Biomaterial testing device and method for manufacturing and using the biomaterial testing device

Abstract

A biomaterial testing device 100 is provided, comprising a substrate 102 on which at least one chamber unit 104 is defined, and a support member 106 for supporting a biomaterial received in at least one chamber unit. One of the substrate and the support member is molded on the other of the substrate and the support member. A method 10 for manufacturing such a biomaterial testing device is also provided. Furthermore, a method and use of the device for testing a biomaterial is provided, for example, in drug testing where the biomaterial is exposed to a drug to be tested.

Description

Technical Field

[0007]

[0001] The present invention relates to a method for manufacturing a biological material inspection device.

[0002] The present invention also relates to the biological material inspection device itself obtainable by this method and a biological inspection device.

[0003] The present invention also relates to a mold used in this manufacturing method.

[0004] The present invention further relates to the use of a device for inspecting such biological materials.

[0005] The present invention also further relates to a drug inspection method in which this device is used.

Background Art

[0006] In vitro inspection of mammalian cells or tissues is an important technique for obtaining clinically important information on mammalian materials under study. For example, such inspections are performed on biopsied mammalian cell or tissue materials to determine abnormalities or diseases in such mammalian materials, or to expose diseased mammalian materials to drugs such as experimental drugs and monitor the response of the diseased mammalian materials to such exposure. This approach is frequently used, for example, in oncological treatments. From this, important insights can be obtained regarding a method for effectively treating an individual's disease without exposing the individual to various drugs that may be effective but undesirable for several reasons, including drug toxicity.

[0007] Drugs are sometimes tested for cardiotoxicity. Many drugs can adversely affect myocardial contractility and profile. Drug-induced cardiotoxicity is a major side effect seen in some clinically important drugs. This toxicity has led to the post-market withdrawal of numerous pharmacologically active drugs and has also limited the efficacy of other clinically useful drugs. Cardiovascular safety concerns have resulted in the recall of nearly 10% of drugs from the global market over the past 40 years (see, for example, Chapter 1 of "Cardiotoxicity" edited by Wenyong Tan, and "Drug Induced Cardiotoxicity: Mechanism, Prevention and Management" by Mina T. Kelleni and Mahrous Abdelbasset, 2018).

[0008] Thus, drug-induced cardiotoxicity is a major reason for rejecting compounds in preclinical and clinical development, reflecting the severity of cardiotoxicity as an adverse effect observed during new drug development. Assessment of the risk of drug-induced cardiotoxicity, including QT interval prolongation, is now considered an essential part of standard preclinical evaluation of novel chemical components (see "Drug Induced Cardiotoxicity: Mechanism, Prevention and Management" for further details).

[0009] Cardiotoxicity can be tested using animal models, such as rats. However, these models have drawbacks and cannot always reliably predict the effects that humans will experience. This can lead to the discarding of potentially beneficial drugs, but it can also mean that potentially harmful drugs will proceed to human trials.

[0010] Another method for evaluating cardiotoxicity is to test the effects of a drug in vitro. The drug to be tested is added to a collection of cardiomyocytes grown in vitro. The effects of electrical stimulation on the collection of cardiomyocytes are then examined. An example of a state-of-the-art tool is "HeartDyno" (trademark), developed by James Hudson's group (see, for example, Mills et al., "Functional screening in human cardiac organoids reveals a metabolic mechanism for cardiomyocyte cell cycle arrest," Proceedings of the National Academy of Sciences, 2017, Vol. 114, No. 40).

[0011] This type of tool is included in this specification under the term “support member” and comprises an egg-shaped well having two small protrusions, e.g., pillars, at the bottom. Such support members can be manufactured using thin-film technology. For example, a mold can be fabricated on a wafer using SU-8 photolithography, resulting in features with a depth of 700 μm. Polydimethylsiloxane (PDMS) can then be poured onto these features and cured. After removing the PDMS from the wafer, a sample with a diameter of 6 mm can be obtained by punching.

[0012] These samples are placed, for example, in the wells of a 96-well substrate and glued to the bottom of the substrate using a silicone adhesive / glue. A mixture of cardiomyocytes, cardiac fibroblasts, collagen, DMEM, NaOH, and Matrigel can then be added to the wells and supported on the samples. It should be noted that one or more wells defined in such a substrate will be referred to herein using the more general term “chamber unit.” The cardiomyocyte tissue forms over several days. This tissue exhibits spontaneous contraction, but also contracts during electrical stimulation. During contraction, two supports deflect, and this deflection is analyzed using a video analysis algorithm.

[0013] Fluid devices are also known that include two or more fluid channels for transporting fluid to one or more chamber units defined on a substrate of the fluid device, such as a culture chamber. One channel can be used, for example, to supply nutrients and oxygen to each of the chamber units and to remove metabolites such as carbon dioxide from there. Further channels can be used to supply drugs to the chamber units. Such devices include support members, such as porous support members, for supporting biomaterials such as cell populations, double cell layers, spheroids, organoids, and biopsy specimens received within the chamber units. [Overview of the Initiative] [Problems that the invention aims to solve]

[0014] For example, various technical challenges arise in adhering the above-described type of substrate to the above-described type of support member. It is difficult to perform such adhesion in a sterile manner and / or without unintentionally creating areas where fluid leakage may occur between the substrate and the support member. Furthermore, additional manufacturing processes are required to adhere the substrate and the support member to each other. [Means for solving the problem]

[0015] The present invention is defined by the claims.

[0016] According to an example in one aspect of the present disclosure, a method is provided for manufacturing a biomaterial testing device, the device comprising a substrate on which at least one chamber unit is defined, and a support member for supporting a biomaterial received in at least one chamber unit, the method comprising the step of molding one of the substrate and the support member onto at least a portion of the other of the substrate and the support member using a mold, wherein at least a portion of the other of the substrate and the support member is positioned in or adjacent to the mold during the molding step.

[0017] The method involves shaping one of the substrate and support member onto at least a portion of the other, thereby shaping the former and bonding it to the latter in the same step. In this way, a separate assembly step of bonding the substrate and support member to each other, such as by gluing and / or clamping, can be omitted. No adhesive / glue is required to keep the substrate and support member bonded to each other. Thus, the method provides a relatively rapid and reliable process for manufacturing a device in which the substrate and support member can be considered as a single integrated piece.

[0018] Furthermore, support members, such as silicone support members, are secured by their shape within a frame defined by the substrate, such as a polycarbonate frame. For example, the surfaces of the substrate and support members that will come into contact within the device have a geometric structure or shape to promote adhesion. Such structures or shapes are selected from the group consisting of rough surfaces, corrugations, protrusions, and recesses. However, other structures may be used additionally or as substitutes.

[0019] In at least some embodiments, the molding step includes injection molding one of the substrate and support member onto at least a portion of the other of the substrate and support member. Injection molding provides a cost-effective and scalable method for manufacturing devices.

[0020] In some embodiments, the method comprises the steps of initial molding a substrate or support member to form an initial molded component, and subsequently molding the unmolded of the substrate and support member onto the initial molded component. In such embodiments, the method includes two-stage injection molding, for example, so-called "2K" injection molding.

[0021] Alternatively, or in addition, the step of molding one of the substrate and support member onto at least a portion of the other of the substrate and support member includes curing a precursor material. The precursor is preferably in a liquid state when added to the mold. In the case of injection molding, molding includes 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 convert the precursor material into a solid support member material. A precursor material that undergoes a chemical transformation is preferred. The use of a precursor material is advantageous for molding substrates and / or support members containing rubber materials. Such materials are difficult to shape after formation. For example, a support member disclosed herein containing a rubber-type silicone polymer is preferably made using a liquid precursor material that solidifies into a silicone polymer rubber by a chemical reaction of the precursor components. Other rubber materials can also be used, and these other rubber materials can benefit from the use of precursor materials in this method.

[0022] Curing involves heating the precursor material to a temperature higher than, for example, 160°C, e.g., 160–200°C. Heating at such temperatures contributes to the curing of the precursor material and ensures adhesion of the substrate and the other support member, e.g., at least a portion of the initial molded component. Heating is used to induce chemical and / or physical transformations.

[0023] In some embodiments, the step of initial molding a substrate or support member includes heating the material to a temperature above room temperature, often 200-300°C in the case of thermoplastic materials, commonly known as thermoplastics, to enable the formation of the initial molded components. It should be noted that the term “thermoplastic material” as used herein is intended to refer to polymers that can be softened by heating and subsequently processed, in particular, molded.

[0024] In such an embodiment, the step of subsequently molding the as-yet-unmolded one of the substrate and the support member onto the initial molding component is preferably carried out before the initial molding component returns to room temperature, and more preferably while the temperature of the initial molding component is at least 80 °C. This contributes to adhering the substrate and the support member to each other. In embodiments where the subsequent molding step includes curing the precursor material, maintaining such a high temperature can also contribute to such curing.

[0025] In some embodiments, the method comprises the step of disposing at least a portion of the base member within or adjacent to the mold, and the step of initially molding the substrate or the support member onto the base member.

[0026] Following the step of molding the as-yet-unmolded one of the substrate and the support member onto the initial molding component, the base member with the substrate and the support member molded thereon is removed from the mold, for example demolded.

[0027] The base member contributes to reinforcing the device. Alternatively or in addition, the base member, such as a glass base member, is optically transparent to enable analysis of biological material through the glass base member by an optical microscope.

[0028] The base member, such as a glass base member, also provides a smooth surface onto which the support member will be molded. Such a smooth surface consequently contributes to the outer surface of the support member having a smooth surface. This facilitates analysis of biological material through the support member by an optical microscope.

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

[0030] Such polished surfaces contribute to providing a relatively smooth outer surface of the support member, which ultimately facilitates the analysis of biomaterials through the support member using an optical microscope.

[0031] In some embodiments, the method includes the step of releasing the substrate together with the support members from the mold while the temperature of the substrate and support members is above room temperature, preferably at least 40°C. This minimizes the risk of one or both of the substrate and support members becoming stuck to the mold.

[0032] In some embodiments, the method includes the step of adjusting the mold from a first configuration used to mold one of the base material and support member to a second configuration used to mold the other of the base material and support member. In such embodiments, the step of adjusting the mold from the first configuration to the second configuration includes switching or exchanging the second mold part for a third mold part while the initially molded components remain received within the first mold part between the first and second mold parts. Such switching includes, for example, displacing the second mold part to open the mold and then using the third mold part to close the mold again.

[0033] In preferred embodiments, the method preferably comprises a mold including a second mold configuration for forming a support member on a substrate, the second mold configuration comprising a second mold component and a third mold component separable from a first mold component, together forming the second mold configuration. In some embodiments, the substrate is pre-formed and inserted into the first mold component before the support member is formed. In other embodiments, the method comprises a mold including a first mold configuration for forming a substrate, the first mold configuration comprising a first mold component and a second mold component, the second mold component being separable from the first mold component and from the substrate once formed, such that the substrate remains inside the first mold component. In such embodiments, the step of adjusting the mold from a first configuration to a second configuration thus includes switching or replacing the second mold component with a third mold component while the substrate initially formed between the first and second mold components remains received inside the first mold component. Such switching may include, for example, displacing a second mold part to open the mold, and then using a third mold part to close the mold again.

