Fluid device

Abstract

The fluid device 105 comprises an inlet 111, an outlet 112, and a first group of sub-chambers 121 including two or more first sub-chambers 121a, 121b, 121c, 121d arranged parallel to each other between the inlet 111 and the outlet 112. Each of the first sub-chambers 121a, 121b, 121c, 121d is configured to receive each corresponding cell culture insert 150. The fluid flow between the inlet 111 and the outlet 112 is evenly distributed and / or substantially identical among the first sub-chambers 121a, 121b, 121c, 121d.

Description

[Technical Field]

[0001] The present invention relates to a fluid apparatus and related methods for circulating substances through a branch chamber of the fluid apparatus. In particular, but not limited to, the present invention relates to a fluid apparatus capable of mimicking the circulatory system of an animal in drug research. [Background technology]

[0002] Drug research involves multiple stages. Generally, drug research begins with individual cell work and other experiments to assess the potential efficacy of a drug. These initial stages are usually performed in vitro. Promising candidates are then selected for subsequent in vivo animal experiments (usually in small rodents such as mice or rats). Overall, this approach results in a very low success rate for drug candidates between in vitro studies and in vivo animal experiments.

[0003] Improving the success rate of in vivo trials and reducing unnecessary in vivo trials is desirable for many reasons, including time, cost, and ethical considerations.

[0004] To bridge the gap between in vitro and in vivo drug research and to facilitate the elimination of unsuitable candidates that may be tested in vivo, specific devices exist that mimic drug distribution behavior in animals. Successful testing using such devices increases the likelihood that drug candidates will be successfully applied to human patients, as they will possess predictors of success in human in vivo testing derived from these devices.

[0005] One example of such a device is the HUMIMIC Chip4™ from TissUse Ltd. This device is used in drug research integrating up to four different organ models, such as the intestine, liver, kidney, and nerve tissue. The device is designed to investigate the toxic effects of drugs over time and to model processes such as the excretion and reabsorption of various substances in the kidney model using individual and complex microfluidic circuits. The complexity and size of the device stem from the fact that at least one sub-compartment is configured to accommodate a 96-well insert. Other sub-compartments are difficult to access for periodic sampling. The extremely small growth surface area of ​​the 96-well insert does not meet the volume requirements necessary for the analysis of intracellular drug concentrations using certain types of analytical techniques, such as high-performance liquid chromatography (HPLC) or liquid chromatography-mass spectrometry (LC-MS), making it difficult to normalize drug concentration relative to cell number.

[0006] Patent Document 1 (Li et al.) discloses a cell culture incubator device in which a culture plate is sandwiched between an upper plate and a bottom plate. In this configuration, fluid flow is possible by gravity / capillary action. None of the culture units are suitable for receiving cell culture inserts, and by design, sampling from any of the subcompartments is not possible. The flow is not evenly distributed among the subcompartments within each group of parallel-arranged subcompartments. This is because, at the very least, the central cells within each group of culture units are connected to both channels, while the outer culture units are connected to only one channel. Furthermore, the fluid is configured to flow into each culture unit, not to pass through each culture unit.

[0007] Patent document 2 (Ge et al.) discloses an organ chip integrated with a microelectrode array. In this configuration, fluid flow is enabled by gravity / capillary action. Although multiple cell culture wells are arranged in parallel, the fluid flow is not evenly distributed among the cell culture wells. This is because the two central wells are closer to the outlet channel than the two outer cell culture wells. Furthermore, none of the cell culture wells are suitable for accepting cell culture inserts, and because the wells are completely sealed, sampling from any of the sub-compartments is not possible by design. [Prior art documents] [Patent Documents]

[0008] [Patent Document 1] U.S. Patent Application Publication No. 2005 / 142656 [Patent Document 2] Chinese Patent No. 115109699 [Overview of the Initiative] [Problems that the invention aims to solve]

[0009] The object of the present invention is to solve and / or alleviate one or more problems related to the prior art.

[0010] The object of the present invention is to provide a fluid apparatus that mimics the drug distribution behavior in animals and enables sampling and analysis (e.g., HPLC or LC-MS analysis) of intracellular drug concentrations within its chamber. [Means for solving the problem]

[0011] According to a first aspect of the present invention, a fluid device is provided. The fluid device is The entrance and Exit and The system comprises a first group of sub-rooms, including two or more first sub-rooms arranged parallel to each other between the entrance and the exit. Each of the first sub-compartments is configured to receive each corresponding cell culture insert, The fluid flow between the inlet and the outlet is evenly distributed and / or is substantially the same among the first compartments.

[0012] The inlet can be in fluid communication with the first group of compartments.

[0013] The outlet can be in fluid communication with the first group of compartments.

[0014] The fluid device can be configured to enable fluid flow between the inlet and the outlet through each of the first compartments separately.

[0015] Generally, the inlet can be configured to receive fluid flow - for example, a biological fluid - into the fluid device. The outlet can be configured to allow the fluid to flow out of the fluid device.

[0016] The first compartments of the first group of compartments are arranged in parallel. In use, when fluid flows between the inlet and the outlet, there can be no fluid flow between the first compartments of the first group of compartments.

[0017] The device can be configured such that the flow rate of the fluid between the inlet and the outlet is substantially the same and / or is evenly distributed through each of the first compartments.

[0018] The fluid device can include a second group of compartments including one or more second compartments.

[0019] When the second group of compartments includes a plurality of second compartments - for example, two or more second compartments - the second compartments of the second group of compartments may be arranged in parallel between the inlet and the outlet. In use, when fluid flows between the inlet and the outlet, there can be no fluid flow between the second compartments of the second group of compartments.

[0020] The device can be configured such that the flow rate of the fluid between the inlet and the outlet is substantially the same through each of the second compartments.

[0021] The compartments of the second group may be provided between the inlet and the compartments of the first group.

[0022] The compartments of the second group may be provided between the compartments of the first group and the outlet.

[0023] The fluid device may include a third group of compartments including one or more third compartments arranged in parallel.

[0024] It is understood that the fluid device may further include any number of additional groups of compartments in parallel, as long as the fluid flow between the inlet and the outlet is substantially the same through each compartment of a given group of compartments.

[0025] The inlet may be in direct fluid communication with the group of compartments through each corresponding inlet fluid channel - for example, the two or more first compartments of the first group of compartments.

[0026] The outlet may be in direct fluid communication with the group of compartments through each corresponding outlet fluid channel, for example, the two or more first compartments of the first group of compartments.

