing biological material.

[0064] According to a further aspect there is provided a drug testing method 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 the drug to be tested.

[0065] In some embodiments, the providing of biological material in the at least one chamber unit comprises culturing cells in the at least one chamber unit.

[0066] Alternatively, or additionally, the drug testing method may comprise monitoring a response of the biological material, e.g., cultured cells, to the drug to be tested. This monitoring may be implemented in any suitable manner, for example by optical microscopy, e.g., through the support member when the support member is optically transparent.

[0067] These and other aspects of the disclosure will be apparent from and elucidated with reference to the embodiment(s) described hereinafter.

[0068] BRIEF DESCRIPTION OF THE DRAWINGS

[0069] For a better understanding of the disclosure , and to show more clearly how it may be carried into effect, reference will now be made, by way of example only, to the accompanying drawings, in which:

[0070] FIGs. 1A and IB schematically depict a method for manufacturing a biological material testing device according to a first example;

[0071] FIG. 2 A shows part of an exemplary support member for inclusion in the device;

[0072] FIG. 2B provides views of another exemplary support member for inclusion in the device;

[0073] FIG. 3 provides a flowchart of a method for manufacturing a biological material testing device according to another example;

[0074] FIG. 4 shows a device according to a second example;

[0075] FIGs. 5A to 5C provide various views of a device according to a third example;

[0076] FIG. 6 shows a device according to a fourth example;

[0077] FIG. 7 shows a device according to a fifth example; and FIG. 8 shows a device according to sixth example. FIG. 9A shows a set of mold parts to form a mold of a first mold configuration to be used for molding a substrate of the device of Figs. 9E to 9H using methods disclosed herein;

[0078] Fig. 9B shows the first mold configuration using the mold parts of Fig. 9A with the substrate of the device of Figs. 9E to 9H molded therein;

[0079] Figs.9C and 9D show replacing a mold part to provide a second mold configuration for molding the support member of the device of Figs. 9E to 9H according to methods disclosed herein. The second mold configuration and the support member molded therein are shown in Fig. 9D;

[0080] Figs 9E to 9H show cross sections as well as top and bottom views of a device molded according to methods disclosed herein.

[0081] DETAILED DESCRIPTION OF THE EMBODIMENTS

[0082] The disclosure will be described with reference to the Figures.

[0083] The detailed description and specific examples, while indicating exemplary embodiments of the apparatus, systems and methods, are intended for purposes of illustration only and are not intended to limit the scope of the disclosure. These and other features, aspects, and advantages of the apparatus, systems and methods of the present disclosure will become better understood from the following description, appended claims, and accompanying drawings. The Figures are merely schematic and are not drawn to scale. It should also be understood that the same reference numerals are used throughout the Figures to indicate the same or similar parts.

[0084] Provided is a biological material testing device comprising a substrate in which at least one chamber unit is defined, and a support member for supporting biological material received 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. Also provided is a method for manufacturing such a biological material testing device. Further provided are uses and methods in which the device is employed for testing biological material, for example in drug testing in which the biological material is exposed to a drug to be tested.

[0085] FIGs. 1A and IB schematically depict a device design and a manufacturing method 10 according to an example according to the disclosure. Method 10 is for manufacturing a device 100 for testing biological material. To this end, device 100 comprises a substrate 102 in which at least one chamber unit 104 is defined. Substrate 102 shown in FIGs. 1 A and IB has a plurality, in this case 8, of such chamber units 104. The device 100 further comprises a support member 106 for supporting biological material (the biological material is not visible in FIGs. 1A and IB) when such biological material is received in the, e.g., each of, the chamber unit(s) 104.

[0086] Examples of biological material include cell groups, double cell layers, spheroids, organoids or biopsies, but others may be used too.

[0087] Each chamber unit 104 may have a height extending between a top side and the 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 may be in the range of 3 to 20 mm, such as 3 to 10 mm, e.g., about 6 mm.

[0088] Such dimensions may balance the requirement for the chamber unit(s) 104 to be kept as small as possible, e.g., to enable as many chamber unit(s) 104 to be defined in the substrate 102 as possible, with the requirement for the chamber unit(s) 104 to provide sufficient space for testing of the biological material, e.g., for capture and subsequent analysis of cells.

[0089] The chamber unit(s) 104 may have any suitable shape. A generally cylindrical shape, such as shown in the FIGs 1A and IB, may be preferred from a fluid dynamics perspective, e.g., to mitigate the risk of pooling of fluid in a corner of a chamber unit 104 and / or to ensure usability with existing analysis equipment. However, shapes other than cylindrical can be contemplated, e.g., shapes having a square or rectangular cross-section.

[0090] In some embodiments, such as shown in FIGs. 1A and IB, the substrate 102 provides sidewall(s) 108, e.g., cylindrical sidewall(s) 108, of the at least one chamber unit 104, and the support member 106 provides an end portion 110 from which the sidewall(s) 108 extend(s). The end portion 110 in combination with the sidewall(s) 108 of each chamber unit 104 may provide a container in which the biological material can reside, e.g., together with cell culture medium. Typically end-portions 110 can be referred to as the bottom of a chamber where a chamber may also be referred to as a container.

[0091] In general, each chamber or container has a chamber depth measured along the sidewall from the top side of the substrate to the end-portion. In the example of Fig. 1 A to IB, the height is equal to the chamber depth, but this need not be the case. In variations of the example, as will be disclosed herein, the support member may extend into a chamber unit from the bottom side of the chamber unit such that the chamber depth is smaller than the height by the distance the support member extends into the chamber unit form the bottom side.

[0092] The thickness of the support member 106, e.g., the end portion(s) 110 provided by the support member 106, may be selected to enable analysis of the biological material received in the chamber unit(s) 104 through the support member 106.

[0093] Such analysis may comprise 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, may be optically transparent, and may have a thickness that is, for example, at most 0.4 mm. Optical in this sense can pertain to the spectral region comprising one, more or all of visible light, UV light and Near Infrared Light.

[0094] 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 device. Thus, testing of biological material can be implemented in different chamber units 104 at the same time. In such embodiments, the chamber units 104 can be arranged in one or more rows 105. One such row 105, comprising eight chamber units 104, is shown in FIGs. 1A and IB, although more than one row 105, e.g., with each of the rows 105 comprising eight, fewer than eight or greater than eight chamber units 104 can also be contemplated, as evident from further examples explained herein below.

[0095] In some embodiments, such as shown in FIGs. 1A and IB, the device 100, e.g., the substrate 102 thereof, comprises one or more mounting features 111, such as tabs, to enable the device 100 to be mounted on and / or in a holder (not visible). In such embodiments, the holder may, for instance, be configured to simultaneously support a plurality of such devices 100 thereon and / or therein.