[0034] In preferred embodiments, which also use a second mold configuration and optionally, but preferably, a first mold configuration, the support member is molded on a substrate. Preferably, the substrate contains or consists of a thermoplastic or thermosetting material. Preferably, such material is transparent. For example, preferably the material contains at least one component selected from the group consisting of polycarbonate, polyetheretherketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and polyethersulfone (PES). The most preferred material is polycarbonate. Such materials generally have one or more of the following desired properties, namely, thermal properties as further described herein, transparency to visible light, rigidity (e.g., not rubber), and availability / cost. A substrate containing or consisting of any one of these materials is preferably combined with a support member containing a rubber material. Preferred rubber materials include silicone, for example, one or more silicone polymers as described herein. Preferably, the silicone rubber material is molded using a precursor material as described herein, which undergoes curing as described herein. Most preferably, the silicone rubber material is a silicone rubber material modified with a polar group such as a carboxylic acid group or a group of its conjugate bases.

[0035] In some embodiments, the support members are formed from a material that is more flexible than the material forming the substrate, for example, having a lower Shore A hardness and / or greater flexibility. Thus, the mechanical properties of the support members are suitable for supporting biomaterials cultured in the chamber unit, such as cells or tissues, while the substrate contributes more significantly to the structural rigidity of the device.

[0036] Alternatively, or in addition, the support member is formed from a material that is more biocompatible than the material forming the substrate, so that cells and / or tissues preferentially adhere to the support member. Thus, by using material selection, cell / tissue proliferation can be guided to the location where such cell / tissue proliferation is intended, for example, in the area of ​​a device that facilitates the observation and / or examination of cells / tissues.

[0037] In some embodiments, the support member comprises silicone, preferably the silicone comprises or consists of a silicone polymer in the form of rubber. Such a silicone support member is optically transparent and exhibits limited autofluorescence, so that various conventional optical inspection techniques and protocols, for example with or without staining, can be used in inspections performed using the device. Furthermore, the silicone has suitable biocompatibility, particularly when modified so that polar groups are available on the biomaterial contact surface of the silicone substrate.

[0038] In more general terms, the support member comprises a bulk-modified polymer material with sites containing polar groups, the polar groups of which are active on the surface of the support member positioned to contact a biomaterial received within at least one chamber unit. Such polar groups contribute to making the support member biocompatible, for example, by applying a protein such as fibronectin to the polar-functionalized surface of the support member.

[0039] 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 allows for the molding of a support member onto the substrate, and in this case, for example, such molding involves curing the precursor material at a relatively high temperature.

[0040] Alternatively, or in addition, the substrate may be formed from a thermoplastic, preferably one or more selected from polycarbonate, polyetheretherketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and polyethersulfone (PES).

[0041] In some embodiments, the substrate provides the sidewalls of at least one chamber unit, and within at least one chamber unit is a central portion of the support member, from which several rib elements extend, each engaging with the sidewalls of the corresponding chamber unit. Such rib elements contribute to minimizing deformation of the central portion while keeping the shear stress large enough to help maintain the support member in place.

[0042] In some embodiments, the support member comprises at least one structural feature that contacts the biomaterial received within at least one chamber unit. Such a structural feature provides a shape or profile suitable for growing cells or tissues on it. The shape or profile of the structural feature is selected, for example, depending on the type of cells or tissue to be grown and / or the examination to be performed on the biomaterial. The structural feature can be selected from a group consisting of membranes, porous membranes, multiple wells each having a membrane or porous membrane, and membranes having openings or slits for clamping biomaterials such as biopsy specimens. However, other structural features may be selected.

[0043] In some embodiments, at least one structural feature comprises, for each chamber unit, a pair of flexible projections, such as flexible posts or pillars, that support tissue on top of and between. In such embodiments, the device is used to examine muscle tissue, such as cardiac tissue, that has grown between and around the pair of projections. When the muscle tissue contracts, the pair of projections flex, and the resulting displacement of the projections can be measured to determine the force exerted by the tissue contraction.

[0044] In some embodiments, at least one chamber unit is composed of multiple chamber units. Therefore, the examination of biomaterials can be performed simultaneously in different chamber units. In such embodiments, the chamber units can be arranged in one or more rows, for example, in a manner similar to a conventional well plate.

[0045] In another embodiment, a molding system is provided for molding a biological testing device using a method as disclosed herein. The molding system comprises a first molding component and a third molding component separable from the first molding component, the first and third molding components being designed and arranged together to form a second molding configuration, which is configured to hold one of a substrate and a support member, in which the first molding component, the second molding component, and one of the substrate and support member surround one or more second open spaces in which the other of the substrate and support member can be molded on one of the substrate and support member. In a preferred embodiment, 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 an embodiment, the second molding configuration is configured to mold the support member on the substrate.

[0046] Preferably, the first molded component comprises at least one first molded surface defining the end chamber surface. The first molded surface can be used in this case to design the end surface which will ultimately be used to support a biomaterial. Preferably, the first molded component comprises or consists of a chamber projection including the first molded surface, the chamber projection extending into the mold space of the second molded configuration, and when the molded configuration holds the substrate, the projection occupies (e.g., fills) at least a portion of the chamber of the substrate such that the first molded surface is exposed in one or more second open spaces of the second molded configuration. The first molded surface can be shaped in this case to provide the end of the support member with a desired design as defined herein, such as a pillar, membrane, well, etc. The first molded surface can have any desired shape as defined herein, such as being flat, curved, projection, or recess or concave, depending on the desired shape of the support member to be formed on this surface.

[0047] Using these mold designs, a chamber unit can be fabricated in which parts of the wall are defined by a substrate, and parts of the wall are defined by support members, and at least a portion of the surface of the support members is exposed within the chamber to support biomaterials.

[0048] The first mold component is further positioned such that, when it holds the substrate, at least one, preferably more, contact surfaces of the substrate are exposed within one or more spaces of the second mold configuration. This allows the support member to be molded on these contact surfaces using the second mold configuration.

[0049] The first molded component may have one or more protrusions that occupy (e.g., fill) channels in the substrate.

[0050] Preferably, the third molded component comprises at least one third molded surface defining an end. The first molded surface in this case can be used to design an end surface which will be used to support a biomaterial in the finished device. The third molded surface is positioned to define one or more of a flange, membrane, or pillar structure. Preferably, the first and third molded surfaces are positioned within the corresponding molded components in the second mold configuration such that they define a portion of one or more second open spaces in which the end of the support member is defined. Preferably, the third molded surface is configured as a projection positioned on the third molded component so as to extend toward the first molded surface within one or more open second openings in the second mold configuration. Preferably, the projection defines a sub-chamber in the support member. Preferably, the third molded component comprises one or more channel projections for forming one or more channels in the support member. For example, some of these protrusions are arranged to form channels that connect to one or more sub-chambers.

[0051] Preferably, the mold system further comprises a second mold component, separable from the first mold component, which is designed and positioned to combine with the first mold component to form a first mold configuration, in which the first mold configuration, the first and second mold components enclose one or more first open spaces in which a substrate and / or a support member can be molded. The second mold component is not required in methods that use only the second mold configuration. For example, the second mold component is useful and advantageous when a substrate that has been pre-molded in the process is used in a method that includes a step of molding a substrate.

[0052] Preferably, the second molded component comprises a second molded surface that defines the contact surface of the substrate to be molded. This second molded surface can be shaped to improve the adhesion of the support member to the substrate. Preferably, the second contact surface comprises at least one of corrugations, depressions, protrusions, and surface roughness. Any shape that increases the contact surface area compared to a flat contact surface is beneficial for adhesion.

[0053] Preferably, the second mold component comprises further chamber projections arranged to extend into one or more first open spaces of the first mold configuration, such that in the first mold configuration, a chamber unit is defined on the substrate by the combination of the chamber projections of the first mold component and the further chamber projections.

[0054] The mold system is preferably an injection molding mold system. Preferably, one or more of the molded parts have one or more openings for injecting the material to be molded into the base material and support member. Preferably, the molded parts are made of a metal such as steel.

[0055] The shapes and sizes of the first molded part, the second molded part, and the third molded part are preferably such that the device, preferably the substrate of such device, includes at least one, preferably more chambers.

[0056] In another embodiment, a biomaterial inspection device is provided, comprising a substrate on which at least one chamber unit is defined, and a support member for supporting a biomaterial received in at least one chamber unit, wherein one of the substrate and the support member is molded on the other of the substrate and the support member.

[0057] The device can be obtained by any of the embodiments described herein. The device can be obtained by molding one of the substrate and the support member onto the other of the substrate and the support member.

[0058] The device preferably comprises at least one chamber, the at least one chamber defined by a portion of the wall of the chamber unit and a portion of the surface of a support member. The portion of the support member preferably comprises or consists of an end as defined herein. Preferably, the end is positioned within the chamber unit such that it forms the bottom of the chamber or at least a portion of the bottom of the chamber.

[0059] A chamber unit may have one chamber or multiple chambers. A device may have multiple chambers. These may be arranged as one or more arrays. In some embodiments, the device includes fluid channels between the chambers. Preferably, these channels are located in a substrate. In some embodiments, the chambers are arranged to connect multiple chambers. Alternatively or in addition, a support member includes at least one, for example, multiple, fluid channels. In some embodiments, the device includes at least one membrane, such membrane being part of the support member. Preferably, the membrane is part of an end. In some embodiments, such membrane is porous. In some embodiments, the end includes at least one well. In some embodiments, the well includes a membrane. The end may include a flange extending at least partially into the chamber unit. The flange is arranged to support or be integral with the membrane, or to be integral with the portion of the end that includes a well as defined herein. Preferably, the end includes a porous membrane and a flange, and the support member is further arranged to define sub-chambers bounded by the membrane and connected to one or more fluid channels located in the support member.

[0060] The embodiments described herein in relation to the method are applicable to the device, and the embodiments described herein in relation to the device are applicable to the method. Therefore, preferred materials described in relation to the method are also preferred materials for the device itself. Similarly, preferred structural device features of a device described in relation to the method disclosed and specified herein are also preferred features for the device itself.

[0061] In yet another embodiment, the use of a device according to any of the embodiments described herein for inspecting biomaterials is provided.

[0062] In a further embodiment, a drug testing method is provided, comprising the steps of: providing a biomaterial in at least one chamber unit of a device according to any embodiment described herein; and exposing the biomaterial to a drug to be tested.

[0063] In some embodiments, the step of providing biomaterial in at least one chamber unit includes culturing cells in at least one chamber unit.

[0064] Alternatively, or in addition, a drug testing method may include a step of monitoring the reaction of a biological material, such as cultured cells, to the drug to be tested. This monitoring step may be carried out in any suitable manner, for example, by an optical microscope, or through a support member, for example, if the support member is optically transparent.

[0065] These and other aspects of the present disclosure will become apparent from the embodiments described below and will be described with reference to them.