[0027] Two or more groups of compartments may be provided.

[0028] A first group of compartments including two or more first compartments in direct fluid communication with the outlet through each corresponding outlet fluid channel may be provided. A second group of compartments including one or more second compartments in direct fluid communication with the inlet through each corresponding inlet fluid channel may be provided.

[0029] A first group of compartments including two or more first compartments in direct fluid communication with the inlet through each corresponding inlet fluid channel may be provided. A second group of compartments including one or more second compartments in direct fluid communication with the outlet through each corresponding outlet fluid channel may be provided.

[0030] Each sub-chamber of the second group may be in direct fluid communication with one or more first sub-chambers of the first group via their respective first fluid channels.

[0031] In one embodiment, the second group of subchambers may include one subchamber. This may be advantageous when the fluid device is intended to mimic the systemic blood flow of an animal, since the second subchamber can represent the heart of an animal. In such an embodiment, the inlet may be in direct fluid communication with the second subchamber via an inlet fluid channel.

[0032] The second subchamber can be in direct fluid communication with each of the first subchambers of the first group of subchambers via each corresponding first fluid channel.

[0033] Each of the first sub-chambers of the first group may be in direct fluid communication with the outlet via its corresponding outlet fluid channel.

[0034] In one embodiment, the second group of subchambers may include two or more second subchambers. In such an embodiment, the inlet may be in direct fluid communication with each of the second subchambers via each corresponding inlet fluid channel.

[0035] Each of the second subchambers may be in direct fluid communication with one or more of the first subchambers of the first group of subchambers via their respective first fluid channels.

[0036] Each of the first sub-chambers of the first group may be in direct fluid communication with the outlet via its corresponding outlet fluid channel.

[0037] The number of the first sub-units of the first group of sub-units may be less than, equal to, or greater than the number of the second sub-units of the second group of sub-units.

[0038] The inlet may be in direct fluid communication with the group of subchambers closest to the inlet via each corresponding inlet fluid channel—for example, the two or more first subchambers of the first group of subchambers or the one or more second subchambers of the second group of subchambers.

[0039] Each of the inlet fluid channels may be substantially the same in size and / or arranged symmetrically with respect to one another. This allows the fluid flow between the inlet and each of the sub-chambers of the group of sub-chambers closest to the inlet to be substantially the same and / or evenly distributed.

[0040] Each of the inlet fluid channels may be substantially the same in length, width, height, and / or diameter. If the cross-section of the inlet fluid channel is substantially circular or semicircular, the inlet fluid channels may be substantially the same in length and diameter.

[0041] The outlet may be in direct fluid communication with the group of subchambers closest to the outlet via each corresponding outlet fluid channel—for example, the two or more first subchambers of the first group of subchambers or the one or more second subchambers of the second group of subchambers.

[0042] Each of the outlet fluid channels may be substantially the same in size and / or arranged symmetrically with respect to one another. This allows the fluid flow between each of the subchambers in the group of subchambers closest to the outlet and the outlet to be substantially the same and / or evenly distributed.

[0043] Each of the outlet fluid channels may have substantially the same length, width, height, and / or diameter. If the outlet fluid channels are substantially circular or semicircular in cross-section, they may have substantially the same length and diameter.

[0044] A group of sub-chambers—for example, the two or more first sub-chambers of the first group of sub-chambers—may be in direct fluid communication with an adjacent group of sub-chambers—for example, the one or more second sub-chambers of the second group of sub-chambers—via each corresponding fluid connection channel—for example, the first fluid channel.

[0045] Each of the fluid connection channels between one group of subchambers and an adjacent group of subchambers may be substantially identical in size and / or arranged symmetrically with respect to one another. This makes it possible for the fluid flow between one group of subchambers and an adjacent group of subchambers to be substantially identical and / or evenly distributed.

[0046] Each fluid connection channel between one sub-chamber group and an adjacent sub-chamber group may have substantially the same length, width, height, and / or diameter. If the fluid connection channel is substantially circular or semicircular in cross-section, the fluid connection channels may have substantially the same length and diameter.

[0047] To the advantage of such a fluid apparatus configuration, the fluid flow between the inlet and the chambers of the group of chambers closest to the inlet is evenly distributed and / or substantially identical among the chambers of the group of chambers. If one or more further groups of chambers exist, the configuration of the fluid apparatus ensures that the fluid flow is evenly distributed and / or substantially identical among the chambers of each group of chambers. This ensures that the fluid flow rates between the chambers of each group of chambers are substantially equal. This facilitates the performance of experiments—e.g., drug experiments—intended to mimic the effects of a substance—e.g., a drug—in the cells of one or more organs represented by one or more chambers of the fluid apparatus.

[0048] Preferably, a group of sub-rooms or sub-rooms within each group of sub-rooms may be substantially identical in size and / or configuration.

[0049] The two or more first sub-compartments of the first group of sub-compartments may be substantially identical in size and / or configuration.

[0050] The one or more second sub-compartments of the second group of sub-compartments may be substantially identical in size and / or configuration.

[0051] The aforementioned one or more sub-units or sub-units of a further group may be substantially identical in size and / or configuration.

[0052] One or more sub-chambers of the fluid apparatus—for example, the sub-chambers—may define recesses or cavities configured to receive each corresponding cell culture insert.

[0053] One or more chambers of the fluid apparatus—for example, the aforementioned chambers—may be substantially spherical or hemispherical. One or more chambers of the fluid apparatus—for example, the aforementioned chambers—may be substantially cylindrical, cubic, or trapezoidal.

[0054] One or more sub-chambers of the fluid apparatus—for example, the sub-chambers—may be provided with connecting means for connecting to each corresponding cell culture insert. The connecting means may include any conventional connecting mechanism, such as one or more grooves configured to receive the engaging elements of each corresponding cell culture insert.

[0055] The fluid device may define a flow path between the inlet and the outlet.

[0056] The flow path may be defined by the channel and the subchamber. The flow path may be defined by the subchamber and the inlet fluid channel, the outlet fluid channel, and (if there are two or more subchamber groups) the fluid connection channels between subchambers of adjacent subchamber groups.

[0057] The fluid device may be manufactured from any material suitable for the flow of a fluid—for example, a biofluid or a pharmaceutical composition.

[0058] The fluid device may be manufactured from 3D printed material. The material may be a polymer material such as an acrylic material like VeroClear®. The flow channels may be formed by 3D printing the fluid device.