[0096] The holder may, for example, hold twelve of the devices 100 shown in FIGs. 1A and IB, such that the holder in combination with the twelve devices 100 defines a ninety-six chamber unit 104 device 100, e.g., a ninety-six well plate.

[0097] In some embodiments, one or more of the devices 100 is or are individually detachable from the holder.

[0098] This may provide a more flexible holder-device(s) arrangement. In relation to the nonlimiting example in which twelve of the devices 100 are mountable in the holder, detachment of one or more of the devices 100 may be implemented when the experimentalist does not wish to use all ninety-six chamber units 104 at once.

[0099] Biological material tends to be required to be living during at least part of the testing performed using the device 100. Hence the device 100, for example at least the support member 106 thereof, may be formed from a suitable biocompatible material. In some embodiments, the support member 106 is formed from a biocompatible polymeric material, such as silicone, e.g., to which a suitable cell culture protein, such as fibronectin, may be, or is applied. Suitable materials are mentioned herein.

[0100] In some embodiments, the support member 106 comprises a polymeric material that is bulk-modified with moieties that each include a polar group, with polar groups of the moieties being available at a surface of the support member 106 arranged to contact the biological material received in the at least one chamber unit 104.

[0101] Such polar groups may assist to render the support member 106 biocompatible, e.g., via application of a protein such as fibronectin to the polar group-functionalized surface of the support member.

[0102] In some embodiments, the moieties comprise fatty acid moieties, with the polar groups comprising carboxylic acid groups of the fatty acid moieties.

[0103] Particular mention is made of silicone being bulk-modified with polar group-comprising moieties, e.g., fatty acids, for instance via crosslinking between silicon-hydrogen bonds and C=C bonds of unsaturated fatty acid moieties.

[0104] This crosslinking may be catalyzed by a platinum catalyst.

[0105] In this connection, reference is made to materials and methods described, for example, in WO2021058657 and WO2019015988 which further describe the processing of preferred materials to be used for manufacture of a support member as defined herein and are incorporated by reference in their entirety herein. Thus, with reference to the documents, 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 conjugated bases are preferred over sulphonic acid groups and carboxylic acid groups preferred over phosphoric acid groups. Residues carrying such groups are more easy to use during the bulk modification as during such modification method their precursors having the acidic groups in conjugated base form with metal counter ions and such residues mix better with the other consitiuents to form the elastomer from. The acidic groups may be chosen according to pKa (as specified herein before) needed.

[0106] In some preferred embodiments the residues comprise or consist of an aliphatic portion comprising 3 or more carbon atomes and less than 50 carbon atoms, the one or more acidic groups being covalently coupled to the aliphatic portion and the aliphatic portion being covalently coupled to the elatomer bulk. The aliphatic portion may be linear or branched. It can comprise one or more carboncarbon double or triple bonds or aryl or benzene units. The aliphatic portion can comprise cyclic units such as cyclohexyl or cyclopentyl or other. The aliphatic portion prefearalby is a saturated hydrocarbon portion. The aliphatic portion preferably comprises only carbon and hydrogen atoms.

[0107] In some embodiments the aliphatic portion is linear chain coupled to the elastomer bulk at a terminal end. In some embodiments the aliphatic portion further comprises no carbon-carbon triple bonds to increase flexibility of the residue.

[0108] In some embodiments the residues within a bulk modified elastomer may be different from each other each one chosen as defined herein. In some embodiments at least part of the residues maybe bound to the elastomer bulk via two covalent bonds. This may for example result from residual precursors that have two ethylenic bonds for participating in the modification reaction. In some embodiments the portion comprise at least 5 carbon atoms and more preferably at least 10 carbon atoms. Preferably the aliphatic portion comprises less than 40 or less than 30 carbon atoms. Linear portions are preferred, but this is not needed perse. The number of carbon atoms of such portion is preferably between 5 and 30 more preferably between 5 and 20 or 5 and 15. The portion and, or chain may have one or more aryl groups. One or more of the acidic groups may be directly attached to an aryl group such as a benzene ring.

[0109] In some embodiments the residues are the remainder of unsaturated fatty acid precursors that were covalently bound to the elastomer bulk via reaction of one or more (if present) of their ethylenic groups (carbon-carbon double bonds).

[0110] For example, the unsaurated fatty acid residues are one or more residues of fatty acids chosen from the group consisting of: myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoeladic acid, a-linolenic acid, arachidonic acid, eicospaentaenoic acid, erucic acid and docosahexaenoic acid

[0111] In some embodiments the residues are the remainders of precursors having linear or branched alkyl chains 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 may provide increased reactivity as compared to non-terminal ethylenic groups during formation of the elastomer bulk in the manufacturing process. Linear chains with one terminal ethylenic group and one acidic group such as e.g. a carboxyl group are preferred examples. In such case there may be between 5 and 15 carbon atoms in the chain.

[0112] In some embodiments the elastomer bulk comprises a silicone or a poly diene backbone. Polybutadiene and polyisorpene are examples of polydienes. Polydimethylsiloxane is an example of a polysioxane (silicone). The silicones have a higher water permeability and are more transparent allowing easier optical inspection of cell culturing.

[0113] In some embodiments the residues are covalently bound to the elastomer bulk as a result of a reaction between an unsaturated carbon-carbon bond of a precursor of the residue and a vinyl or hydride functional group of the elastomer bulk. In poly dienes such reaction is to vinyl groups while in silicones such binding is usually with silylhydirde functional groups.

[0114] More generally, the term “biocompatible” as used herein may refer to a material capable of allowing cells to proliferate on the material.

[0115] The term “more biocompatible material” may refer to a material for which a degree of cell proliferation thereon is higher than a given material to which the more biocompatible material is being compared.

[0116] In some embodiments, the support member 106 is formed from a more biocompatible material than that forming the substrate 102 so that cells and / or tissue preferentially adhere(s) to the support member 106.

[0117] For example, silicone, e.g., silicone modified in the manner described above, may form the support member 106, whereas a polymeric material for which the biological material has less affinity, such as a polymeric material comprising one or more materials chosen from the group consisting of polycarbonate, polyether ether ketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and poly ethersulfone (PES), may form at least part of the substrate 102. For instance, a polymeric material for which the biological material has less affinity, such as polycarbonate, polyether ether ketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and / or polyethersulfone (PES) may form the sidewall(s) 108 of the chamber unit(s) 104.

[0118] In at least some embodiments, the support member 106 comprises an elastomeric material (sometimes herein also referred to as a rubber or rubber material), such as an elastomeric material comprising one or more materials chosen from the group consisting of silicone and polybutadiene. The elastic properties of such an elastomeric material, e.g., together with such a material being biocompatible or at least being modifiable to provide sufficient biocompatibility, may make such an elastomeric material particularly appropriate for inclusion in the support member 106.