[0066] To better understand this disclosure and to more clearly illustrate how it is implemented, refer hereby to the attached drawings as merely examples. [Brief explanation of the drawing]

[0067] [Figure 1A] This diagram schematically illustrates a method for manufacturing a biomaterial testing device according to the first example. [Figure 1B] This diagram schematically illustrates a method for manufacturing a biomaterial testing device according to the first example. [Figure 2A] This figure shows some example support members that will be included in the device. [Figure 2B] This is a diagram of another exemplary support member that will be included in the device. [Figure 3] This is a flowchart illustrating a method for manufacturing a biomaterial testing device using another example. [Figure 4] This figure shows a device according to the second example. [Figure 5A] This diagram shows the device from the third example, presented from various perspectives. [Figure 5B] This diagram shows the device from the third example, presented from various perspectives. [Figure 5C] This diagram shows the device from the third example, presented from various perspectives. [Figure 6] This figure shows a device according to the fourth example. [Figure 7] This figure shows a device according to the fifth example. [Figure 8] This figure shows a device according to the sixth example. [Figure 9A] This figure shows a set of mold parts for forming a mold of a first mold configuration, which will be used to mold the substrate of the devices shown in Figures 9E-9H using the method disclosed herein. [Figure 9B] This figure shows a first mold configuration using the molded part shown in Figure 9A, in which the substrates of the devices shown in Figures 9E to 9H are molded inside. [Figure 9C] This figure shows the replacement of molded parts to provide a second mold configuration for molding the support members of the devices shown in Figures 9E to 9H according to the method disclosed herein. [Figure 9D]This figure shows the replacement of molded parts for providing a second mold configuration for molding the support members of the devices shown in Figures 9E to 9H according to the method disclosed herein. Figure 9D shows the second mold configuration and the support members molded therein. [Figure 9E] This is a cross-sectional view of a device formed according to the method disclosed herein. [Figure 9F] This is a top view of a device molded according to the method disclosed herein. [Figure 9G] This is a bottom view of a device molded according to the method disclosed herein. [Figure 9H] This is a cross-sectional view of a device formed according to the method disclosed herein. [Modes for carrying out the invention]

[0068] This disclosure will be explained with reference to these figures.

[0069] Detailed descriptions and specific examples illustrate exemplary embodiments of the apparatus, systems, and methods, but it should be understood that they are intended for illustrative purposes only and are not intended to limit the scope of this disclosure. These and other features, aspects, and advantages of the apparatus, systems, and methods of this disclosure will be better understood from the following description, the accompanying claims, and the accompanying drawings. The figures are schematic and not drawn to exact scale. It should also be understood that the same reference numerals are used throughout these figures to indicate the same or similar parts.

[0070] A biomaterial testing device is provided, comprising a substrate on which at least one chamber unit is defined, and a support member for supporting a biomaterial received within the at least one chamber unit. One of the substrate and the support member is molded on the other of the substrate and the support member. A method for manufacturing such a biomaterial testing device is also provided. Furthermore, uses and methods are provided for using the device to test a biomaterial, for example, in drug testing where the biomaterial is exposed to a drug to be tested.

[0071] Figures 1A and 1B schematically illustrate an example device design and manufacturing method 10 according to the present disclosure. Method 10 is for manufacturing a device 100 for examining biomaterials. For this purpose, the device 100 comprises a substrate 102 on which at least one chamber unit 104 is defined. The substrate 102 shown in Figures 1A and 1B has a plurality of such chamber units 104, in this case eight. The device 100 further comprises a support member 106 for supporting the biomaterial when the biomaterial is received in the chamber units 104, for example, in each of the chamber units 104 (the biomaterial is not visible in Figures 1A and 1B).

[0072] Examples of biomaterials include cell populations, double cell layers, spheroids, organoids, or biopsy specimens, but other materials may also be used.

[0073] Each chamber unit 104 extends between the top and bottom surfaces of the base material 102 and has a height in the range of 1.8 to 2.6 mm, for example, about 2.2 mm. Alternatively or in addition, the diameter of each chamber unit 104 is in the range of 3 to 20 mm, such as 3 to 10 mm, for example, about 6 mm.

[0074] Such dimensions strike a balance between the requirement to keep the chamber units 104 as small as possible in order to define, for example, as many chamber units 104 as possible within the substrate 102, and the requirement that the chamber units 104 provide sufficient space for the examination of biomaterials, for example, for cell capture and subsequent analysis.

[0075] The chamber unit 104 can have any suitable shape. For example, from a fluid dynamics standpoint, a generally cylindrical shape, such as those shown in Figures 1A and 1B, is preferred in order to reduce the risk of fluid accumulation in the corners of the chamber unit 104 and / or to ensure ease of use with existing analytical instruments. However, shapes other than cylindrical, such as those with a square or rectangular cross-section, can also be considered.

[0076] In some embodiments, such as those shown in Figures 1A and 1B, the base material 102 provides a side wall 108 of at least one chamber unit 104, for example, a cylindrical side wall 108, and the support member 106 provides an end 110 from which the side wall 108 extends. By combining the end 110 with the side wall 108 of each chamber unit 104, a container is obtained in which a biomaterial can be placed together with, for example, a cell culture medium. Generally, the end 110 can be called the bottom of the chamber, in which case the chamber is also called the container.

[0077] Generally, each chamber or container has a chamber depth measured along the side wall from the top surface to the edge of the substrate. In the examples in Figures 1A and 1B, the height is equal to the chamber depth, but this is not mandatory. In a variation of this example, as will be disclosed later herein, a support member may extend from the bottom surface of the chamber unit into the chamber unit, and the chamber depth may be less than the height by the distance the support member extends from the bottom surface into the chamber unit.

[0078] The thickness of the support member 106, for example, the end portion 110 provided by the support member 106, is selected to allow analysis of the biomaterial received in the chamber unit 104 through the support member 106.

[0079] Such analysis includes optical microscopy, with or without staining. For this purpose, the support member 106, for example, at least the end 110 provided by the support member 106, is optically transparent and has a thickness of, for example, up to 0.4 mm. Optical in this sense refers to a spectral range including one, more, or all of visible light, UV light, and near-infrared light.

[0080] In at least some embodiments, at least one chamber unit 104 is composed of multiple chamber units 104. In this case, multiple chamber units are present in the substrate and therefore in the device. Thus, the examination of biomaterials can be performed simultaneously in different chamber units 104. In such embodiments, the chamber units 104 can be arranged as one or more rows 105. Figures 1A and 1B show one such row 105 composed of eight chamber units 104, but as will be apparent from further examples described below herein, two or more rows 105 can also be conceived, for example, in which each row 105 is composed of eight, fewer than eight, or more than eight chamber units 104.

[0081] In some embodiments, such as those shown in Figures 1A and 1B, the device 100, for example its substrate 102, includes one or more mounting features 111, such as tabs, for enabling the device 100 to be mounted on and / or inside a holder (not shown). In such embodiments, the holder is configured, for example, to support multiple such devices 100 simultaneously on and / or inside it.

[0082] The holder, for example, holds 12 of the devices 100 shown in Figures 1A and 1B, and by combining the holder with the 12 devices 100, it defines 96 chamber units 104, for example, a 96-well plate.

[0083] In some embodiments, one or more of the devices 100 are individually removable from the holder.

[0084] This allows for a more flexible holder-device configuration. In a non-limiting example where 12 devices 100 can be mounted in the holder, if the experimenter does not wish to use all 96 chamber units 104 at once, one or more of the devices 100 can be removed.

[0085] Biomaterials tend to be required to remain alive for at least part of the examination performed using device 100. Therefore, device 100, for example, at least its support member 106, is formed from a suitable biocompatible material. In some embodiments, the support member 106 is formed from a biocompatible polymer material such as silicone, to which a suitable cell culture protein such as fibronectin may or may be applied. Suitable materials are mentioned herein.

[0086] In some embodiments, the support member 106 comprises a bulk-modified polymer material with portions each containing polar groups, the polar groups of the portions being effective on the surface of the support member 106 which is positioned to contact a biomaterial received within at least one chamber unit 104.

[0087] Such polar groups contribute to making the support member 106 biocompatible, for example, by applying proteins such as fibronectin to the surface of the support member that has been functionalized with polar groups.

[0088] In some embodiments, the moiety includes a fatty acid moiety, and the polar group includes a carboxylic acid group of the fatty acid moiety.

[0089] We will specifically mention that silicones are bulk modified with polar group-containing sites, such as fatty acids, via crosslinking, for example, between silicone-hydrogen bonds and C=C bonds in unsaturated fatty acid sites.

[0090] This crosslinking is catalyzed by a platinum catalyst.

[0091] In this regard, see, for example, the materials and methods described in International Applications 2021 / 058657 and 2019 / 015988. These applications further describe the processing of preferred materials to be used in the manufacture of the support members specified herein, and the entirety of these applications is incorporated herein by reference. Thus, referring to these documents, in some embodiments, 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. Phosphate groups and their conjugate bases are preferred over sulfonic acid groups, and carboxylic acid groups are preferred over phosphate groups. Residues supporting such groups are readily usable in bulk modification because, in such modification methods, their precursors having acidic groups in the form of conjugate bases with metal counterions and such residues mix well with the other components formed into the elastomer. The acidic groups are selected according to the required pKa (as already detailed herein).

[0092] In some preferred embodiments, the residue contains or consists of an aliphatic moiety comprising three or more carbon atoms and fewer than 50 carbon atoms, with one or more acidic groups covalently bonded to the aliphatic moiety, and the aliphatic moiety covalently bonded to the elastomer bulk. The aliphatic moiety may be linear or branched. The aliphatic moiety may contain one or more carbon-carbon double or triple bonds, or aryl or benzene units. The aliphatic moiety may contain cyclic units such as cyclohexyl or cyclopentyl. The aliphatic moiety is preferably a saturated hydrocarbon moiety. The aliphatic moiety preferably contains only carbon and hydrogen atoms.

[0093] In some embodiments, the aliphatic moiety is a linear chain bonded to the elastomer bulk at its termini. In some embodiments, the aliphatic moiety further does not contain carbon-carbon triple bonds to increase the flexibility of the residues.

[0094] In some embodiments, the residues within the bulk modified elastomer are distinct from each other, each selected as specified herein. In some embodiments, at least a portion of the residues are bonded to the elastomer bulk via two covalent bonds. This is due, for example, to the presence of a precursor having two ethylene bonds to participate in the modification reaction. In some embodiments, this portion contains at least five carbon atoms, more preferably at least ten carbon atoms. Preferably, the aliphatic portion contains fewer than 40 or fewer than 30 carbon atoms. A linear portion is preferred, but this is not essential. The number of carbon atoms in such a portion is preferably 5 to 30, more preferably 5 to 20 or 5 to 15. This 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.

[0095] In some embodiments, the residue is the remainder of an unsaturated fatty acid precursor covalently bonded to the elastomer bulk via the reaction of one or more of its ethylene groups (carbon-carbon double bonds), if present.

[0096] For example, an unsaturated fatty acid residue is one or more residues of fatty acids selected from the group consisting of myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoleidic acid, alpha-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, and docosahexaenoic acid.