[0059] The fluid device may be manufactured from polymers, glass, silicon, or other suitable materials. The flow channels may be formed mechanically or chemically in the material of the fluid device, for example, by etching or machining.

[0060] Typically, the size of the fluid device may be in the range of approximately 6 cm x 8 cm.

[0061] Typically, the fluid channel may have a width and / or diameter in the range of about 1 to 5 mm, for example, about 2 mm.

[0062] The sub-chamber may have dimensions suitable for receiving cell culture inserts of corresponding sizes, such as a 96-well insert, a 48-well insert, a 24-well insert, a 12-well insert, or a 6-well insert, such as width, diameter, and / or depth. Preferably, the sub-chamber may have dimensions suitable for receiving cell culture inserts of corresponding sizes selected from a 24-well insert, a 12-well insert, or a 6-well insert, such as width, diameter, and / or depth. Such a configuration allows the apparatus to sample a sufficient volume of fluid to analyze intracellular drug concentrations using specific analytical techniques, such as high-performance liquid chromatography (HPLC) or liquid chromatography-mass spectrometry (LC-MS).

[0063] Typically, the subchamber may have a width and / or diameter in the range of approximately 14 to 37 mm—for example, approximately 16 to 35 mm, for example, approximately 20 to 25 mm, for example, approximately 22 mm. Generally, if the subchamber is configured to receive a 12-well insert, the subchamber may have a width and / or diameter in the range of approximately 20 to 25 mm—for example, approximately 22 mm. If the subchamber is configured to receive a 24-well insert, the subchamber may have a width and / or diameter in the range of approximately 14 to 18 mm, for example, approximately 16 mm. If the subchamber is configured to receive a 6-well insert, the subchamber may have a width and / or diameter in the range of approximately 33 to 37 mm, for example, approximately 35 mm.

[0064] The aforementioned sub-chamber may have a depth in the range of approximately 15-20 mm—for example, approximately 17.9 mm.

[0065] According to the second embodiment, a fluid device is provided. The fluid device is The entrance and Exit and A first group of sub-rooms including two or more first sub-rooms arranged parallel to each other between the entrance and the exit, A second group of sub-rooms, including one or more second sub-rooms arranged parallel to the entrance and the exit, Optionally comprising one or more sub-sub The inlet is in fluid communication with the first group of sub-chambers or the second group of sub-chambers via each of the (multiple) corresponding inlet channels. The outlet is in fluid communication with the second group of sub-chambers or the first group of sub-chambers via each of the (multiple) corresponding outlet channels. Each sub-compartment of a group is in fluid communication with one or more sub-compartments of an adjacent group via its corresponding fluid connection channel. Each sub-compartment is configured to receive its corresponding cell culture insert. Each of the inlet fluid channels is substantially the same in size and / or is arranged symmetrically to one another. Each of the outlet fluid channels is substantially the same in size and / or arranged symmetrically to one another. Each of the fluid connection channels between the sub-sub

[0066] To the advantage of this configuration, the fluid apparatus is configured to allow fluid flow between the inlet and the outlet through each subchamber, and the fluid flow may be evenly distributed and / or substantially identical among the subchambers of each group of subchambers.

[0067] In one embodiment, the fluid device is The entrance and Exit and A first group of subchambers including two or more first subchambers arranged in parallel (the outlets communicate directly with the two or more first subchambers of the first group of subchambers via their respective corresponding outlet fluid channels, and each of the inlet fluid channels is substantially the same in size and / or symmetrically arranged with respect to one another), A second group of sub-chambers including one or more second sub-chambers arranged in parallel (the inlets communicate directly with the one or more second sub-chambers of the second group of sub-chambers via each corresponding inlet fluid channel, and each of the outlet fluid channels is substantially the same in size and / or symmetrically arranged with respect to one another), The system may optionally include one or more sub-sub Each subcommitt of a group is in fluid communication—for example, direct fluid communication—with one or more subcommittees of an adjacent group via corresponding fluid connection channels, and each of the fluid connection channels between the subcommittees of an adjacent group is substantially the same in size and / or is arranged symmetrically with respect to one another. Each sub-compartment is configured to receive its corresponding cell culture insert.

[0068] The features described in relation to the first embodiment are also applicable to the apparatus of the second embodiment, and for the sake of brevity, they will not be repeated.

[0069] According to the third embodiment, a fluid system is provided. The fluid system comprises a fluid apparatus described in either the first or second embodiment, A pump that pressurizes and delivers fluid through the fluid device, It is equipped with.

[0070] The system may include an inlet conduit—for example, a cannula—configured to provide fluid communication between the pump and the inlet.

[0071] The system may include an outlet conduit—for example, a cannula—configured to provide fluid communication between the pump and the outlet.

[0072] The system may define a flow path for circulating fluid through the fluid device.

[0073] The pump may be configured to circulate the fluid from the inlet to the outlet through the fluid device. The pump may also be configured to circulate the fluid from the outlet to the inlet through the outlet conduit and the inlet conduit.

[0074] An advantage is that the pump may be configured to circulate the fluid from the inlet to the outlet through the fluid device, and further circulate it from the outlet to the inlet through the outlet conduit and the inlet conduit. The pump may be positioned between the outlet conduit and the inlet conduit.

[0075] The system may further include a fluid supply interface configured to allow a fluid—for example, a biofluid or a pharmaceutical composition—to be supplied into the system—for example, the channel.

[0076] The pump may be a peristaltic pump or a roller pump.

[0077] The system may comprise at least one—usually more—cell culture inserts. The cell culture inserts may be configured to be supplied to each corresponding branch chamber of the fluid apparatus.

[0078] The cell culture insert may include a 96-well insert, a 48-well insert, a 24-well insert, a 12-well insert, or a 6-well insert. An advantage is that the cell culture insert may include a 24-well insert, a 12-well insert, or a 6-well insert. Such a configuration allows the insert size to allow sampling of a sufficient volume of fluid for analysis by, for example, HPLC, LC-MS, or other analytical techniques. The insert size allows sampling of a fluid volume of approximately 10 μL or more—for example, at least 20 μL, at least 50 μL, or at least 75 μL. An advantage is that the cell culture insert may be sized to allow sampling of the insert without requiring the stopping of the fluid flow through the fluid apparatus. Typically, the cell culture insert includes a 12-well insert, or may be a 12-well insert. The 12-well insert has been found to be large enough to allow accurate and reproducible quantification from the insert by HPLC and / or LC-MS analysis, and small enough not to require culturing an excessive amount of cells to perform the experiment.