[0119] Alternatively, or additionally, the support member 106 may be formed from a softer, e.g., lower Shore A hardness, and / or more flexible material than that forming the substrate 102. Thus, the mechanical properties of the support member 106 may be appropriate for supporting the biological material, e.g., cells or tissue being cultured in the chamber unit(s) 104, while the substrate 102 contributes more to the device’s 100 structural rigidity.

[0120] It is also noted that embodiments in which the support member 106 comprises, e.g., is formed from, silicone, the silicone support member 106 may be optically transparent and also have limited autofluorescence so that the support member 106 may allow a variety of conventionally used optical inspection techniques and protocols, e.g., with or without staining, to be used in testing performed using the device 100.

[0121] Selection of materials for the substrate 102 and the support member 106 is further discussed in more detail herein below in relation to method 10 of manufacturing the device 100.

[0122] A list of exemplary materials for the support member 106 is provided below.

[0123] In some embodiments, and referring again to FIGs. 1A and IB, the support member 106 comprises at least one structural feature 112 for contacting the biological material received in the chamber unit(s) 104. Such a structural feature 112 may provide a suitable shape or profile for growth of cells or tissue thereon. The shape or profile of the structural feature 112 may be selected according to, for example, the type of cells or tissue being grown and / or the testing to be carried out on the biological material.

[0124] In some embodiments, such as shown in FIGs. 1A, IB, 2 A and 2B, the at least one structural feature 112 comprises, per chamber unit 104, a pair of flexible protrusions, e.g., flexible posts or pillars, for supporting tissue thereon and therebetween. In such embodiments, the device 100 may be employed 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 may deflect to bend towards each other, with displacement of the protrusions caused by the deflection being measurable to enable determination of one or more contraction properties such as for example an extent of contraction, a frequency of contraction and / or a force of contraction provided by contraction of the tissue.

[0125] Such measurement is described, for example, in WO 2022 / 112291, which is incorporated herein by reference in its entirety.

[0126] The protrusions may have any suitable height provided that they are able to fulfil their tissue testing function. In some embodiments, and referring to FIG. 2B, the protrusions each have a height H or largest dimension that is 500 to 1500 pm, preferably 600 to 1000 pm.

[0127] Such a height H, e.g., together with the thickness of support member 106 between an exterior surface of the support member 106 to the interior surface 116 from which the protrusions protrude being at most 4 mm, may assist with analysis of the tissue by optical microscopy through the support member 106 when the support member 106 is optically transparent at least beneath the protrusions.

[0128] Alternatively, or additionally, the protrusions may each have a width W of 100 to 300 pm and / or a length L of 400 to 600 pm. A spacing SP between the protrusions, e.g., protrusions having the above height H, width W and / or length L dimensions, may be 800 to 1200 pm.

[0129] It is noted that the flexibility of the protrusions may be provided by at least the protrusions of the support member 106 comprising an elastomeric material, such as an elastomeric material comprising one or more materials chosen from the group consisting of silicone and polybutadiene.

[0130] In some embodiments, and as best shown in FIGs. 2A and 2B, a recess 114 per chamber unit 104 is defined in the support member 106. A base surface 116 that partly delimits the recess 114 may be joined to a main surface 118 of the support member 106 by a side surface 120 that extends between the base surface 116 and the main surface 118.

[0131] The structural feature(s) 112, e.g., flexible protrusions, may be arranged on and / or in the base surface 116 of the recess 114. Alternatively, or additionally, the recess 114 may be elongated, e.g., ellipsoidal, when viewed in plan, as shown in FIG. IB and the upper pane of FIG. 2B.

[0132] In the case of such an elongated recess 114, and when the protrusions are included in the support member 106, the protrusions may be arranged along a longitudinal axis 122 along which the recess 114 is elongated. Contraction of muscle tissue between the protrusions may be along this longitudinal axis 122.

[0133] The recess 114, e.g., elongated recess 114, can have any suitable dimensions. In some embodiments, and referring to FIG. 2B, the recess 114 has a width W1 of 1500 to 2500 pm, preferably 1800 to 2200 pm, and / or a length LI of 2500 to 3500 pm, preferably 2700 to 3200 pm. Plainly the length LI, extending along the longitudinal axis 122, is larger than the width W1 in embodiments in which the recess 114 is elongated.

[0134] The height Hl of the recess 114, e.g., the recess 114 having the above width W1 and / or length LI dimensions, may be 500 to 1500 pm, preferably 600 to 1000 pm.

[0135] It is noted that the height Hl of the recess 114 may be the same, or within 10%, of the height H of the protrusions when present.

[0136] It is noted that in some embodiments, the structural feature(s) 112, e.g., protrusions, are included in the support member 106, but without any recess 114 being defined in the support member 106.

[0137] In such embodiments, the sidewall(s) 108 of the chamber unit(s) 104 may be relied upon to provide suitable lateral confinement of the biological material, cell culture medium, etc. within each of the chamber unit(s) 104.

[0138] In some embodiments, such as shown in FIG. 2 A, a central portion 124 of the support member 106 is provided in the, e.g., each of the, at least one chamber unit 104, with a plurality of rib elements 126 each extending from the central portion 124 to engage with the respective chamber unit’s 104 sidewall 108. Such rib elements 126 may assist to reduce or minimize deformation of the central portion 124 while keeping shear stresses sufficiently high to assist to keep the support member 106 in position.

[0139] Any suitable number of rib elements 126 can be contemplated, such as two, three, four (as shown in FIG. 2A), five or more. Preferably, a symmetrical arrangement of rib elements 126 is employed, with two rib elements 126 being arranged along a first axis 128 and two rib elements 126 being arranged along a second axis 130 perpendicular to the first axis 128.

[0140] It is noted that recess 114 may be defined in the central portion 124 of the support member 106, for example with structural feature(s) 112, e.g., flexible protrusions, being arranged on and / or in the base surface 116 of the recess 114. In the case of an elongated recess 114, the above- mentioned longitudinal axis 122 may coincide with an axis 128 along which rib elements 126 extend to reach the chamber unit’s 104 sidewall 108.

[0141] 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, for example by forming apertures in the material, e.g., elastomeric material, constituting the support member 106. Such apertures can be formed, for example, by laser ablation or casting imprint perforation.

[0142] Laser perforation / ablation is described, for example, in US2014127744 and US20180315409. Casting imprint perforation is described, for instance, in US2020360923, US2015010919 and US2022228108 all of which are incorporated by reference herein in their entirety.

[0143] Each of the apertures is preferably sized to restrict or prevent cells supported on the support member 106 from passing therethrough. To this end, each aperture may have a diameter less than 10 pm.