[0097] In some embodiments, the residue is the remainder of a precursor having a linear or branched alkyl chain supporting one or more acidic groups and at least one ethylene group (carbon-carbon double bond). Preferably, at least one such ethylene group is a terminal ethylene group. The terminal ethylene group results in improved reactivity compared to non-terminal ethylene groups during the formation of the elastomer bulk in the manufacturing process. A linear chain having one terminal ethylene group and one acidic group, such as a carboxyl group, is a preferred example. In such a case, there may be 5 to 15 carbon atoms in the chain.

[0098] In some embodiments, the elastomer bulk comprises a silicone or polydiene backbone. Polybutadiene and polyisoprene are examples of polydienes. Polydimethylsiloxane is an example of a polysiloxane (silicone). Because silicones have relatively high water permeability and transparency, optical inspection of cell cultures can be relatively easy.

[0099] In some embodiments, residues are covalently bonded to the elastomer bulk as a result of a reaction between the unsaturated carbon-carbon bond of the residue precursor and the vinyl or hydride functional group of the elastomer bulk. In polydienes, such a reaction is with a vinyl group, while in silicones, such a bond is typically with a silyl hydride functional group.

[0100] More generally, the term “biocompatible” as used herein refers to a material capable of enabling the growth of cells on it.

[0101] The term "more biocompatible material" refers to a material on which the degree of cell proliferation is higher than that of a given material being compared to that "more biocompatible material."

[0102] In some embodiments, the support member 106 is formed from a material that is more biocompatible than the material forming the substrate 102, so that cells and / or tissues preferentially adhere to the support member 106.

[0103] For example, the support member 106 may be formed of silicone, for example, silicone modified by the method described above. On the other hand, at least a portion of the substrate 102 may be formed of a polymer material with lower affinity to biomaterials, for example, a polymer material comprising one or more materials selected from the group consisting of polycarbonate, polyetheretherketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and polyethersulfone (PES). For example, the side wall 108 of the chamber unit 104 may be formed of a polymer material with lower affinity to biomaterials, for example, polycarbonate, polyetheretherketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and / or polyethersulfone (PES).

[0104] In at least some embodiments, the support member 106 includes an elastomer material (which may also be referred to herein as rubber or rubber material), for example, an elastomer material comprising one or more materials selected from the group consisting of silicone and polybutadiene. The elastic properties of such an elastomer material, coupled with the fact that such a material is biocompatible or can be modified to provide at least sufficient biocompatibility, make such an elastomer material particularly suitable for inclusion in the support member 106.

[0105] Alternatively, or in addition, the support member 106 is formed from a material that is more flexible, for example, has a lower Shore A hardness, and / or is more pliable than the material forming the base material 102. Thus, the mechanical properties of the support member 106 are suitable for supporting biomaterials cultured in the chamber unit 104, such as cells or tissues, while the base material 102 contributes more to the structural rigidity of the device 100.

[0106] In embodiments in which the support member 106 includes silicone, for example, is formed from silicone, it should also be noted that the silicone support member 106 is optically transparent and exhibits limited autofluorescence, so that various conventional optical inspection techniques and protocols, with or without staining, can be used in inspections performed using the device 100.

[0107] The selection of materials for the base material 102 and the support member 106 will be discussed in more detail below in relation to the method 10 for manufacturing the device 100.

[0108] The following is a list of example materials for the support member 106.

[0109] In some embodiments, referring again to Figures 1A and 1B, the support member 106 includes at least one structural feature 112 that contacts the biomaterial received within the chamber unit 104. Such a structural feature 112 provides a shape or profile suitable for growing cells or tissues. The shape or profile of the structural feature 112 is selected, for example, depending on the type of cells or tissues to be grown and / or the examination to be performed on the biomaterial.

[0110] In some embodiments, such as those shown in Figures 1A, 1B, 2A, and 2B, at least one structural feature 112 comprises, for each chamber unit 104, a pair of flexible projections, such as flexible posts or pillars, that support tissue on top of and between. In such embodiments, the device 100 is used to examine muscle tissue, such as cardiac tissue, that has grown between and around the pair of projections. As the muscle tissue contracts, the pair of projections bend toward each other, and the resulting displacement of the projections can be measured to determine one or more contractile properties, such as the degree of contraction, the frequency of contraction, and / or the contractile force produced by the tissue contraction.

[0111] Such measurements are described, for example, in International Application No. 2022 / 112291, the entirety of which is incorporated herein by reference.

[0112] The protrusions may have any suitable height as long as they can perform their tissue inspection function. In some embodiments, referring to Figure 2B, each protrusion has a height H or maximum dimension of 500 to 1500 μm, preferably 600 to 1000 μm.

[0113] Such a height H, in conjunction with the fact that the thickness of the support member 106 between its outer surface and the inner surface 116 where the protrusion originates is at most 4 mm, helps in the analysis of the tissue by optical microscope through the support member 106, provided that the support member 106 is optically transparent at least below the protrusion.

[0114] Alternatively, or in addition, each projection has a width W of 100 to 300 μm and / or a length L of 400 to 600 μm.

[0115] The spacing SP between protrusions, for example, between protrusions having the above dimensions of height H, width W, and / or length L, may be 800 to 1200 μm.

[0116] It should be noted that the flexibility of the protruding portion is obtained by the fact that at least the protruding portion of the support member 106 contains an elastomer material, such as one or more materials selected from the group consisting of silicone and polybutadiene.

[0117] In some embodiments, as best shown in Figures 2A and 2B, the support member 106 has a recess 114 defined for each chamber unit 104. A base surface 116 that demarcates part of the recess 114 is joined to the main surface 118 of the support member 106 by a lateral surface 120 that extends between the base surface 116 and the main surface 118.

[0118] Structural features 112, such as flexible projections, are located on and / or within the base surface 116 of the recess 114. Alternatively, or in addition, the recess 114 may be elongated or, for example, elliptical when viewed from above, as shown in the upper region of Figures 1B and 2B.

[0119] In the case of such an elongated recess 114, and when the protrusion is included in the support member 106, the protrusion is positioned along the longitudinal axis 122, which is the direction in which the recess 114 lengthens. Contraction of the muscle tissue between the protrusions occurs along this longitudinal axis 122.

[0120] The recess 114, for example, an elongated recess 114, can have any suitable dimensions. In some embodiments, referring to Figure 2B, the recess 114 has a width W1 of 1500 to 2500 μm, preferably 1800 to 2200 μm, and / or a length L1 of 2500 to 3500 μm, preferably 2700 to 3200 μm. In embodiments where the recess 114 is elongated, the length L1 extending along the longitudinal axis 122 is obviously greater than the width W1.

[0121] The height H1 of the recess 114, for example, the recess 114 having the dimensions of the width W1 and / or length L1 described above, is 500 to 1500 μm, preferably 600 to 1000 μm.

[0122] It should be noted that the height H1 of the recess 114 may be the same as the height H of the protruding part if present, or may be within 10% of that height.

[0123] In some embodiments, it should be noted that structural features 112, such as protrusions, are included in the support member 106, in which case no recesses 114 are defined in the support member 106.

[0124] In this embodiment, the side walls 108 of the chamber unit 104 are used to provide appropriate lateral containment of biomaterials, cell culture media, etc., within each of the chamber units 104.

[0125] In some embodiments, such as those shown in Figure 2A, at least one chamber unit 104, for example, each of the chamber units 104, is provided with a central portion 124 of a support member 106, from which a plurality of rib elements 126 extend, each engaging with the side wall 108 of the corresponding chamber unit 104. Such rib elements 126 contribute to reducing or minimizing deformation of the central portion 124 while maintaining shear stress of a size sufficient to help maintain the support member in place.

[0126] Any suitable number of rib elements 126 can be intended, such as two, three, four, five (as shown in Figure 2A), or more. Preferably, a symmetrical arrangement of rib elements 126 is employed, in which two rib elements 126 are arranged along a first axis 128 and two rib elements 126 are arranged along a second axis 130 perpendicular to the first axis 128.

[0127] It should be noted that the recess 114 is defined in the central portion 124 of the support member 106, with, for example, a structural feature 112, such as a flexible projection, positioned on and / or within the base surface 116 of the recess 114. In the case of an elongated recess 114, the longitudinal axis 122 described above may coincide with the axis 128, which is the direction in which the rib element 126 extends to reach the side wall 108 of the chamber unit 104.

[0128] In some embodiments, the support member 106 is porous so that it is permeable to fluids, such as gases. Such a porous support member 106 can be obtained by any suitable method, for example, by forming openings in the material constituting the support member 106, such as an elastomer material. Such openings can be formed, for example, by laser ablation or casting imprint drilling.

[0129] Laser drilling / ablation is described, for example, in U.S. Patent Publication No. 2014 / 127744 and U.S. Patent Publication No. 2018 / 0315409. Cast imprint drilling is described, for example, in U.S. Patent Publication No. 2020 / 360923, U.S. Patent Publication No. 2015 / 010919 and U.S. Patent Publication No. 2022 / 228108, all of which are incorporated herein by reference in their entirety.

[0130] Each opening is preferably sized to restrict or prevent cells supported on the support member 106 from passing through it. For this purpose, each opening has a diameter of less than 10 μm.

[0131] In general, bonding substrates 102 and support members 106 made of different materials, as disclosed herein, to each other in devices such as those described with reference to Figures 1A-1D and Figures 2A and 2B presents various technical challenges. It is difficult to perform such bonding in a sterile manner and / or without unintentionally creating areas between the substrate 102 and support members 106 where fluid leakage that may interfere with use may occur. Furthermore, additional manufacturing steps are required to bond the substrates 102 and support members 106 to each other, increasing manufacturing complexity and cost. Moreover, in embodiments in which the support member 106 includes a flexible projection, the substrate 102 and support member 106 need to be aligned so that the projection is properly oriented, for example, within each chamber unit 108 of a plurality of chamber units 108.

[0132] For these reasons, at least again referring to Figures 1A and 1B, the method 10 according to the present disclosure has a step 12 of using a mold to mold one of the base material 102 and the support member 106 onto at least a portion of the other of the base material 106 and the support member 102, wherein at least a portion of the other of the base material 106 and the support member 102 is positioned in or adjacent to the mold during the molding step 12. For example, in the embodiment schematically depicted in Figures 1A and 1B, the support member 106 is molded onto the base material 102.

[0133] Step 12 involves molding one of the base material 102 and the support member 106 onto at least a portion of the other of the base material 106 and the support member 102, thereby shaping the former and simultaneously bonding it to the latter. In this way, a separate assembly step of bonding the base material 102 and the support member 106 to each other can be omitted. In particular, no adhesive or glue is required to keep the base material 102 and the support member 106 bonded to each other.

[0134] Method 10 provides a relatively rapid and reliable process for manufacturing the device 100, in which the substrate 102 and the support member 106 can be considered as a single integrated piece. Furthermore, the support member 106, for example, a silicone support member 106, is fixed by its shape within a frame defined by the substrate 102, for example, a polycarbonate frame. In this way, a portion of the substrate is embedded in a portion of the support member such that one cannot be easily separated from the other.