[0079] The cell culture insert may be seeded and / or filled with cells. The cells may represent a selected organ. One or more cell culture inserts may be seeded and / or filled with cells selected from cardiac cells, lung cells, hepatocytes, kidney cells, brain cells, pancreatic cells, spleen cells, etc.

[0080] In one embodiment, the second group of sub-chambers may include one sub-chamber configured to receive a cell culture insert into which cardiac cells are seeded or filled. This may be advantageous when the fluid device is intended to mimic the systemic blood flow of an animal, since the second sub-chamber can represent the heart of an animal. In such an embodiment, the inlet may be in fluid communication with the second sub-chamber via an inlet fluid channel.

[0081] The first chamber of the first group of chambers may be in fluid communication with the outlet via corresponding outlet fluid channels. Each of the first chambers may be configured to receive a cell culture insert into which different types of cells, selected from, for example, cardiac cells, lung cells, hepatocytes, kidney cells, brain cells, pancreatic cells, spleen cells, etc., are seeded or packed.

[0082] One or more cell culture inserts—for example, each cell culture insert—may have a semipermeable portion. Typically, the bottom of the cell culture insert may be configured to receive a semipermeable membrane. The semipermeable membrane may be configured to allow the passage of small molecules—for example, chemical substances such as drugs—but to prevent the passage of larger molecules or substances such as cells.

[0083] The inner surface of the cell culture insert may be filled with cells selected from, for example, cardiac cells, lung cells, hepatocytes, kidney cells, brain cells, pancreatic cells, spleen cells, etc. This allows a drug that has passed through the membrane into the insert to interact with the cells. This reaction can be evaluated by analyzing a sample taken from the insert.

[0084] The outer surface of the cell culture insert may be filled with cells, such as endothelial cells. This is advantageous because it allows for a more realistic simulation of the process by which a drug—for example, a drug contained in a fluid—passes through the capillary walls (represented here by endothelial cells on the outer surface of the cell culture insert) to reach an organ (represented here by cells on the inner surface of the insert).

[0085] According to the fourth aspect, a method is provided for testing the effect of a substance on one or more types of cells. The method is The step of providing the system described in the third aspect, The steps include providing a cell culture insert into one or more sub-compartments of the fluid apparatus, A step of supplying fluid to the flow path defined by the system and / or the fluid device of the system, The method includes a step of circulating the fluid so that it passes through the fluid apparatus and / or system.

[0086] The method may include the step of providing cell culture inserts to each sub-compartment of the fluid apparatus.

[0087] The method may include a step of sampling one or more of the cell culture inserts—for example, each cell culture insert.

[0088] According to the fifth aspect, a method for manufacturing a fluid apparatus according to the first or second aspect is provided. The method includes the step of manufacturing the fluid apparatus.

[0089] The method may include a step of providing an initial structure made of a base material, and a step of mechanically and / or chemically treating the base material, for example by etching or machining, to form sub-chambers and / or channels.

[0090] The method may include a step of manufacturing the fluid device by additive manufacturing—for example, 3D printing.

[0091] The examples relating to this disclosure may be formed using additive manufacturing processes. A common example of additive manufacturing is 3D printing, but other additive manufacturing methods are also available. Rapid prototyping or rapid manufacturing may also be used as terms to describe additive manufacturing processes.

[0092] In this specification, “additive manufacturing” generally refers to a manufacturing process in which a three-dimensional part is “laminated” or “additively manufactured” by providing a series of continuous material layers to each other. This is in contrast to certain removal processes (e.g., milling or drilling) that sequentially remove material to manufacture the part. The continuous material layers can generally be integrated to form a single part that can have various integrated components. In particular, this manufacturing process can integrally form the examples of this disclosure and include various features that were not possible with conventional manufacturing methods.

[0093] The additive manufacturing method described in this application enables the production of parts of any size and shape with various characteristics that were previously thought impossible with conventional manufacturing methods. Additive manufacturing can create complex shapes without using tools, molds, jigs, etc., and with little to no waste material. Instead of machining parts from blocks of plastic or metal, much of which is cut and discarded, only the materials used in additive manufacturing are required to form the parts.

[0094] Appropriate additive manufacturing technologies relating to this disclosure include, for example, fused deposition modeling (FDM), selective laser sintering (SLS), inkjet and laser jet 3D printing, stereolithography (SLA), direct selective laser sintering (DSLS), electron beam sintering (EBS), electron beam melting (EBM), laser engineered net shaping (LENS), electron beam additive manufacturing (EBAM), laser net shaping manufacturing (LNSM), direct metal deposition (DMD), digital light processing (DLP), continuous digital light processing (CDLP), direct selective laser melting (DSLM), selective laser melting (SLM), direct metal laser melting (DMLM), direct metal laser sintering (DMLS), material jetting (MJ), nanoparticle jetting (NPJ), drop-on-demand (DOD), binder jetting (BJ), multi-jet fusion (MJF), additive manufacturing (LOM), and other known processes.

[0095] The additive manufacturing process described herein may be used to form parts using any suitable material. For example, the material may be plastic, metal, composite material, concrete, ceramic, polymer, epoxy, photopolymer resin, or any form or combination thereof such as solid, liquid, powder, sheet material, wire, etc. More specifically, according to the exemplary embodiments of the subject matter herein, the additive manufactured parts described herein may be formed in part, whole, or in combination of materials, including, but not limited to, pure metals, nickel alloys, chromium alloys, titanium, titanium alloys, magnesium, magnesium alloys, aluminum, aluminum alloys, iron, iron alloys, stainless steel, and nickel or cobalt-based superalloys (e.g., Inconel®, a registered trademark of Special Metals Corporation). These materials are examples of materials suitable for the additive manufacturing process described herein and are suitable for manufacturing the examples described herein.

[0096] Typically, the additive manufacturing processes described herein may use polymer materials, such as acrylic materials like VeroClear®.

[0097] As described above, the additive manufacturing process disclosed herein makes it possible to form a single part from multiple materials. Therefore, the examples described herein may be formed from any mixture of the above materials. For example, a part may include multiple layers, segments, or parts formed by different materials, processes, and / or different additive manufacturing equipment. In this way, it is possible to construct parts with different materials and material properties to meet the requirements of a particular application. Furthermore, although all parts described herein are constructed by additive manufacturing processes, it should be noted that in other embodiments, all or part of these parts may be formed by casting, machining, and / or other suitable manufacturing processes. In fact, any suitable combination of materials and manufacturing methods may be used to form these parts.