[0144] Generally, attaching the substrate 102 and the support member 106 of different materials as disclosed herein, e.g. of a device as described with reference to Figs 1A to ID and 2A and 2B, to each other may present various technical challenges. It may be difficult to implement such attachment in a sterile manner and / or without inadvertently providing points between the substrate 102 and the support member 106 where fluid leakage can occur potentially hampering use. Moreover, additional manufacturing steps may be necessary to attach substrate 102 and the support member 106 to each other increasing manufacturing complexity and cost. Furthermore, In embodiments in which the support member 106 includes the flexible protrusions, the substrate 102 and the support member 106 may need to be aligned to ensure that the protrusions are correctly orientated, e.g., in each chamber unit 108 of a plurality of chamber units 108.

[0145] For at least these reasons, and referring again to FIGs. 1 A and IB, the method 10 according to the disclosure comprises molding 12, using a mold, one of the substrate and the support member 102, 106 on at least part of the other of the substrate and the support member 106, 102, which at least part of the other of the substrate and the support member 106, 102 is arranged in or adjacent to the mold during the molding 12. For example, in the embodiment schematically depicted in FIGs. 1A and IB, the support member 106 is molded onto the substrate 102.

[0146] By molding 12 one of the substrate and the support member 102, 106 on at least part of the other of the substrate and the support member 106, 102, the former may be shaped and adhered onto the latter at the same time. 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 may be obviated. In particular, no adhesive / glue may be required to keep the substrate 102 and the support member 106 attached to each other.

[0147] The method 10 may provide a relatively quick and reliable process for manufacturing the device 100, in which the substrate 102 and the support member 106 can be regarded as being integrated with each other in a single piece. Moreover, the support member 106, e.g. silicone support member 106, may be anchored by geometry into a frame, e.g. polycarbonate frame, defined by the substrate 102. Thus, parts of the substrate may be embedded in parts of the support member such that one cannot be easily separated from the other.

[0148] Accordingly, the adhesion, e.g. leakage tight adhesion, provided by the molding 12 of one of the substrate and the support member 102, 106 on the other of the substrate and the support member 106, 102 may be supplemented by geometrical anchoring, particularly in embodiments in which the support member 106 comprises the central portion 124 and the rib elements 126 that engage the sidewall(s) 108 of the chamber unit(s) 104. In other embodiments, the central portion-rib elements arrangement 124, 126 may be omitted, for example to simplify the design of the mold.

[0149] It is noted also that on a microscopic scale the irregularities (e.g. molecular scale caused irregularities) of a surface of the first molded part (e.g. the substrate 102) may be conformed to, at least in part, by the second molded part (e.g. the support member 106) due to its molding onto the first molded part. This may add to improving adhesion of both parts of a device disclosed herein that has been made using methods disclosed herein and benefit its leakage characteristics. The method in which one part is molded onto the other part may thus allow use of irregularly shaped contact surfaces between a substrate and support member that otherwise would be more difficult to bring in good contact. For example, separately made components with contact surfaces having complementary and matching complex irregular shapes for intimate contact may be difficult or more expensive to make. Inspection of a contact surface between substrate and support member of a device as disclosed herein may thus be used to distinguish a device as disclosed herein from other multicomponent devices not made using the methods as disclosed herein such as for example methods making use of clamping, adhering or glueing of separately made parts. Although conformal adherence may also be observed for parts that are glued together, here the additional glue layer may help to distinguish such glued device from one made using the methods disclosed herein.

[0150] Implicit in the molding 12 is that the one of the substrate and the support member 102,

[0151] 106 on which the other of the substrate and the support member 106, 102 is molded can withstand the molding conditions. This may influence one or more of the following: which of the substrate 102 and the support member 106 is molded 12 onto the other (and conversely which component is already fabricated prior to the molding 12); the materials and / or precursors selected for forming the substrate 102 and the support member 106; and the conditions during the molding 12.

[0152] In at least some embodiments, the molding 12 comprises, e.g., is defined by, injection molding the one of the substrate and the support member 102, 106 on the at least part of the other of the substrate and the support member 106, 102. Injection molding may provide a cost-effective and scalable way of manufacturing the device 100.

[0153] In some embodiments, such as shown in FIG. 3, the method 10 comprises releasing 14 the substrate 102 together with the support member 106 from the mold while temperatures of the substrate 102 and the support member 106 are 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.

[0154] In some embodiments, and still referring to FIG. 3, the molding 12, e.g., injection molding, of the one of the substrate and the support member 102, 106 on the at least part of the other of the substrate and the support member 106, 102 comprises curing 12B a precursor material. In the case of injection molding, the molding 12 may comprise 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 the support member 106, 102.

[0155] In such embodiments, the curing 12B may comprise heating the precursor material, for example above 160°C, e.g., between 160 and 200°C.

[0156] Heating at such temperatures may assist to ensure that the precursor material is cured and adhered to the initially molded component.

[0157] Following the heating, the substrate 102 and the support member 106 may be cooled to room temperature, albeit with the substrate 102 together with the support member 106 being preferably released 14 from the mold prior to the substrate 102 and the support member 106 cooling to room temperature, as previously described.

[0158] It is noted that thermal curing 12B of the precursor material may require selection of a suitable thermally robust material for the one of the substrate and the support member 102, 106 on which the other of the substrate and the support member 106, 102 is molded via curing 12B.

[0159] To this end, a glass transition temperature of the material forming the one of the substrate and the support member 102, 106 on which the other of the substrate and the support member 106, 102 is molded 12 may be at least 140°C.

[0160] This minimum glass transition temperature may permit relatively high temperature curing 12B of the precursor material during the molding 12.

[0161] 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), as well as combinations of the above polymers, e.g., blends or alloys of at least two of poly ether ether ketone (PEEK), polyetherimide (PEI) and polyethersulfone (PES).

[0162] In embodiments in which the support member 106 comprises, e.g., is formed from silicone, curing 12B the precursor material may comprise curing a siloxane precursor material.

[0163] Such curing 12B may be implemented, for example, between 140°C and 180°C. Alternatively, or additionally, the precursor material, e.g. the siloxane precursor material, may be injected into the mold at a pressure between 100 and 600 bar, preferably when the mold is above 110°C.

[0164] In some embodiments, such as shown in FIGs. 1A, IB and 3, the method 10 comprises initially molding 16, e.g., initially injection molding 16, the substrate 102 or the support member 106, and subsequently molding 12; 12A, 12B, e.g., subsequently injection molding 12; 12A, 12B, whichever of the substrate 102 and the support member 106 has not already been molded on the initially molded component. Thus, the method 10 may comprise a two-step molding process, for example a two-step injection molding process.