[0135] Thus, the adhesion, such as leak-free adhesion, achieved by step 12 in which one of the base material 102 and the support member 106 is molded onto the other of the base material 106 and the support member 102 is supplemented by morphological fixation, particularly in embodiments in which the support member 106 comprises a central portion 124 and rib elements 126 that engage with the side wall 108 of the chamber unit 104. In other embodiments, the central portion 124-rib element 126 configuration can be omitted, for example, to simplify the mold design.

[0136] It should also be noted that, at a microscopic scale, surface irregularities of the first molded part (e.g., substrate 102) (e.g., irregularities at the molecular scale) become at least partially conformal to the second molded part (e.g., support member 106) due to the second molded part being molded on top of the first molded part. This contributes to improved adhesion between the two parts of a device fabricated using the method disclosed herein, benefiting its leak characteristics. The method of molding one part on top of the other thus allows for the use of irregularly shaped contact surfaces between the substrate and the support member, which would be more difficult to achieve good contact by other methods. For example, separately fabricated components having contact surfaces with complex irregular shapes that are complementary and conform to each other to make close contact are difficult or more expensive to fabricate. For this reason, inspection of the contact surfaces between the substrate and the support member of the device disclosed herein is used to distinguish the device disclosed herein from other multi-component devices not fabricated using the method disclosed herein, such as methods utilizing clamping, bonding, or gluing of separately fabricated parts. Co-form adhesion may also be observed in components that are glued together, in which case the additional layer of adhesive helps to distinguish such glued devices from devices fabricated using the methods disclosed herein.

[0137] The molding step 12 implies that one of the substrate 102 and the support member 106 is able to withstand the molding conditions, with the other being molded on top of the other. This affects one or more of the following: which of the substrate 102 and the support member 106 is molded on top of the other (or conversely, which component is prefabricated before the molding step 12), the material and / or precursor selected to form the substrate 102 and the support member 106, and the conditions in the molding step 12.

[0138] In at least some embodiments, the molding step 12 is defined, for example, by injection molding, which includes injection molding one of the base material 102 and the support member 106 onto at least a portion of the other of the base material 106 and the support member 102. Injection molding provides a cost-effective and scalable method for manufacturing the device 100.

[0139] In some embodiments, such as those shown in Figure 3, method 10 includes step 14 of releasing the substrate 102 together with the support member 106 from the mold while the temperature of the substrate 102 and the support member 106 is higher than room temperature, preferably at least 40°C, for example, 40 to 80°C. This minimizes the risk of one or both of the substrate 102 and the support member 106 becoming stuck to the mold.

[0140] In some embodiments, continuing with reference to Figure 3, the step 12 of molding one of the substrate 102 and the support member 106 onto at least a portion of the other of the substrate 106 and the support member 102, for example, the step of injection molding, includes curing the precursor material 12B. In the case of injection molding, the molding step 12 includes injecting the precursor material into a mold 12A, and then curing the precursor material in the mold and on the other of the substrate 106 and the support member 102 12B.

[0141] In this embodiment, curing 12B includes heating the precursor material to a temperature higher than, for example, 160°C, for example, 160-200°C.

[0142] Heating at such temperatures helps ensure the curing of the precursor material and its adhesion to the initial molded components.

[0143] After heating, the base material 102 and the support member 106 are cooled to room temperature, but as already described, it is preferable that the base material 102 is released from the mold together with the support member 106 before the base material 102 and the support member 106 have cooled to room temperature (14).

[0144] It should be noted that the process of thermally curing the precursor material 12B requires the selection of a thermally robust material suitable for one of the substrate 102 and the support member 106, on which the other of the substrate 106 and the support member 102 will be molded via the curing process 12B.

[0145] For this purpose, the glass transition temperature of the material forming one of the base material 102 and the support member 106, on which the other of the base material 106 and the support member 102 is molded, is at least 140°C.

[0146] This minimum glass transition temperature allows for the curing 12B of the precursor material at a relatively high temperature during the molding step 12.

[0147] Examples of materials having a glass transition temperature of at least 140°C include polycarbonate, polyether ether ketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), polyethersulfone (PES), and combinations of the above polymers, such as a mixture or alloy of at least two of polyether ether ketone (PEEK), polyetherimide (PEI), and polyethersulfone (PES).

[0148] In embodiments in which the support member 106 contains silicone, for example, is formed from silicone, curing the precursor material 12B includes curing the siloxane precursor material.

[0149] Such curing 12B is carried out, for example, at 140°C to 180°C. Alternatively, or in addition, a precursor material, such as a siloxane precursor material, is injected into the mold at a pressure of 100 to 600 bar, preferably when the mold temperature is above 110°C.

[0150] In some embodiments, such as those shown in Figures 1A, 1B, and 3, Method 10 comprises a step 16 for initial molding of the substrate 102 or the support member 106, for example, an initial injection molding step 16, and a step 12, 12A, 12B for molding the unmolded substrate 102 or support member 106 onto the initial molding component, for example, a subsequent injection molding step 12, 12A, 12B. Thus, Method 10 has a two-stage molding process, for example, a two-stage injection molding process.

[0151] The initial molding step 16 includes injection molding 16 of a thermoplastic, for example, one or more selected from polycarbonate, polyetheretherketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and polyethersulfone (PES). Such a thermoplastic is melted at a temperature of, for example, 200-300°C and injected into a mold at a pressure of 600-2000 bar, preferably at a temperature below the glass transition temperature of the thermoplastic. For example, the mold temperature is 80-120°C.

[0152] In some embodiments, referring to Figure 3, Method 10 includes steps 18A, 18B to adjust the mold, from a first configuration used in a step 16 for initial molding of the substrate 102 or support member 106, for example in a step 16 for initial injection molding, to a second configuration used in steps 12, 12A, 12B for molding the unmolded of the substrate 102 and support member 106 onto the initial molding configuration, for example in subsequent injection molding steps 12, 12A, 12B.

[0153] The mold includes a first molded component and a second molded component, and the base material 102 or the support member 106 is moldable between the first molded component and the second molded component. The mold further includes a third molded component, and the component of the base material 102 and the support member 106 that is not moldable between the first molded component and the second molded component is moldable between the third component and the component molded between the first molded component and the second molded component.

[0154] In such embodiments, steps 18A, 18B for adjusting the mold from a first configuration to a second configuration include switching the second mold part to a third mold part while the components molded between the first and second mold parts remain received within the first mold part. Such switching includes, for example, displacing the second mold part to open the mold 18A, and then closing the mold again using the third mold part 18B.

[0155] Step 16 for initial molding of the base material 102 or support member 106, for example, step 16 for injection molding, includes heating the material to a temperature higher than room temperature to make the material moldable, for example, injectable into the space between the first molded part and the second molded part described above.

[0156] In such embodiments, step 12, which subsequently involves molding the unmolded substrate 102 and support member 106 onto the initial molded component, is performed before the initial molded component returns to room temperature, preferably while the initial molded component is at least 80°C. This contributes to bonding the substrate 102 and support member 106 to each other. In embodiments where the subsequent molding step 12 includes curing the precursor material, maintaining such a high temperature may also contribute to such curing.

[0157] In some preferred embodiments, the substrate 102 is initially molded from a thermoplastic material having a glass transition temperature of at least 140°C, such as polycarbonate, polyetheretherketone (PEEK), acrylonitrile butadiene styrene (ABS), polyetherimide (PEI), and / or polyethersulfone (PES) 16, and the support member 106 is subsequently molded (12, 12A, 12B) onto the substrate 102 via curing a precursor material, such as a siloxane precursor material 12B.

[0158] In this embodiment, one side of the mold is opened (18A) while the initially molded substrate 102 remains inside the mold. Subsequently, a third mold component closes the mold to form a second mold configuration, into which a precursor material, such as a siloxane precursor material, is injected.

[0159] In the second configuration, the mold is maintained at a temperature of 80-120°C, and it should be noted that while the mold is at this temperature, the precursor material, such as a siloxane precursor material, is injected.

[0160] Once filled with a precursor material, such as a siloxane precursor material, the mold is heated to 160-200°C until the precursor material hardens, a support member 106 is formed, and the support member 106 adheres to the substrate 102, for example, to the thermoplastic of the substrate 102.

[0161] From a more general perspective, the elastomer material is molded while in the mold, in at least partial contact with the surface of a substrate 102, such as a thermoplastic substrate 102.

[0162] The mold is then cooled to, for example, 40-80°C, at which point the two-component product is released from the mold. This reduces the risk of the two-component product adhering to the mold, as previously explained.

[0163] The molding method disclosed herein has been described with reference to the devices shown in Figures 1A-D and Figures 2A and 2B. However, the method is not limited to the manufacture of such devices and can be used for many other devices for various biological testing purposes. Some examples are given below.

[0164] In embodiments where several chamber units 104 are defined on the substrate 102, the chamber units 104 may be arranged, again, to form multiple rows 105. In some embodiments, such as those shown in Figure 4, twelve rows 105 of chamber units 104 are defined on the substrate 102, each row comprising eight chamber units 104, similar to the rows of chamber units 104 shown in Figures 1A and 1B.

[0165] Therefore, the device 100 has 96 chamber units 104, for example, a well configuration. In other embodiments, the device 100 has 24 chamber units 104, for example, a well configuration. The number of chamber units 104 defined in the substrate 102 corresponds to the number of wells, size, and shape of well plates that are well known and commonly used in the art, along with the shape and size of the substrate 102 and, for example, the device 100 as a whole.

[0166] Furthermore, in some embodiments such as the embodiment shown in Figure 4, the support member 106 has a recess 114 defined for at least some of the chamber units 104, for example, each of them, and / or the support member 106 has at least one structural feature 112, for example, a flexible projection, that contacts the biomaterial received in at least some of the chamber units 104, for example, each of them.

[0167] Figures 5A to 5C provide various diagrams of the biomaterial testing device 100, which has a substrate design similar to that shown in Figure 4, but with a support member 106 having the end portion 110 described above in relation to Figure 2A for each chamber unit 104. The molding step 12 allows the substrate 102 and the support member 106 to adhere to each other, as already described. Nevertheless, in the exploded perspective view provided in Figure 5A, the substrate 102 and the support member 106 are shown separated from each other for clarity, whereas in the perspective view of the device 100 provided in Figure 5B and the plan view provided in Figure 5C, the substrate 102 and the support member 106 are shown adhered to each other.

[0168] In some embodiments, such as those shown in Figure 6, the biomaterial testing device 100 includes a base member 132, such as a glass base member 132, on which a support member 106 and a substrate 102 are placed.

[0169] The base member 132 contributes to the reinforcement of the device 100. Alternatively, or in addition, the base member 132, for example, a glass base member 132, is optically transparent to allow for the analysis of biomaterials through the glass base member 132 using an optical microscope.

[0170] The base member 132, for example, the glass base member 132, also provides a smooth surface on which, in some embodiments, the support member will be molded. Such a smooth surface consequently contributes to the outer surface of the support member 106 having a smooth surface. This facilitates the analysis of biomaterials through the support member 106 using an optical microscope.

[0171] It should be noted that in other embodiments, where the base member 132 is not included in the device 100, or more generally, where the outer surface of the support member 106 is in contact with the surface of the mold during manufacturing, the surface of the mold is preferably a polished surface.