[0098] Additive manufacturing processes typically produce parts based on three-dimensional (3D) information about those parts, such as a three-dimensional computer model (or design file) of the part.

[0099] Therefore, the examples described herein include not only the products or components described herein, but also methods for manufacturing such products or components by additive manufacturing, and computer software, firmware, or hardware for controlling the manufacturing of such products by additive manufacturing.

[0100] The structure of one or more parts of the aforementioned product may be represented digitally in the form of a design file. A design file or computer-aided design (CAD) file is a configuration file that encodes one or more surface or volumetric configurations of the shape of the product. In other words, the design file represents the geometric arrangement or shape of the product.

[0101] The design file can take any known or future file format. For example, the design file may be in the stereolithography or "Standard Tessellation Language" (.stl) format created for 3D Systems' stereolithography CAD program, or in the American Society of Mechanical Engineers (ASME) standard Additive Manufacturing File (.amf) format (an extensible markup language (XML)-based format designed to allow any CAD software to describe the shape and composition of any three-dimensional object produced by any additive manufacturing printer).

[0102] Furthermore, examples of design file formats include AutoCAD (.dwg) files, Blender (.blend) files, Parasolid (.x_t) files, 3D Manufacturing Format (.3mf) files, Autodesk (3ds) files, Collada (.dae) files, Wavefront (.obj) files, and many other file formats exist.

[0103] Design files may be created using modeling software (e.g., CAD modeling), or they may be created by scanning the surface of the product and measuring its surface structure.

[0104] Once the design file is obtained, it may be converted into a set of computer-executable instructions that, when executed by the processing apparatus, control an additive manufacturing apparatus to produce a product according to the geometric arrangement specified in the design file. This conversion may also convert the design file into slices or layers sequentially formed by the additive manufacturing apparatus. The instructions (so-called geometric code or "G-code") may be calibrated to a particular additive manufacturing apparatus and may specify the precise location and amount of material to be formed at each stage of the manufacturing process. As described above, the formation may be carried out by deposition, sintering, or any other additive manufacturing method.

[0105] The code or instruction may be translated between different formats, converted into a set of data signals and transmitted, received as a set of data signals, converted into code, and stored as necessary. The instruction may be an input to the additive manufacturing system and may be provided by a part designer, intellectual property (IP) provider, design company, operator or owner of the additive manufacturing system, or other supplier. The additive manufacturing system may use any of the techniques or methods disclosed herein to execute the instruction and manufacture the product.

[0106] Design files or computer-executable instructions may be stored in a (temporary or non-temporary) computer-readable storage medium (e.g., memory, storage system, etc.) that stores code or computer-readable instructions representing the product to be manufactured. As described above, the code or computer-readable instructions define the product that, when executed by an additive manufacturing system, can be used to physically produce the object. For example, the instructions may include a strictly defined 3D model of the product and may be generated from a well-known computer-aided design (CAD) software system such as AutoCAD®, TurboCAD®, or DesignCAD 3D Max. Alternatively, three-dimensional information of the part may be obtained by scanning a model or prototype of the part.

[0107] Therefore, by controlling the additive manufacturing apparatus according to the computer executable instructions, the additive manufacturing apparatus may be instructed to output one or more parts of the product. These may be printed in assembled or unassembled form. For example, different sections of the product may be printed individually (as a kit of unassembled parts) and then assembled. Alternatively, various parts may be printed in assembled form.

[0108] Based on the above, the embodiments include a manufacturing method by additive manufacturing. This includes a step of obtaining a design file representing the product and instructing an additive manufacturing apparatus to manufacture the fluid device according to the design file. The additive manufacturing apparatus may include a processing unit configured to automatically convert the design file into computer-executable instructions and control the manufacturing of the fluid device. In these embodiments, the fluid device can be manufactured automatically once the design file itself is input to the additive manufacturing apparatus. Therefore, in this embodiment, the design file itself may be considered as a computer-executable instruction that causes the manufacturing of the fluid device. Alternatively, the design file may be converted into instructions by an external computer system, and the resulting computer-executable instructions may be provided to the additive manufacturing apparatus.

[0109] In view of the foregoing, the design and manufacture of the embodiments covered by this specification and the operations described herein may be implemented using digital electronic circuits, or computer software, firmware, hardware (including structures disclosed herein and their structural equivalents), or one or more combinations thereof. For example, hardware includes processors, microprocessors, electronic circuits, electronic components, integrated circuits, etc. The embodiments covered by this specification may also be implemented using one or more computer programs, i.e., computer program instructions of one or more modules encoded on a computer storage medium, which may be executed by a data processing device or for controlling its operation. In addition or alternatively, the program instructions may be encoded on an artificially generated propagating signal. The propagating signal is, for example, a machine-generated electrical, optical, or electromagnetic signal that is produced to be transmitted to a suitable receiving device to encode information for execution by a data processing device. The computer storage medium may be a computer-readable storage device, a computer-readable storage board, a random or serial access memory array or device, or one or more combinations thereof. Furthermore, the computer storage medium may not be a propagating signal itself, but may be the source or destination of computer program instructions encoded on an artificially generated propagating signal. The computer storage medium may be one or more individual physical components or media (for example, multiple CDs, disks, or other storage devices).

[0110] In this specification, additive manufacturing techniques are described as enabling the production of complex objects by constructing them point by point, layer by layer, in a vertical direction; however, other manufacturing methods are also possible and are within the scope of this specification. For example, while this specification refers to the formation of continuous layers by laminating materials, those skilled in the art will understand that the methods and structures disclosed herein can be implemented using any additive manufacturing technique or other manufacturing technique.

[0111] Features described in relation to any aspect of the present invention are equally applicable to any other aspect and will not be repeated for brevity. For example, features described in relation to an apparatus are also applicable to a method, and vice versa. [Brief explanation of the drawing]

[0112] Embodiments of the present invention will be described with reference to the accompanying drawings.