[0165] The initial molding 16 may involve injection molding 16 a thermoplastic, for example a thermoplastic selected from one or more of polycarbonate, poly ether ether ketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and polyethersulfone (PES). Such a thermoplastic may, for instance, be melted, e.g., at a temperature of 200 to 300°C, injected at a pressure between 600 and 2000 bar into the mold that is preferably at a temperature less than the glass transition temperature of the thermoplastic. For example, the temperature of the mold may be 80 to 120°C.

[0166] In some embodiments, and referring to FIG. 3, the method 10 comprises adjusting 18A, 18B the mold from a first configuration used for initially molding 16, e.g. initially injection molding 16, the substrate 102 or the support member 106, to a second configuration used for subsequently molding 12; 12A, 12B, e.g., subsequently injection molding 12; 12A, 12B, whichever of the substrate 102 and the support member 106 has not already been molded on the initially molded component.

[0167] The mold may include a first mold part and a second mold part, with the substrate 102 or the support member 106 being moldable between the first mold part and the second mold part. The mold may further comprise a third mold part, with whichever of the substrate 102 and the support member 106 that is not moldable between the first mold part and the second mold part being moldable between the third part and the component that has been molded between the first mold part and the second mold part.

[0168] In such embodiments, the adjusting 18 A, 18B the mold from the first configuration to the second configuration may comprise switching the second mold part with the third mold part while the component molded between the first mold part and the second mold part remains received in the first mold part. Such switching may, for example, include opening 18A the mold by displacing the second mold part, followed by re-closing 18B the mold using the third mold part.

[0169] Initially molding 16, e.g., injection molding 16, the substrate 102 or the support member 106 may comprise heating a material above room temperature to make the material moldable, for example injectable into a space between the above-mentioned first mold part and second mold part. In such embodiments, subsequently molding 12 whichever of the substrate 102 and the support member 106 has not already been molded on the initially molded component may be implemented prior to the initially molded component returning to room temperature, and preferably while the initially molded component is at least 80°C. This may assist adhesion of the substrate 102 and the support member 106 to each other. In embodiments, in which the subsequent molding 12 comprises curing the precursor material, such maintenance of elevated temperature can also assist such curing.

[0170] In some preferred embodiments, the substrate 102 is initially molded 16 from a thermoplastic material, such as polycarbonate, poly ether ether ketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and / or polyethersulfone (PES), with a glass transition temperature of at least 140°C, and the support member 106 is subsequently molded 12; 12 A, 12B onto the substrate 102 via curing 12B the precursor material, e.g., siloxane precursor material.

[0171] In such embodiments, the mold may be opened 18A on one side while the initially molded substrate 102 remains in the mold. The third mold part may then close the mold to adopt the second mold configuration, and the precursor material, e.g., siloxane precursor material may be injected thereinto.

[0172] It is noted that the mold in the second configuration may be maintained at a temperature of 80 to 120°C and the precursor material, e.g., siloxane precursor material, may be injected while the mold is at this temperature.

[0173] Once filled with the precursor material, e.g., siloxane precursor material, the mold may be heated at 160 to 200°C until the precursor material is cured, the support member 106 is formed, and the support member 106 is adhered to the substrate 102, e.g. the thermoplastic of the substrate 102.

[0174] In more general terms, the elastomeric material may be molded at least partially against a surface of the substrate 102, e.g. the thermoplastic substrate 102, while in the mold.

[0175] The mold may then be cooled down, e.g., to between 40 and 80°C, and the two- component product may be released from the mold at this temperature. This may mitigate the risk of the two-component product sticking to the mold, as previously described.

[0176] The molding method disclosed herein has been described with reference to devices of Figs 1A to D and Figs 2A and 2B. The method is however not limited to manufacture of such devices but can be used for many other devices for many biological testing purposes. Some examples will be described below.

[0177] 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 may be arranged in a plurality of rows 105. In some embodiments, such as shown in FIG. 4, twelve rows 105 of chamber units 104 are defined in the substrate 102, with each row comprising eight chamber units 104, similarly to the row of chamber units 104 shown in FIGs. 1A and IB.

[0178] Thus, device 100 may have a ninety six chamber unit 104, e.g. well, configuration. In other embodiments, device 100 has a twenty four chamber unit 104, e.g. well, configuration. The number of chamber units 104 defined in the substrate 102, together with the shape and size of the substrate 102, e.g., and the device 100 as a whole, may correspond with number of wells, size and shape of well plates that are well-known and generally used in the field.

[0179] It is also reiterated that in some embodiments, such as 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 with the support member 106 comprising at least one structural feature 112, e.g. flexible protrusions, for contacting the biological material received in at least some, e.g. each, of the chamber units 104.

[0180] FIGs. 5A to 5C provide various views of a biological material testing device 100 according to an example which has a similar substrate 102 design to that shown in FIG. 4, but with the support member 106 having, for each chamber unit 104, the end portion 110 described above in relation to FIG. 2A. The molding 12 enables attachment of the substrate 102 and the support member 106 to each other, as previously described. The substrate 102 and the support member 106 are nonetheless shown separately from each other for clarity in the exploded perspective view provided in FIG. 5A, with the substrate 102 and the support member 106 being shown attached to each other in the transparent view of the device 100 provided in FIG. 5B and in the plan view provided in FIG. 5C.

[0181] In some embodiments, such as shown in FIG. 6, the biological material testing device 100 comprises a base member 132, such as a glass base member 132, on which the support member 106 and the substrate 102 are arranged.

[0182] The base member 132 may assist to reinforce the device 100. Alternatively, or additionally, the base member 132, e.g. the glass base member 132, may be optically transparent to enable analysis of the biological material through the glass base member 132 via optical microscopy.

[0183] The base member 132, e.g., glass base member 132, may, for some embodiments, also provide a smooth surface on which to mold the support member 106. Such a smooth surface may, in turn, assist the exterior surface of the support member 106 to have a smooth surface. This may facilitate analysis of the biological material through support member 106 via optical microscopy.

[0184] It is noted that in other embodiments in which the base member 132 is not included in the device 100, or more generally the exterior surface of the support member 106 is in contact with a surface of the mold during manufacture, the surface of the mold is preferably a polished surface.

[0185] Such a polished surface may assist to provide a relatively smooth exterior surface of the support member 106, which can ultimately facilitate analysis of the biological material through the support member 106 via optical microscopy.

[0186] It is noted at this point that the focus distance of many optical microscopy set-ups, e.g., employed for imaging stainings, may only be about 0.6 mm. The distance from the bottom of the base member 132, e.g., glass base member 132, and the support member 106 may accordingly be not more than 0.6 mm. However, other thicknesses may be used for different optical setups. More details of such optical setups and bottom plate material and design options are disclosed in the currently undisclosed pending patent application EP 23181319.7 and PCT / EP2024 / 066936 as well as in EP 23182831.0 and PCT / EP2024 / 067028 all of which are incorporated by reference herein in their entirety, but other devices are also possible.