[0172] Such polished surfaces contribute to providing a relatively smooth outer surface of the support member 106, which ultimately facilitates the analysis of biomaterials through the support member using an optical microscope.

[0173] It should be noted here that the focal length of many optical microscopy setups, for example, those used to image stains, can be as short as approximately 0.6 mm. The distance from the bottom of the base member 132, for example, the glass base member 132, to the support member 106 may be 0.6 mm or less accordingly. However, other thicknesses may be used for different optical setups. More details on such optical setups and options for the material and design of the base plate are disclosed in currently unpublished pending patent applications, European Patent Application No. 23181319.7 and PCT / EP2024 / 066936, and European Patent Application No. 23182831.0 and PCT / EP2024 / 067028, all of which are incorporated herein by reference in their entirety, although other devices are also possible.

[0174] The thickness of the optically transparent base member 132, for example, the glass base member 132, is 0.10 to 0.30 mm, for example, about 0.20 mm.

[0175] Such a thickness takes into account the thickness of the support member 106, including, for example, the height H of the flexible protrusion included in the support member 106.

[0176] More generally, method 10 includes the steps of placing at least a portion of the base member 132 inside or adjacent to the mold, and the step 16 of initial molding the substrate 102 or the support member 106 on the base member 132. Following the step 12 of molding whichever of the substrate 102 and the support member 106 has not yet been molded onto the initial molding component, the base member 132 on which the substrate 102 and the support member 106 have been molded is removed from the mold, for example, demolded.

[0177] In some embodiments, such as those shown in Figures 7 and 8, at least one double chamber unit 104 is defined on the substrate 102, each of which comprises a first chamber 134 and a second chamber 136, fluidly connected to each other via a bridge 138 (sometimes called a top fluid channel) defined between the first chamber 134 and the second chamber 136. Furthermore, the double chamber units 104 can be interconnected in series, parallel, or mixed configurations within a single device by fluid channels. For example, in Figure 8, multiple double chamber units in each row 105 are interconnected in parallel between a first channel 137 and a second channel 139. The first channel may be called a fluid supply channel, and the second channel may be called a fluid discharge channel. However, as mentioned above, other configurations can also be specified. In the devices of Figures 7 and 8, the fluid channels 137, 138, and 139 are part of the substrate 102. These may also be called top channels of the device.

[0178] Such a dual-chamber unit 104 is useful for studying cancer cell metastasis. It is possible to evaluate cells released or expelled by any cell culture provided in the first chamber 134. The first chamber 134 can therefore be used to treat the cell culture, for example, by administering a specific therapeutic solution or drug, such as a chemotherapy solution, and the second chamber 136 captures any released cells to analyze the effect of the treatment on metastasis performed in the first chamber 134.

[0179] In such embodiments, the support member 106 comprises, for example, the porous support member 106 described above for supporting a biomaterial, such as cells, received within the double chamber unit 104. The support member 106 includes, for example, a first porous portion for supporting a biomaterial, such as cells, within a first chamber 134, and a second porous portion for supporting a biomaterial, such as cells, within a second chamber 136.

[0180] Any suitable number of double chamber units 104 can be defined on the base material 102; for example, two double chamber units 104 in the embodiment shown in Figure 7, and 24 double chamber units 104 in the embodiment shown in Figure 8.

[0181] Further examples and more detailed designs of such double-chamber unit devices are described in unpublished patent applications, European Patent Application No. 23181319.7 and PCT / EP2024 / 066936, which are incorporated herein by reference in their entirety.

[0182] The base material 102 and support member 106 of the device 100 having such a double chamber unit can be formed using method 10 according to any embodiment described herein.

[0183] More generally, method 10 can be used to form any device for examining a biomaterial having at least a substrate and a support member, as defined herein. Several examples of such devices are disclosed in European Patent Application No. 23181319.7 and PCT / EP2024 / 066936, and European Patent Application No. 23182831.0 and PCT / EP2024 / 067028, all of which are incorporated herein by reference in their entirety, but other devices are also possible.

[0184] Figures 9A–9F are used to provide examples of how to apply Method 10 to fabricate devices having one or more top channels and / or one or more bottom channels, such as the device in Figure 8 which has top channels 137, 138, and 139, as well as other features within the device. Figures 9A–9F also serve to illustrate mold systems having various parts that will be used in the Method. The example drawings in Figures 9A–9G show a substrate and support member having two chamber units, but these drawings are also thought to represent cases where there are more such chamber units, such as in the devices in Figures 5A–5C, 6, and 8.

[0185] Figures 9A to 9F illustrate a method for manufacturing a three-component multi-chamber device 900 having parallel-connected chamber units very similar to the device in Figure 8, the difference being that, compared to the chamber units in Figures 7 and 8, device 900 has only one chamber (or container) per chamber unit.

[0186] Figure 9E shows a cross-sectional view of the device 900 to be fabricated, and Figures 9F and 9G show the bottom and top views of the device 900, respectively. Figure 9E shows three cross-sectional views of the device 900. The two lower figures, IX and X, show cross-sections perpendicular to the cross-section in the upper figure, along directions IX and X shown in the upper figure.

[0187] The device 900 comprises two chamber units 934 within a base material 902, each having a cylindrical shape with a diameter of 170. Other shapes can be used. In Figure 9F, the perimeter of the chamber sidewall is shown with a dashed line because, in this figure, the chamber cannot be seen from the bottom side of the device 900 through the support member 906.

[0188] The device further comprises two top channels 937 and 939 extending horizontally from the substrate 902 in the top view of Figure 9G. The chamber unit 934 is connected to channels 937 and 939 via channel 938 so that the chambers are in a parallel configuration.

[0189] A support member 902 is present on the bottom side of the device 900, which is bonded to the substrate 902 according to the principles disclosed herein.

[0190] The support member 906 includes a bottom channel 935 that connects the sub-chambers 975 located below the membrane 942. In the bottom view of Figure 9F, the channel 935 can be seen extending horizontally.

[0191] The support member 902 has ends 910, each of which is located within one of the chamber units 934. Each end includes a film 942 having a circular shape with a diameter of 972 and a thickness measured perpendicular to the extending surface. In Figures 9F and 9G, the circular shape can be viewed from above or below. Each end also has a flange 966, each of which extends upward from the bottom side of the base material 902 into the chamber unit 934 to a height less than the height of the chamber unit 934. One of each film 942 extends from the flange 966, or in other words, is supported on or integrated with the flange.

[0192] The preferred film thickness is in the range of 5 to 100 μm, but other thicknesses can be used. This can be selected according to the desired purpose of the device.

[0193] In some embodiments, the membrane comprises or consists of a perforated membrane having multiple pores. Preferably, each pore is wide enough to allow a fluid that may contain nutrients, drugs, or other (bio)molecules such as proteins to pass through.

[0194] Depending on the application, the pores may be large enough to allow, at least partially, the passage of cells or a specific type of cell.

[0195] In some embodiments, the pores are narrow enough to prevent cells from passing through. For example, the pore diameter is in the range of 10 μm or less, for example, 5 μm. However, other values, as described below herein, may be used. These allow a porous membrane to support tissue samples such as cells, while the entire contents of the membrane can be supplied to the cells by passing the fluid described above through one or more channels in the device. Exemplary membranes and channels are seen in the devices of Figures 7 and 8, and in the devices described, for example, in European Patent Application No. 23181319.7 and PCT / EP2024 / 066936.

[0196] The support member 902 may have other structural features 912 at its end 934. In this case, for example, the membrane may have multiple wells defined. The bottoms of such wells also form a membrane or perforated membrane as described herein. Detailed examples of membranes including such wells are described in European Patent Application No. 23181319.7 and PCT / EP2024 / 066936.

[0197] The pores are fabricated, for example, using laser drilling as disclosed herein.

[0198] Figures 9A to 9D show three cross-sectional views of a mold system as defined herein, used in the manner defined herein for manufacturing device 900. It will be understood that the mold system can be used for manufacturing other devices by adjusting its structural design. The mold system comprises a first mold part 950 and a second mold part 952, which can be combined to form a first configuration for the initial molding step 16 as defined herein. The first and second mold parts have shapes and sizes designed to mold the substrate 902 of device 900. The two lower figures I and II show cross-sections perpendicular to the cross-section in the upper figure, along the directions indicated in I and II in the upper figure.

[0199] The first and second molded parts are geometrically designed and shaped together to obtain protrusions and open spaces (sometimes referred to as one or more first open spaces) that define the shape and design of the substrate 902 during and after molding. In this case, the first and second molded parts have protrusions 953 that define at least partially the chamber unit 934 and protrusions 955 that ultimately define the top channel of the substrate 902 (very similar to channels 137 and 139 of the device in Figure 8), which will be defined by filling the open space 954. More specifically, the protrusions 953 are cylindrical in this case with a diameter of 170, their cylindrical axis is vertical and in the plane of the cross-sectional view. After molding, these will ultimately define a cylindrical chamber unit 934 with a diameter of 170 on the substrate 902. The protrusions 955 are rectangular and extend along the first mold perpendicular to the drawing plane of the cross-sectional view I. The first molded part 950 shows that a projection 957 is in contact with a projection 953. The projection 957 is rectangular and extends along only a portion of the cylindrical projection 953, which will eventually form a top channel connecting the chamber 934 to the top channel. The second molded part 952 has a surface molded surface portion 980 (sometimes called the second molded surface) that defines a substrate surface 982 which will later define a portion of the substrate on which the support member 906 will be molded. These surfaces 982 will consequently become contact or bonding surfaces as already discussed herein. The molded surface portion 980 is geometrically designed to improve or enhance the adhesion of the support member to the substrate. For clarity, specific geometric designs are not shown in Figures 9A to 9G, but the surface 980 of the first molded part may have, for example, a specific surface roughness and / or a certain type of corrugation.

[0200] Although the first and second mold components are shown separated from each other in an open configuration in Figure 9A, they can be releasably attached to each other (separable) in a leak-free manner, and in the first mold configuration (where they are combined and held together) as shown in Figures 9B and 9D, they can be used to perform molding by injecting liquid material into them. For example, they can be clamped together to form a closed first mold configuration. The first mold component has an injection opening 156 for introducing liquid substrate material into the mold when it is in a closed molding configuration.

[0201] In the initial molding step 16, described with reference to Method 10 and Figure 3, the first molded part 950 and the second molded part 952 are put into a first configuration, and the base material is molded into the base material 902 by injection molding the material into the mold opening 954, for example, through the injection opening 156, under the conditions described. The result is shown in the cross-sectional view of Figure 9B. As an example, the material to be injected is selected from the group of thermoplastic materials such as polycarbonate, as described above in this specification.

[0202] In a further step 18A, the second molded part 952 is released from the first molded part 950 and the molded base material 902, while the base material 902 remains inside the first molded part 950. The result is shown in Figure 9C. The design of the protrusions 953 of the first molded part 952 and the second molded part 953 results in an assembly of the first molded part and the base material 902 having open spaces 964 with a diameter of 170, each forming an exposed portion 964 of the base material chamber 134. These open spaces will ultimately be occupied by the end 910 of the support material once molded.