[0113] [Figure 1] This figure shows a fluid apparatus according to the first embodiment. [Figure 2] This figure shows the fluid apparatus shown in Figure 1 with a cell culture insert attached. [Figure 3] This figure shows cell culture inserts for use with the apparatus shown in Figure 1. [Figure 4] This figure shows a fluid system according to the second embodiment. [Figure 5] This figure shows a visual investigation of the flow using an aqueous food coloring medium with the apparatus shown in Figure 1. [Figure 6] This figure shows the maximum intensity projection image obtained by PET / CT scan when tested using the first apparatus shown in Figure 1 at a flow rate of 1.5 mL / min. [Figure 7] This figure shows another fluid apparatus as an example. [Figure 8] This figure shows the maximum intensity projection image obtained by PET / CT scan when tested using the apparatus shown in Figure 7 at a flow rate of 1.5 mL / min. [Figure 9] This is a schematic diagram of the layout of the sub-units of the apparatus shown in Figure 1, which is equipped with cell inserts filled with cells. [Figure 10] Figure 6 shows the time-time radioactivity curve obtained from testing the apparatus shown in Figure 1. [Figure 11] This chromatogram shows inter-laboratory crosstalk, indicated by measurable drug uptake and the presence of hepatocyte metabolites in other sub-laborises. [Figure 12] This chromatogram shows inter-laboratory crosstalk, indicated by measurable drug uptake and the presence of hepatocyte metabolites in other sub-laborises. [Figure 13] This chromatogram shows inter-laboratory crosstalk, indicated by measurable drug uptake and the presence of hepatocyte metabolites in other sub-laborises. [Figure 14] This chromatogram shows inter-laboratory crosstalk, indicated by measurable drug uptake and the presence of hepatocyte metabolites in other sub-laborises. [Figure 15] Figure 4 shows a linear regression of Ki derived from intracellular [18F]FDG measured within the system insert, plotted against human vivo with SUVmean, r²=0.7966, p=0.0416, and n=1 in Pearson correlation. [Modes for carrying out the invention]

[0114] In this disclosure, unless otherwise indicated by context, several terms have the meanings set forth below.

[0115] "To include," "to contain," "to have," or variations thereof are understood here to mean the inclusion of a specific element, integer, or process, or a group of elements, integers, or processes, and not the exclusion of any other element, integer, or process, or a group of elements, integers, or processes.

[0116] "Consists of," "composed of," or variations thereof are understood to mean the inclusion of a particular element, integer, or process, or group of elements, integers, or processes, and the exclusion of any other element, integer, or process, or group of elements, integers, or processes.

[0117] In this specification, the term "about" is used to modify a number or value, referring to a value within ±5% of the specified value. For example, if the temperature is specified as about 5 to about 13°C, it includes temperatures from 4.75 to 13.65°C.

[0118] When referring to the physical state of a substance (e.g., liquid or solid), unless otherwise indicated by the context, it refers to the state of the substance at 25°C and atmospheric pressure.

[0119] In this specification, the term "direct fluid communication" means that there are no other sub-compartments or groups of sub-compartments between the indicated parts.

[0120] As described above, the inventors have discovered that they can provide a fluid apparatus that enables uniform fluid flow between the inlet and outlet through each sub-chamber of the same group.

[0121] Figure 1 shows a fluid apparatus 105 according to the first embodiment.

[0122] In this embodiment, the apparatus 105 has a body 110 3D printed from an acrylic polymer, namely VeroClear®. However, it will be understood that the fluid apparatus 105 can also be manufactured by other manufacturing techniques, such as chemical etching or machining.

[0123] The fluid device 105 includes an inlet 111 configured to receive a fluid, such as a biological fluid, into the fluid device, and an outlet 112 configured to discharge the fluid from the fluid device.

[0124] In this embodiment, the fluid apparatus comprises two sub-chamber groups.

[0125] The first group of sub-compartments 121, located closest to the exit 112, comprises four sub-compartments 121a, 121b, 121c, and 121d arranged in parallel.

[0126] The second group of sub-rooms 122, located closest to the entrance 111, has a single second sub-room 122.

[0127] Inlet 111 is in direct fluid communication with the second sub-chamber 122 via inlet channel 131.

[0128] Outlet 112 is in direct fluid communication with sub-chambers 121a, 121b, 121c, and 121d of the first group 121 via outlet channel 132.

[0129] Advantageously, the fluid device 105 is configured to allow fluid flow between the inlet 111 and the outlet 112, through each of the first branch chambers 121a, 121b, 121c, and 121d individually, and the fluid flow is distributed evenly or substantially identically among the first branch chambers 121a, 121b, 121c, and 121d.

[0130] The second sub-chamber 122 is in direct fluid communication with each of the first sub-chambers 121a, 121b, 121c, and 121d via their respective fluid connection channels 133a, 133b, 133c, and 133d.

[0131] Each of the fluid connection channels 133a, 133b, 133c, and 133d between the second branch chamber 122 and the first branch chambers 121a, 121b, 121c, and 121d is substantially identical in dimensions and arranged symmetrically with respect to one another. In this embodiment, they are arranged at a fixed angle of about 60°. Each of the fluid connection channels 133a, 133b, 133c, and 133d is substantially identical in length of about 5.1 mm and in diameter of about 2 mm.

[0132] Advantageously, this configuration makes it possible for the fluid flow between the second subchamber 122 and the first subchambers 121a, 121b, 121c, and 121d of the first group of subchambers 121 to be substantially identical and / or evenly distributed.

[0133] Outlet 112 is in direct fluid communication with each of the sub-chambers 121a, 121b, 121c, and 121d of the first group 121 via their respective corresponding outlet fluid channels 134a, 134b, 134c, and 134d. Each of the outlet fluid channels 134a, 134b, 134c, and 134d is substantially identical in dimensions and is arranged symmetrically with respect to one another.

[0134] Each outlet fluid channel 134a, 134b, 134c, and 134d is substantially identical, with a length of approximately 12.4 mm and a diameter of approximately 2 mm.

[0135] Advantageously, this configuration allows the fluid outflow from each of the first sub-chambers 121a, 121b, 121c, and 121d to be substantially identical and / or evenly distributed, thus avoiding bias in the minimum resistance path among the same group of sub-chambers, i.e., the first sub-chambers 121a, 121b, 121c, and 121d.

[0136] In this embodiment, the first and second outlet fluid channels 134a and 134b merge into a first auxiliary channel 135a that is in direct fluid communication with outlet 132. Similarly, the third and fourth outlet fluid channels 134c and 134d merge into a second auxiliary channel 135b that is in direct fluid communication with outlet 132.