[0187] The thickness of the optically transparent base member 132, e.g., glass base member 132, may be 0.10 to 0.30 mm, for example about 0.20 mm.

[0188] Such a thickness may account for the thickness of the support member 106, e.g., including the height H of flexible protrusions included in the support member 106.

[0189] More generally, the method 10 may include arranging at least part of a base member 132 in or adjacent to the mold, initially molding 16 the substrate 102 or the support member 106 on the base member 132. Following molding 12 of whichever of the substrate 102 and the support member 106 has not already been molded on the initially molded component, the base member 132 with the substrate 102 and the support member 106 molded thereon may be removed, e.g., released, from the mold.

[0190] In some embodiments, such as shown in FIGs. 7 and 8, at least one dual chamber unit 104 is defined in the substrate 102, with each of the dual chamber unit(s) 104 comprising a first chamber 134 and a second chamber 136 fluidly connected to each other via a bridge 138 (which may be referred to as a top fluid channel) defined between the first chamber 134 and the second chamber 136. Furthermore, the dual chamber units 104 can be interconnected by fluid channels in series, in parallel or in mixed configuration in one device. For example, in Fig. 8 multiple dual chamber units of each row 105 are interconnected in parallel between the first channel 137 and a second channel 139. The first channel may be referred to as a fluid supply channel and the second channel may be referred to as a fluid drain channel. However, as mentioned, other configurations may be defined. In the devices of Figs 7 and 8 the fluid channels 137, 138 and 139 are part of the substrate 102. They may be referred to as top-channels of the device.

[0191] Such a dual chamber unit 104 may have utility in studying metastasis of cancerous cells. It may be possible to assess cells that are released or emitted by any cell cultures that are provided in the first chamber 134. The first chamber 134 can therefore be used to perform a treatment on the cell culture(s), such as the application of a certain therapeutic fluid or drug, for example a chemotherapy treatment fluid, with the second chamber 136 capturing any released cells to analyze the effect of the treatment performed in the first chamber 134 on metastasis.

[0192] In such embodiments, the support member 106 may comprise, e.g., be defined by, the above-mentioned porous support member 106 for supporting the biological material, e.g., cells, received in the dual chamber unit 104. The support member 106 may, for example, include 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.

[0193] Any suitable number of dual chamber units 104 can be defined in substrate 102, for example two dual chamber units 104 in the embodiment shown in FIG. 7 and twenty-four dual chamber units 104 in the embodiment shown in FIG. 8. 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 incorporated herein by reference in their entirety.

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

[0195] More generally, method 10 can be used to form any device for testing biological material that has 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 in EP 23182831.0 and PCT / EP2024 / 067028 all of which are incorporated by reference herein in their entirety, but other devices are also possible.

[0196] Figs 9 A to 9 F are used to provide an example of how to apply the method 10 to make devices having one or more top channels and / or one or more bottom channels, as for example used in the device of Fig. 8 which have top channels 137, 138 and 139, as well as other features in the device. The Fig. 9A to F also serves to exemplify a mold system having the different parts to be used in the method. While the drawings of the example of Figs 9A to 9G show substrate and support member having two chamber units, the drawings may be considered to also represent cases where there are more of such chamber units such as for example in the devices of Figs 5 A to 5C, 6 and 8.

[0197] Figs 9A to 9F exemplify a method to manufacture a three component multi-chamber device 900 having parallel connected chamber units much like the device of Fig. 8, the differences being that the device 900 will only have one chamber (or container) per chamber unit as compared to the chamber units of Figs 7 and 8.

[0198] Fig. 9E shows cross sectional views of the device 900 to be made and Figs 9F and 9G show respective bottom and top views of the device 900. Fig. 9E shows three cross-sectional views of device 900. The lower two views IX and X show cross sections perpendicular to the cross section of the upper view along 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 walls has been indicated in dashed line format in Fig. 9F as one cannot see the chamber in this view through the support member 906 from the bottom side of the device 900.

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

[0201] On the bottom side of device 900 there is a support member 902 adhered to the substrate 902 according to principles as disclosed herein. The support member 906 comprises bottom channel 935 connecting the sub-chambers 975 situated below the membranes 942. Within the bottom view of Fig. 9F, channel 935 can be seen extending horizontally.

[0202] The support member 902 comprises end-portions 910 each of which is situated within one of the chamber units 934. Each endo-portion includes a membrane 942 with a circular shape with diameter 972 and a membrane thickness measured perpendicular to the plane it extends in. The circular shape can be seen from above or below in Figs 9F and 9G. Each end-portion also comprise a flange 966 each of which extends upwards from the bottom side of the substrate 902 to within a chamber unit 934 with some height smaller than the height of the chamber unit 934. Each one of the membranes 942 extends from a flange 966, or in other words is carried by or integrated with that flange.

[0203] Preferred membrane thickness is in the range of 5 to 100 pm, but other thickness can be used. This can be chosen based on the desired purpose of the device.

[0204] In some embodiments the membrane comprises or consists of a perforated membrane that comprises a plurality of pores. Each pore is preferably wide enough to allow passage of fluid possibly containing nutrients, drugs or other (bio)molecules such as proteins etc.

[0205] In some applications the pores are wide enough to at least partly allow passage of cells or particular types of cells.

[0206] In some embodiments the pore is narrow enough to prevent passage of cells. For example, the pore diameter may be in a range of 10 pm or less, for example it may be 5 pm. But other values as described hereinafter may be used. Therewith, the porous membranes can be used to support tissue samples such as cells while the fluid with all its contents indicated above can be provided to the cells by passing through the membrane using one or more of the channels in the device. Exemplifying membranes and channels are in the devices of Figs. 7 and 8 and those described in for example the EP 23181319.7 and PCT / EP2024 / 066936.

[0207] The support member 902 in its end-portions 934 can have other structural features 912. Thus, for example there may be a plurality of wells defined in a membrane. The bottom of such wells may again form a membrane or perforated membrane as described herein. Detailed examples of such well containing membranes have been described in the EP 23181319.7 and PCT / EP2024 / 066936.

[0208] The pores may for example be made using laser perforation as disclosed herein.