[0203] In step 18B, the second mold part 952 is replaced with a third mold part 962 designed to fit the first mold part 950, forming a second mold configuration in which a further open space (sometimes referred to as one or more second open spaces) remains, into which the support member 906 will be molded. The design of the first mold part 950, the second mold part 960, and the molded base material 902 now defines the further open space. This second mold configuration is shown in Figure 9D.

[0204] The third molded part 962 includes a projection 961 having a cylindrical shape with a diameter of 172 and a cylindrical axis oriented vertically and in the drawing plane. The diameter 972 is smaller than the diameter 970 of the projection 953 of the first molded part 950, and the height of the projections is such that they do not come into contact with the projection 953 of the molded part 950 in the second closed configuration. This leaves each of the chamber units 934 with an open space in the form of an exposed portion 964, which will be occupied by the material that will form the end portion of the base material member 906. In this case, the further open space is therefore designed to result in a support member 906 having a thin film 942 attached to the flange 966, the thin film being between the projections 953 and 961. However, other designs may be used, and the designs of the projections 953 and 961 may be used to define a desired end portion, such as those defined herein. In this context, surface 964 may be referred to as the first mold surface, and surface 986 of the protrusion 961 may be referred to as the third mold surface.

[0205] The third molded part also has one or more protrusions (only one is shown for clarity) 963, which have a rectangular shape and extend perpendicular to the drawing of cross-sectional view VII. These protrusions will ultimately form channels 935 on the support member 906 that interconnect the sub-chambers 975 located below the membrane 942.

[0206] In step 12, when the second mold configuration is taken, the support member 906 is molded as described herein with reference to method 10 and Figure 3. The molding material is injected through the opening 960 to form the result shown in Figure 9D. Since the material of the support member is pressed against a portion of the surface of the substrate and molded, the contact between the contact surfaces is close, and the support member is molded to be substantially conformal to the shape and irregularities of the surface of the substrate 902. As a result, a close contact with improved adhesion and leakage characteristics is obtained, as described herein.

[0207] After cooling to a suitable temperature as already described in relation to method 10 herein, the device in the form of the support member 906 molded on the substrate 902 is demolded from the first and second molded parts. The result is the device 900 shown in Figures 9E to 9F as described above herein. If no further parts are needed, this device can be used as is. This is the case, for example, when the support member has a design that does not have a bottom channel or other opening such as channel 935 that needs to contain fluid. A device having a pillar as described herein is such an example. However, the device in Figure 9E has a bottom channel, which may need to be closed from the bottom side.

[0208] The molding system used in the method described with reference to Figures 9A to 9D comprises a first molding component, a second molding component, and a third molding component as defined herein.

[0209] In the alternative method according to the present invention, the molding of the substrate and the molding of the support member are not combined. In this case, the substrate is an existing substrate, which may be a molded product, but is supplied from stock. The first molded part is designed to correspond to the substrate as defined herein and as supplied from stock. The first and second molded parts are designed and used as described with reference to Figures 9A to 9D. In such an alternative method, the mold system according to the present disclosure then consists only of the first molded part and the third molded part as defined herein.

[0210] To close the bottom channel, the device can be bonded to a base member 932 made of, for example, glass or other suitable material such as, for example, the material described herein. Figure 9H shows the device of Figures 9E to 9G having a base member 932, which closes the channel 935 and the opening 975 at the bottom of the device so that they are suitable for containing fluid. The channel is now suitable for supplying fluid to the membrane 942 from the bottom or for draining fluid that has passed through such membrane from the top to the bottom.

[0211] In the alternative method, before the molding material for the base material member 906 is injected, the base member is already present in the third mold part 962, for example as a fitting material, or in a pre-molded state within the third mold part 962. In this case, the support member is molded to be conformable not only to the base material 902 but also to the base member 932.

[0212] A top cover plate can be added as needed. This is releasably attached to the substrate 902. The top cover plate is made of glass or polymer material. It may be made of the same material as the substrate material. The top cover is not shown in Figure 9H for clarity.

[0213] It will be apparent that two-component or multi-component devices described herein may have desired designs different from the examples provided herein. In this case, the size and shape of the chamber unit 934 can be determined by adjusting the design and dimensions of the protrusions 935 and 961. Furthermore, the design and dimensions can also be used to define an open space between the protrusions 935 and 961, thereby determining the shape and dimensions of the end 910 and the membrane 942. For example, the thickness of the membrane is set using these design parameters. Wells are also defined in the membrane. More detailed designs of membranes with perforations and / or wells, including desired dimensions of the perforations and / or wells, are described in other literature provided herein. Also, device 900 has only one chamber per chamber unit. There may be two or more chambers per chamber unit, as in device 800 in Figure 8. Figure 9 shows device 900 having only two chambers and corresponding channels. However, this is merely illustrative, and the principles shown can be extended to the manufacture of devices having different numbers of chambers and different numbers of channels, etc. Therefore, for example, 96 chamber plates can be manufactured using this method.

[0214] More generally, this disclosure proposes the use of device 100 according to any of the embodiments described herein for examining biomaterials, such as cell populations, double cell layers, spheroids, organoids, or biopsy specimens.

[0215] This disclosure further envisions a drug testing method comprising the steps of providing a biomaterial into at least one chamber unit 104 of a device 100 according to any embodiment described herein, and exposing the biomaterial to a drug to be tested.

[0216] In some embodiments, the step of providing biomaterial into at least one chamber unit 104 includes culturing cells in at least one chamber unit 104.

[0217] Alternatively, or in addition, a drug testing method may include a step of monitoring the reaction of a biological material, such as cultured cells, to the drug to be tested. This monitoring step may be carried out in any suitable manner, for example, by an optical microscope, or through the support member 106 if the support member 106 is optically transparent.

[0218] Preferred combinations of silicone and (rigid) thermoplastics are silicone / polycarbonate, or silicone / PEEK, and the following mixtures of silicone / PEEK, when the glass transition temperature (Tg) of polycarbonate is greater than 140°C:

[0219] [Table 1]