[0137] The first and second auxiliary channels 135a and 135b are arranged substantially symmetrically with respect to the outlet, have substantially the same length, and are substantially the same in diameter, approximately 2 mm. This configuration also equalizes the flow of fluid passing through and out of each of the first subchambers 121a, 121b, 121c, and 121d, and avoids bias in the minimum resistance path among the same group of subchambers, i.e., the first subchambers 121a, 121b, 121c, and 121d.

[0138] Preferably, the sub-rooms within each sub-room group are substantially identical in size and / or configuration.

[0139] In this embodiment, the first sub-chambers 121a, 121b, 121c, and 121d of the first group of sub-chambers 121 are substantially identical in size and configuration. The first sub-chambers 121a, 121b, 121c, and 121d are substantially cylindrical, with a diameter of approximately 22 mm and a depth of approximately 17.9 mm. This configuration promotes uniform fluid flow through each of the first sub-chambers 121a, 121b, 121c, and 121d of the first group of sub-chambers 121.

[0140] For convenience, in this embodiment, the second sub-chamber 122 is also substantially cylindrical, with a diameter of approximately 22 mm and a depth of approximately 17.9 mm.

[0141] For convenience, the size of the sub-compartment was selected to accommodate a 12-well cell culture insert, such as the 12-well insert 150 shown in Figure 3. The 12-well insert was found to be large enough to allow accurate and reproducible quantification from the insert by HPLC and / or LC-MS analysis, and small enough not to require culturing an excessive amount of cells for the experiment. While not intended to be theoretically constrained, if the insert size is too small, for example, a 96-well insert, the cell count is likely too low to obtain a measurable concentration of the test compound / metabolite, falling below the detection limit of analytical techniques such as HPLC and LC-MS.

[0142] Each subcompartment has connecting means in the form of recesses 125 arranged to receive complementary engaging members 151 of each corresponding cell culture insert 150. It will be understood that the specific shape and size of the recesses 125 may be configured to fit or receive complementary engaging members 151 of each cell culture insert 150 intended for use with the device 105. In some embodiments, multiple sets of recesses 125 may be provided, each recess 125 configured to fit or receive complementary engaging members 151 of a particular type of cell culture insert 150 intended for use with the device 105. This may increase the versatility of the device 105.

[0143] Referring to Figure 3, the insert 150 has a main body 154 that defines a cavity 155 inside. The insert 150 also has a flange portion 152 configured to engage with the edge of each chamber. The flange portion 152 allows the bottom portion 153 of the insert 150 to be positioned at a predetermined depth or height, thereby allowing the bottom portion 153, which usually has a semipermeable membrane, to be placed within the flow path of the fluid device during use. This allows one or more substances in the fluid to permeate through the bottom portion 153, for example, a semipermeable membrane, into the cavity 155 during use. It will be understood that the semipermeable membrane may have different pore sizes depending on the desired movement between the fluid and the chamber cavities 155. For example, a pore size of 3 μm allows the passage of fluid and substrate, larger pores (e.g., 12 μm) allow intercellular interactions when the aim is to make the “organ” chambers more complex and closer to those in vivo, and smaller pores (e.g., 0.22 μm) are used when the aim is to prevent interactions between the two layers.

[0144] Figure 2 shows the fluid apparatus 105 of Figure 1, with cell culture inserts 150 placed in each of the subcompartments 121a, 121b, 121c, 121d, and 122, and the engaging members 151 of the cell culture inserts 150 engaged with the corresponding recesses 125 of the subcompartments 121a, 121b, 121c, 121d, and 122.

[0145] Figure 4 shows a fluid system 260 according to one embodiment. The system 260 includes the fluid device 105 shown in Figure 1. The system 260 includes a pump, which in this embodiment is a peristaltic pump or a roller pump 270. The pump 270 includes a cassette 274 that generates pressure between cannulas 271 connected to the pump to induce flow. In this embodiment, the pump has two cassettes, but it may also have, for example, four cassettes, and thus multiple fluid devices 105 can be operated by the pump 270 simultaneously.

[0146] The pump 270—in this embodiment, the cassette 274—is connected to the inlet 111 via an inlet cannula 271 configured to provide fluid communication between the pump 270 and the inlet 111, and is also connected to the outlet 112 via an outlet cannula configured to provide fluid communication between the pump 270 and the outlet 112.

[0147] Therefore, system 260 defines a flow path for circulating fluid through the fluid device 105. Pump 270 is configured to circulate the fluid from the inlet 111 through the fluid device 105 to the outlet 112, and further from the outlet 112 back to the inlet 111 via the outlet cannula 272, pump 270, and inlet cannula 271. This configuration allows for continuous circulation of the fluid between the inlet 111 and outlet 122 through the fluid device 105, and further between the outlet 112 and inlet 111 via pump 270, thereby mimicking the whole-body circulation of mammals.

[0148] The system further includes a fluid supply interface 273 connected to the flow path, which communicates with an outlet cannula 272, allowing a fluid, such as a biofluid, to be supplied to the flow path. In this embodiment, a syringe 275 containing fluid is connected to the fluid supply interface 273. The apparatus shown in this embodiment can mimic medical intravenous injection into a venous blood pool. It will be understood that other supply interface devices are also conceivable depending on the specific research being conducted. For example, in other embodiments, a separate inlet to a "bowel" branch / insert can mimic drug absorption by oral ingestion rather than injection.

[0149] Although not explicitly shown in Figure 4, it will be understood that when in use, the cell culture insert 150 is placed in each subcommittee 121a, 121b, 121c, 121d, and 122. Advantageously, the cell culture insert is a 12-well insert. The 12-well insert was found to be large enough to sample a suitable amount of fluid from the insert for HPLC analysis, and small enough not to require culturing an excessive amount of cells to perform the experiment. This allows for sampling of approximately 10 μL or more of fluid from the insert 150, e.g., at least 20 μL, e.g., at least 50 μL, e.g., at least 75 μL. Advantageously, the cell culture insert 150 is sized to allow sampling of the insert 150 without interrupting the fluid flow through the fluid apparatus 105 during use.

[0150] [Optimization of fluid systems] [Basic Visual Testing] To test for defects in the 3D-printed fluid apparatus, a basic visual inspection of the fluid flow using the fluid apparatus was performed with an aqueous food coloring medium 380. An early version of one embodiment of the apparatus 305 shown in Figure 5 was performed using the apparatus in Figure 4. The apparatus 305 in Figure 5 is generally similar to the apparatus 105 in Figure 1, with corresponding components numbered by adding "200". Figure 5 shows that the fluid is not passing through region 382 of the first auxiliary channel 335a, indicating the presence of a blockage. This simple method allows the various channels of the fluid apparatus 305 to be tested before being used in full-scale research, such as drug research.