[0209] Figs. 9A to 9D each show three cross sectional views of a mold system as defined herein that is used in a method as defined herein to manufacture the device 900. It will be understood that the mold system can be used to manufacture other devices by adjusting its structural design. The mold system comprises a first mold part 950 and a second mold part 952 that can be combined to form a first configuration for an initial molding step 16 as described herein before. The first and second mold parts have a shape and size designed to mold the substrate 902 of the device 900. The lower two views I and II show cross sections perpendicular to the cross section of the upper view along directions indicated with I and II therein. 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) that together, during and after molding, define the shape and design of the substrate 902. In this case the first and second mold parts have protrusions 953 that at least partly define chamber units 934 as well as protrusions 955 that ultimately define top channels (much like the channels 137 and 139 of the device of Fig. 8) in substrate 902 that will be defined by filling of open spaces 954. In more detail, the protrusions 953 are in this case circular cylindrical with a diameter 170 and their cylinder axis vertical and in the plane of the cross- sectional drawing. They will eventually, after molding, define cylindrical chamber units 934 with a diameter 170 in substrate 902. Protrusions 955 are rectangular and extend along the first mold perpendicular to the plane of drawing of the cross-sectional view I. The first mold part 950 shows protrusion 957 abutting protrusion 953. Protrusions 957 are rectangular and extend only partly along the cylindrically shaped protrusion 953 such that eventually they will form the top channels that connect the chambers 934 to the top channels. The second mold part 952 has surface mold surface portions 980 (sometimes referred to as second mold surface) that will define substrate surfaces 982 that in a later stage of the process will define parts of the substrate onto which the support member 906 will be molded. These surfaces 982 will therefore be contact or adhesion surfaces as discussed herein before. The mold surface portions 980 may be geometrically designed to improve or add the adhesion of the support member to the substrate. In the Figs 9A to 9G, no particular geometric designs have been shown for clarity, but the first mold part surfaces 980 may have for example a specific surface roughness and or a corrugation of some type-

[0210] The first and second mold part are shown separate from each other in open configuration in Fig. 9A, and they are releasable attachable (separable) to each other in a leak tight fashion such that in the first mold configuration (when they are combined and held together), as shown in Figs 9B and 9D they can be used for molding by injection of 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 the liquid substrate material into the mold when it is in the closed molding configuration.

[0211] In an initial molding step 16 as described with reference to method 10 and Fig. 3, the first and second mold parts 950 and 952 are brought in the first configuration and under conditions described a substrate material will be molded into substrate 902 by e.g. injection molding the material into the mold openings 954 via injection opening 156. The result is shown in the cross-sectional views of Fig. 9B. As an example, the material injected may be one chosen from the group of thermoplastic materials such as a polycarbonate, as described herein before.

[0212] In a further step 18 A, the second mold part 952 is released from the first mold part 950 and the molded substrate 902 while leaving the substrate 902 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 and 953 now result in the ensemble of first mold part and substrate 902 having open spaces 964 with a diameter 170 and each forming an exposed part 964 of a substrate chamber 134. These open spaces will ultimately be occupied by the end portions 910 of the support material once molded. Furthermore, the substrate

[0213] In step 18 B the second mold part 952 is replaced with a third mold part 962 designed to fit to the first mold part 950 to form a second mold configuration in which a further open space (sometimes referred to as one or more second open spaces), remains in which the support member 906 will be molded. The design of the first mold part 950, the second mold part 960 and the molded substrate 902 now define the further open space. This second mold configuration is shown in Fig. 9D.

[0214] The third mold part 962 includes protrusions 961 having cylindrical shape with a diameter 172 and a cylinder axis oriented vertical and in the plane of 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 are such that they do not contact the protrusions 953 of mold part 950 in the second closed configuration. Therewith an open space remains in the form of an exposed part 964 in each of the chamber units 934 that may be occupied with the material that will form the end-portion part of the substrate member 906. In this case the further opens pace is thus designed to result in a support member 906 having a thin membrane 942 attached to flanges 966, where the thin membrane is between the protrusions 953 and 961. However, other designs may be used and the design of the protrusions 953 and 961 may be used to define end-portions as desired such as for example those defined herein. In this context the surface 964 may be referred to as a first mold surface and the surface 986 of protrusion 961 may be referred to as a third mold surface.

[0215] The third mold part also has one or more protrusions (only one shown for clarity) 963 that have a rectangular shape and extend perpendicular to the drawing of cross-sectional view VII. These protrusions will eventually form channels 935 in support member 906 that interconnect sub-chambers 975 beneath membranes 942.

[0216] In step 12, the support member 906 may be molded as described with reference to method 10 and Fig. 3 herein when the second mold configuration has been adopted. The material for molding is injected through opening 960 to form a result as shown in Fig. 9D. As the support member material is molded against part of the surface of the substrate, the contact between contact surfaces is intimate and the support member is molded largely conformal to the shape and irregularities in the surface of the substrate 902. This results in intimate contact with improved adherence and leakage properties as described hereinbefore.

[0217] After cooling down to the appropriate temperature as indicated herein before for method 10, the device in the form of the support member 906 molded onto the substrate 902 may be released from the first and second mold parts. The result will be device 900 of Figs 9E to 9F as described hereinabove. The device may be used as such if no further parts are needed. This may 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 fluid. The devices having the pillars as described herein may provide such examples. However, the device of Fig. 9E does have the bottom channels and these may need closure from the bottom side.

[0218] The mold system used of the method described with reference to the method of Figs 9A to 9D comprises the first mold part, the second mold part and the third mold part as defined herein.

[0219] In an alternative method according to the invention the molding of the substrate and the molding of the support member is not combined. Thus, the substrate is an already existing substrate, which may have been molded still, but is provided form stock. The first mold part is then designed as defined herein and to accommodate the substrate as provided from stock. The first and second mold parts are designed and used as described with reference to Figs. 9A to 9D. In such alternative method, a mold system according to the disclosure then only comprises the first mold part and the third mold part as defined herein.

[0220] To close the bottom channels off, the device may be adhered to base member 932 made of e.g. glass, or other suitable material such as for example those described herein. Fig. 9H shows the device of Fig. 9E to 9G with a base member 932 which closes off the channels 935 and the openings 975 on the bottom side of the device so that they are suitable for containing fluid. The channels are now suitable to provide fluid to the membranes 942 form the bottom side or drain fluid which has passed through such membrane from the top to the bottom side.

[0221] In an alternative method, the base member is already present in the third mold part 962, e.g. as an inlay or as pre-molded in third mold part 962, before injecting the mold material of the substrate member 906. In that case the support member may be molded not only conformal to the substrate 902, but also conformal to the base member 932.

[0222] A top cover plate can be added as desired. This may be releasably attachable to substrate 902. The top cover plate may be glass or polymer material. It may be the same material as that of the substrate material. The top cover is not shown in Fig. 9H for clarity.