[0220] The following exemplary commercially available elastomers / flexible materials (which are transparent or translucent and approved for food contact) may be used as the material for the support member 106 in certain embodiments: 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, CELLENEMC3261|TPE, Dynaflex (trademark) G2706-1000-00|TPE, Dynaflex (trademark) G2711-1000-00|TPE, Elastocon (registered trademark) 2860L|TPE, Filter-bond (trademark) E-3264|TS, FLEXCHEM (trademark) 3551-02|PVC, flexible, FLEXCHEM (trademark) 4051-02|PVC, flexible Flexible, FLEXCHEM (trademark) 4551-02 | PVC, Flexible, FLEXCHEM (trademark) 5051-02 | PVC, Flexible, FLEXCHEM (trademark) 5551-02 | PVC, Flexible, FLEXCHEM (trademark) 6051-02 | PVC, Flexible, FLEXCHEM (trademark) 6551-02 | PVC, Flexible, Medalist (registered trademark) MD-12130 | TPE, Medalist Medallist (registered trademark) MD-12130H|TPE, Medallist (registered trademark) MD-12140|TPE, Medallist (registered trademark) MD-12140H|TPE, Medallist (registered trademark) MD-12150|TPE, Medallist (registered trademark) MD-12150H|TPE, Medallist (registered trademark) MD-12f150S|TPE, Medallist (registered trademark) MD-12160|TPE, Medallist (registered trademark) MD-12160H|TPE, Medallist (registered trademark) MD-12170|TPE, Medallist (registered trademark) MD-12170H|TPE, Medallist (registered trademark) MD-12243|TPE, Medallist (registered trademark) MD-12337|TPE, Medallist (registered trademark) MD-12340NAT|TPE, Medalist(registered trademark) MD-12342|TPE, Medalist(registered trademark) MD-12344|TPE, Medalist(registered trademark) MD-12350|TPE, Medalist(registered trademark) MD-12352|TPE, Medalist(registered trademark) MD-12362|TPE, Medalist(registered trademark) MD-125|TPE, Medalist(registered trademark) MD-130|TPE, Medalist(registered trademark) MD-13240|TPE, Medal ist(registered trademark)MD-135|TPE, Medalist(registered trademark)MD-145|TPE, Medalist(registered trademark)MD-155|TPE, Medalist(registered trademark)MD-17365|TPE, Medalist(registered trademark)MD-225|TPV, Medalist(registered trademark)MD-32045|TPE, Medalist(registered trademark)MD-32245|TPE, Medalist(registered trademark)MD-36048|TPE, Medalist(registered trademark)MD-37063 NAT|TPE, Medalist(registered trademark)MD-42245 XRD1|TPE, Medalist(registered trademark)MD-42245 XRD3|TPE, Medalist(registered trademark)MD-74357 XRD1|TPE, Medalist(registered trademark)MD-74357XRD2|TPE, Mediprene(registered trademark) 500120M|TPE, Mediprene(registered trademark) 500200M|TPE, Mediprene(registered trademark) 500250M|TPE, Mediprene(registered trademark) 500300M|TPE, Mediprene(registered trademark) 500350M|TPE, Mediprene(registered trademark) 500400M|TPE, Mediprene(registered trademark) 500434M|TPE, Mediprene(registered trademark) 500450M|TPE, Mediprene(registered trademark) 500484M|TPE, Medi prene(registered trademark) 500520M|TPE, Mediprene(registered trademark) 500534M|TPE, Mediprene(registered trademark) 500584M|TPE, Mediprene(registered trademark) 500600M|TPE, Mediprene(registered trademark) 500634M|TPE, Mediprene(registered trademark) 500650M|TPE, Mediprene(registered trademark) 500684M|TPE, Mediprene(registered trademark) 500700M|TPE, Monprene(registered trademark) RG-10160H|TPE, ProvaMed(registered trademark) TPE 1120|TPE, ProvaMed(registered trademark) TPE 1160|TPE, RABALON(registered trademark) PJ4300C|TPE, RABALON(registered trademark) PJ5300C|TPE, RABALON(registered trademark) PJ6300C|TPE, RABALON(registered trademark) PJ7300C|TPE, SkinFlex 15 F-115 A / B|TSU, SkinFlex BR-60, BRUSHABLE A / B|TSU, T-Blend(registered trademark) TPE-F22|SEBS, THERMOLAST(registered trademark) M TM3LFT(Series: MC / LF)|TPE, THERMOLAST(registered trademark) M TM3MED(Series: MC / tl)|TPE, THERMOLAST(registered trademark) M TM3RST(Series: MC / RS)|TPE, THERMOLAST(registered trademark) M TM4LFT (Series: MC / LF) | TPE, THERMOLAST® M TM4MED (Series: MC / tl) | TPE, THERMOLAST® M TM4RST (Series: MC / RS) | TPE, THERMOLAST® MTM5LFT (Series: MC / LF) | TPE, THERMOLAST® M TM5MED (Series: MC / tl) | TPE, THERMOLAST® M TM6LFT (Series: MC / LF) | TPE, THERMOLAST® M TM6MED (Series: MC / tl) | TPE, THERMOLAST® M TM7LFT (Series: MC / LF) | TPE THERMOLAST® M TM7MED (Series: MC / tl) | TPE, UNISOFT SPECIAL® DS-35A-CL-M-01 | SEBS UNISOFT SPECIAL® DS-55A-CL-M-01 | SEBS Versaflex® G2705 N | TPE Versaflex® HC 1100-40 Translucent EU | TPE Versaflex® HC 1348 Natural|TPE, Versaflex(TM) HC MT317|TPE, Versaflex(TM) HC MT555|TPE, Versaflex(TM) OM 1040X-1|TPE, CELLENE MC2248|TPE, CELLENE MC2265|TPE, CELLENE MC3038|TPE, CELLENE MC3050|TPE, CELLENE MC3061|TPE, CELLENE MC3226|TPE, CELLENE MC3239|TPE, CELLENE MC3261|TPE, ChronoPrene(TM) 25A|TPE, ChronoPrene(TM) 40A|TPE (CardioTechInternational, Inc.), Dryflex (registered trademark) 500300S|TPE, Dryflex (registered trademark) 500350S|TPE, Dryflex (registered trademark) 500400S|TPE, Dryflex (registered trademark) 500450S|TPE, Dryflex (registered trademark) 500500S|TPE, Dryflex (registered trademark) 500550S|TPE, Dryflex (registered trademark) 500600S|TPE, Dryflex (registered trademark) 500650S|TPE, Dryflex (registered trademark) 500700S|TPE, Dynaflex (trademark) G2701-1000 -02|TPE, Dynaflex (trademark) G2706-1000-00|TPE, Dynaflex (trademark) G2709-1000-00|TPE, Dynaflex (trademark) G2711-1000-00|TPE, Dynaflex (trademark) G2712-1000-02|TPE, Dynaflex (trademark) G2730|TPE, Dynaflex (trademark) G2755-1000-00|TPE, Dynaflex (trademark) G2755C|TPE, Dynaflex (trademark) G6713-0001|TPE, Dynaflex (trademark) G6713C|TPE, Dynalloy (trademark) GP 7810-60T|TPE, Dynalloy (trademark) GP 7810-70T|TPE, Dynalloy (trademark) OBC8200-BT50|TPE, Estane (registered trademark) 58123 TPU|TPU-Polyester, Evoprene (trademark) 019|SBS, Evoprene (trademark) G 925|SEBS, Evoprene (trademark) G 936|SEBS, Evoprene (trademark) G 942|SEBS, Evoprene (trademark) G 958|SEBS, Evoprene (trademark) G 966|SEBS, Evoprene (trademark) G 967|SEBS, Evoprene (trademark) G 968|SEBS, Evoprene (trademark) G 969|SEBS, Evoprene (trademark) G 970|SEBS, Evoprene (trademark) GC 5685|SEBS, Evoprene (trademark) GC 5686|SEBS, Evoprene (trademark) GC 5687|SEBS, Evoprene (trademark) GC 5688|SEBS, Evoprene (trademark) GC5689|SEBS, Evoprene(TM) 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(R) RG-38052 XRD1|TPE, megol(R) PUG 10|SEBS, megol(R) PUG 60|SEBS, megol(R) TA 60|SEBS, Monprene(registered trademark) RG-10130|TPE, Monprene(registered trademark) RG-10140|TPE, Monprene(registered trademark) RG-10150|TPE, Monprene(registered trademark) RG-10160|TPE, Monprene(registered trademark) RG-10170|TPE, Monprene(registered trademark) RG-15130|TPE, Monprene(registered trademark) RG-15140|TPE, Monprene(registered trademark) RG-15150|TPE, Monprene(registered trademark) RG-15160|TPE, Monprene(registered trademark) RG-15170|TPE, Monprene(registered trademark) RG-18240|TPE, Monprene(registered trademark) RG-18250|TPE, Mo Monprene(registered trademark) RG-18260|TPE, Monprene(registered trademark) RG-18270|TPE, Monprene(registered trademark) RG-19221 NAT|TPE, Monprene(registered trademark) RG-19255|TPE, Monprene(registered trademark) RG-20140|TPE, Monprene(registered trademark) RG-20160|TPE, Monprene(registered trademark) RG-20170|TPE, Monprene(registered trademark) RG-29068 NAT|TPE, Monprene(registered trademark) RG-29240 XRD1|TPE, RABALON(registered trademark) MJ4300C|TPE, RABALON(registered trademark) MJ5302C|TPE, RABALON(registered trademark) MJ6301C|TPE, RABALON(registered trademark) MJ7301C|TPE, RAYPRENE(registered trademark) NB221-S4050|TPE, RAYPRENE(registered trademark) NB221-S4051|TPE, RAYPRENE(registered trademark) NB221-S4052|TPE, RAYPRENE(registered trademark) NB221-S4053|TPE, tefabloc(registered trademark) TO 132|TPE, Telcar(registered trademark) TL-83-F943D22-NT BLU|TPE, THERMOLAST(registered trademark) K TF2CGT(Series:FC)|TPE, THERMOLAST(registered trademark) K TF3BTL (Series: FC / AP) | TPE, THERMOLAST® K TF3CGT (Series: FC) | TPE, THERMOLAST® K TF3STE (Series: FC / CS) | TPE, THERMOLAST® K TF4AAB (Series: FC / HE / tl) | TPE, THERMOLAST® K TF4BTL (Series: FC / AP) | TPE, THERMOLAST® K TF4CGT (Series: FC) | TPE, THERMOLAST® K TF4STE (Series: FC / CS) | TPE, THERMOLAST® K TF5AAC (Series: FC / HE / tl) | TPE, THERMOLAST® K TF5BTL (Series: FC / AP) | TPE, THERMOLAST® K TF5CGT (Series: FC) | TPE, THERMOLAST® KTF5STE (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, 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(trademark)GP; 2810-60N|TPE, Versaflex(trademark)GP; 2810-70N|TPE, Cawiton(registered trademark)MT920|SEBS; Cawiton(registered trademark)MT930|SEBS; Cawiton(registered trademark)MT940|SEBS; Cawiton(registered trademark)MT950|SEBS; Cawiton(registered trademark)MT960|SEBS; Cawiton(registered trademark)MT970|SEBS.

[0221] In carrying out the claimed invention, a person skilled in the art can understand and implement variations of the disclosed embodiments by examining the drawings, this disclosure, and the appended claims. In the claims, the words “equip,” “include,” and “have” do not exclude other elements or steps, and singular elements do not exclude plural elements.

[0222] The mere fact that certain techniques are described in different dependent claims does not indicate that combinations of these techniques cannot be used advantageously.

[0223] It should be noted that when the term "adapted to" is used in the claims or description, it is intended to be equivalent to the term "configured to".

[0224] No reference numeral in a claim should be construed as limiting its scope.

Claims

1. A method for manufacturing a biomaterial testing device, wherein the biomaterial testing device comprises a substrate on which at least one chamber unit is defined, and a support member for supporting a biomaterial received in the at least one chamber unit, the method comprising the step of using a mold to mold one of the substrate and the support member onto at least a portion of the other of the substrate and the support member, wherein at least a portion of the other of the substrate and the support member is positioned in or adjacent to the mold during the molding step.

2. The method according to claim 1, comprising the steps of: initially molding the base material or the support member in order to mold an initial molding component; and subsequently molding the unmolded of the base material and the support member onto the initial molding component.

3. The method according to claim 2, wherein the step of initial molding the substrate or the support member comprises heating the material to a temperature above room temperature so that the material becomes moldable in order to enable the formation of the initial molded component, and the step of molding the unmolded of the substrate and the support member onto the initial molded component is carried out before the initial molded component returns to room temperature, preferably while the temperature of the initial molded component is at least 80°C.

4. The method according to claim 2 or 3, comprising the steps of: placing at least a portion of the base member inside the mold or adjacent to the mold; initial molding the substrate or the support member on the base member; and removing the base member, on which the substrate and the support member have been molded, from the mold.

5. The method according to any one of claims 1 to 4, wherein the step of molding one of the substrate and the support member onto at least a portion of the other of the substrate and the support member comprises curing a precursor material, and optionally, curing comprises heating the precursor material.

6. The method according to any one of claims 1 to 5, further comprising the step of releasing the substrate together with the support member from the mold while the temperature of the substrate and the support member is higher than room temperature, preferably at least 40°C.

7. The method according to any one of claims 1 to 6, comprising the step of injection molding one of the base material and the support member onto at least a portion of the other of the base material and the support member.

8. The method according to any one of claims 1 to 7, comprising the step of adjusting the mold from a first configuration used for molding one of the base material and the support member to a second configuration used for molding the other of the base material and the support member.

9. The method according to any one of claims 1 to 8, wherein the support member is formed from a material that is more flexible and / or more pliable than the material forming the base material.

10. The method according to any one of claims 1 to 9, wherein the support member is formed from a material that is more biocompatible than the material forming the substrate, such that cells and / or tissues preferentially adhere to the support member.

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

12. The method according to any one of claims 1 to 11, wherein the support member comprises a polymer material bulk-modified in portions each containing polar groups, the polar groups of the portions are effective on the surface of the support member arranged to be in contact with the biomaterial received in the at least one chamber unit, and optionally the portions include fatty acid portions, and the polar groups include carboxylic acid groups of the fatty acid portions.

13. The method according to any one of claims 1 to 12, wherein the substrate is formed from a material having a glass transition temperature of at least 140°C, and / or the substrate is formed from a thermoplastic, preferably one or more selected from polycarbonate, polyetheretherketone, acrylonitrile butadiene styrene, polyetherimide, and polyethersulfone.

14. The method according to any one of claims 1 to 13, wherein the base material provides a side wall of the at least one chamber unit, a central portion of the support member provided within the at least one chamber unit, and a plurality of rib elements extending from the central portion and engaging with the corresponding side wall of the chamber unit.

15. The method according to any one of claims 1 to 14, wherein the support member comprises at least one structural feature that contacts the biomaterial received within the at least one chamber unit, and optionally, the at least one structural feature comprises a pair of flexible projections for supporting tissue on top of and between for each chamber unit.

16. The method according to any one of claims 1 to 15, wherein the at least one chamber unit is composed of a plurality of chamber units, and optionally the chamber units are arranged in one or more rows.

17. A mold system for molding a biological testing device using the method according to any one of claims 1 to 16, the mold system comprising a first mold component and a third mold component separable from the first mold component, wherein the first mold component and the third mold component are configured to combine to form a second mold configuration arranged to hold one of the substrate and the support member, the second mold configuration comprising the first mold component, the second mold component, and the one of the substrate and the support member, defining one or more openings in which the other of the substrate and the support member can be molded on the one of the substrate and the support member.

18. A substrate in which at least one chamber unit is defined, A support member for supporting a biomaterial received in at least one chamber unit, wherein one of the base material and the support member is molded on the other of the base material and the support member, and A biomaterial testing device equipped with the following features.

19. Use of the biomaterial inspection device according to claim 18 for inspecting biomaterials.

20. A step of providing a biomaterial in at least one chamber unit of a biomaterial testing device according to claim 18, wherein the step of providing the biomaterial in at least one chamber unit optionally comprises culturing cells in the at least one chamber unit, The steps include: exposing the aforementioned biological material to the drug to be tested; A drug testing method having the following characteristics.

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