[0223] It will be clear that devices as described herein of the two- or multi-component type can have designs as desired that differ from the examples provided herein. Thus, by adjusting the design and dimensions of protrusions 935 and 961 size and shape of chamber units 934 can be defined. Furthermore, their design and dimensioning can also be used to define open space between the protrusions 935 and 961 to therewith define the shape and dimensions of the end-portions 91o and the membranes 942. For example, the thickness of the membrane may be set with these design parameters. The membrane may also have wells defined therein. More detailed designs of membranes with perforations and / or wells including their desired dimensions have been described in the references to other documents provided herein. Also, the device 900 has only one chamber per chamber unit. There may be two or more chambers per chamber unit as for the Fig. 8 device 800. Fig. 9 shows device 900 with only two chambers and the corresponding channels. This is however only exemplary as the principles shown can be extended to the manufacture of devices having a different number of chambers and different number of channels etc. Thus, for example 96 chamber plates can be made with this method. More generally, the present disclosure proposes use of the device 100 according to any of the embodiments described herein for testing biological material, for example for testing cell groups, double cell layers, spheroids, organoids or biopsies.

[0224] The disclosure further contemplates a drug testing method comprising providing biological material in the at least one chamber unit 104 of a device 100 according to any of the embodiments described herein, and exposing the biological material to the drug to be tested.

[0225] In some embodiments, the providing of biological material in the at least one chamber unit 104 comprises culturing cells in the at least one chamber unit 104.

[0226] Alternatively, or additionally, the drug testing method may comprise monitoring a response of the biological material, e.g cultured cells, to the drug to be tested. This monitoring may be implemented in any suitable manner, for example by optical microscopy, e.g., through the support member 106 when the support member 106 is optically transparent.

[0227] Preferred combinations of silicone and (stiff) thermoplastics are silicone / polycarbonate, with the glass transition temperature (Tg) of polycarbonate being >140C or silicone / PEEK and silicone / PEEK blends:

[0228] The following exemplary commercially available elastomeric / soft materials (which are transparent or translucent and are food contact approved) can be used, in certain embodiments, as the 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® (F 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-12fl50S | 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-12340 NAT | TPE; Medalist® MD-12342 | TPE; Medalist® MD-12344 | TPE; Medalist® MD-12350 | TPE;

[0229] 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;

[0230] 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 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;

[0231] 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;

[0232] 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

[0233] 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;

[0234] 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;

[0235] 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;

[0236] Dynaflex™ G6713C | TPE; Dynalloy™ GP 7810-60T | TPE; Dynalloy™ GP 7810-70T | TPE;

[0237] Dynalloy™ OBC8200-BT50 | TPE; Estane® 58123 TPU | TPU-Polyether; Evoprene™ 019 | SBS;

[0238] 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-38052 XRD1 | 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;

[0239] Monprene® RG-10170 | TPE; Monprene® RG-15130 | TPE; Monprene® RG-15140 | TPE; Monprene® RG-15150 | TPE; Monprene® RG-15160 | TPE; Monprene® RG-15170 | TPE;

[0240] Monprene® RG- 18240 | TPE; Monprene® RG- 18250 | TPE; Monprene® RG- 18260 | TPE; Monprene® RG-18270 | TPE; Monprene® RG-19221 NAT | 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;

[0241] 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; THERMOLAST® 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;

[0242] 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.

[0243] Variations to the disclosed embodiments can be understood and effected by those skilled in the art in practicing the claimed invention, from a study of the drawings, the disclosure and the appended claims. In the claims, the word "comprising" does not exclude other elements or steps, and the indefinite article "a" or "an" does not exclude a plurality.

[0244] The mere fact that certain measures are recited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage. If the term "adapted to" is used in the claims or description, it is noted the term "adapted to" is intended to be equivalent to the term "configured to".

[0245] Any reference signs in the claims should not be construed as limiting the scope.

Claims

CLAIMS:

1. A method (10) for manufacturing a biological material 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 biological material received in the at least one chamber unit, the method comprising molding (12), using a mold, one of the substrate and the support member on at least part of the other of the substrate and the support member, which at least part of the other of the substrate and the support member is arranged in or adjacent to the mold during said molding.

2. The method (10) according to claim 1, comprising initially molding (16) the substrate (102) or the support member (106) to form an initially molded component, and subsequently molding whichever of the substrate and the support member has not already been molded on the initially molded component.

3. The method (10) according to claim 2, wherein initially molding (16) the substrate (102) or the support member (106) comprises heating a material above room temperature to make the material moldable to enable forming of the initially molded component, and wherein subsequently molding whichever of the substrate and the support member has not already been molded on the initially molded component is implemented prior to the initially molded component returning to room temperature, and preferably while a temperature of the initially molded component is at least 80°C.

4. The method (10) according to claim 2 or claim 3, comprising arranging at least part of a base member (132) in or adjacent to the mold, initially molding the substrate (102) or the support member (106) on the base member, and removing the base member with the substrate and the support member molded thereon from the mold.

5. The method (10) according to any one of claims 1 to 4, wherein the molding (12) of said 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 said curing comprises heating the precursor material.

6. The method (10) according to any one of claims 1 to 5, comprising releasing the substrate (102) together with the support member (106) from the mold while temperatures of the substrate and the support member are higher than room temperature, and preferably at least 40°C.

7. The method (10) according to any one of claims 1 to 6, comprising injection molding (12) said one of the substrate (102) and the support member (106) on said 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 mold from a first configuration used for molding said one of the substrate (102) and the support member (106) to a second configuration used for molding said 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 softer and / or more flexible material than that forming the substrate (102).

10. The method (10) according to any one of claims 1 to 9, wherein the support member (106) is formed from a more biocompatible material than that forming the substrate (102) so that cells and / or tissue preferentially adhere(s) 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 that is bulk-modified with moieties that each include a polar group, polar groups of said moieties being available at a surface of the support member arranged to contact the biological material received in the at least one chamber unit (104); optionally wherein the moieties comprise fatty acid moieties, the polar groups comprising carboxylic acid groups of said fatty acid moieties.

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

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

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 received in the at least one chamber unit (104); optionally wherein the at least one structural feature comprises, per chamber unit, 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 unit (104) comprises a plurality of chamber units; optionally wherein the chamber units are arranged in one or more rows (105).

17. A mold system for molding a biological testing device using a method as disclosed herein, the mold system comprising: 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 configured to be combined to form a second mold configuration arranged to hold one of the substrate and support member such that in the second mold configuration the first mold part, the second mold part and the one of the substrate and support member define one or more openings in which the other of the substrate and support member can be molded onto the one of the substrate and support member.

18. A biological material testing device (100) comprising: a substrate (102) in which at least one chamber unit (104) is defined; and a support member (106) for supporting biological material received in the at least one chamber unit, one of the substrate and the support member being molded on the other of the substrate and the support member.

19. Use of the device (100) according to claim 18 for testing biological material.

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

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