[0151] [Optimization of body-on-chip devices] The perfusion performance of the fluid device 105 was tested in combination with a preclinical PET / CT small animal scanner (nanoPET / CT, Mediso) using [18F]FDG or [18F]NaF supplied by Queen's Medical Research Imaging (QMRI) Edinburgh Imaging Facility (University of Edinburgh, UK). This was performed by visually inspecting the distribution of [18F] tracers using PMOD image analysis software (PMOD Technologies). PET scans were acquired in 1:5 coincidence mode. Subsequently, CT scans were acquired (semicircular orbit, maximum field of view, 480 projection, 35kVp, 400ms, 1:4 binning) with structural overlay and attenuation correction. Tracers were circulated to the fluid device using an MS-4 / 12 Reglo digital pump (ISMATECTM), 270. Once perfusion was deemed uniform, regions of interest were plotted using PMOD to extract time-radioactivity curves, input functions, and venous output for each sub-chamber.

[0152] Figure 6 shows the maximum intensity projection image of a PET / CT scan performed using the first apparatus shown in Figure 1 at a flow rate of 1.5 mL / min.

[0153] As described above, the flow was measured using [18F]FDG / [18F]NaF and the NanoScan MicroPET / CT system, and the gold standard flow measurements obtained from PET were superimposed with structural images obtained from CT scans. As shown in Figure 6, the resulting average images and time-time radioactivity curves indicate that radioactivity is evenly distributed throughout the organ chambers and within the connecting capillaries.

[0154] [Examples of alternative designs based on comparative examples] Figure 7 shows an alternative fluid apparatus 405 based on a comparative example. Apparatus 405 in Figure 7 is generally similar to apparatus 105 in Figure 1, with corresponding components numbered by adding "300". However, in Figure 7, the fluid connection channels 433a, 433b, 433c, and 433d between the second branch chamber 422 and the first branch chambers 421a, 421b, 421c, and 421d are not substantially identical in dimensions and are not symmetrically arranged with respect to each other. Furthermore, the outlet channels 432 are not substantially identical in shape and dimensions and are not symmetrically arranged with respect to each other. As a result, it was found that the fluid flow between the second branch chamber 422 and each of the first branch chambers 421a, 421b, 421c, and 421d of the first group of branch chambers 421 was neither identical nor evenly distributed. This is supported by the maximum intensity projection images of PET / CT scans performed using the apparatus in Figure 7 at a flow rate of 1.5 mL / min, as shown in Figure 8.

[0155] [Examples and Experiments] [Double seeding of endothelial cells and "organ" cells onto the opposite side of a cell culture insert] Three Corning 3μm polyethylene terephthalate 12-well inserts (insert 150 in Figure 3) were placed upside down in each well of a 6-well plate, and 1 mL of Dulbecco phosphate-buffered saline (DBPS) was added to each well to prevent evaporation. Human umbilical vein endothelial cells (HUVECs) (PromoCell®, Germany) were placed at a density of 25,000 cells / cm² on the underside of the transwell inserts. 2The cells were seeded at a density of 1 mL in total volume and left for 4 hours to adhere to the membrane. After 4 hours, the inserts were returned to their original orientation and carefully transferred from the DBPS plate to a plate containing fresh endothelial cell proliferation medium, and left overnight for equilibration and division. The following day, organ-specific cell types were seeded into the inserts at their respective optimal seeding densities and left for 24 hours to adhere to the membrane. The three cell types used were human dermal fibroblasts (HDFibro), which produce and deposit extracellular matrix components and are expected to have quantitative drug uptake; HepG2 (ECCAC) liver cancer cell line, which shows metabolite production; and SH-SY5Y (ECCAC) neuroblast cell line, which has little drug uptake but may have metabolite uptake if there is inter-sector crosstalk. On the day of the study, each insert was observed individually, and it was confirmed that endothelial cells were present on the outer surface of the bottom 153 of the insert, and organ cells were present on the inner surface.

[0156] [Body-on-a-chip research using living cells] The tip was filled with 11 mL of endothelial cell medium, and 2.4 mM docetaxel was prepared with 500 μL of endothelial medium and dimethyl sulfoxide (76:24, v:v). 22.5 MBq of [18F]FDG was added to the docetaxel administration solution to a final volume of 1 mL, so that the final circulating concentration upon injection into the tip was 100 μM docetaxel in 1% (v:v) DMSO. Next, as shown in Figure 9, five 12-well inserts were placed in the tip, the inlet and outlet were connected with cannulas, and the tip was placed on the NanoScan PET / CT bed.

[0157] Figure 9 is a schematic diagram of the sub-compartmental arrangement of the apparatus in Figure 1 where cell inserts are placed. IO stands for "inserts only," and the two sub-compartments 121a and 122 require primary cells.

[0158] The peristaltic pump was set to a flow rate of 1.5 mL / min to initiate flow to the tip. The radioactive tracer and drug were injected from inlet 111, and PET acquisition was performed for 60 minutes in 1:5 scan mode to confirm uniform flow distribution through sub-compartments 122, 121a, 121b, 121c, and 121d. Immediately after the scan was completed, CT acquisition (semicircular orbit, maximum field of view, 480 projection, 35 kVp, 400 ms, 1:4 binning) was immediately started to perform structural overlay and attenuation correction of the PET data. After completion, the tip was removed from the PET / CT and placed behind lead. The insert was removed and placed in each well of a 6-well plate, the culture medium in the insert was removed, and the cells were washed with DPBS. The cells were detached with 200 μL of Tryple (ThermoFisher Scientific), of which 20 μL was used for cell counting, and the remainder was transferred to a 500 μL Eppendorf® tube. The Eppendorf® tube was rotated at 1000 × g for 5 minutes to generate centrifugal force to form a pellet, and the supernatant was removed. The cell pellet was resuspended in 75 μL of dH2O to prevent aggregation, and then dissolved with 150 μL of acetonitrile. Another 75 μL of dH2O was added to bring the final solution to a 50:50 (v:v) dH2O:acetonitrile ratio, matching the mobile phase and docetaxel standard for HPLC analysis. The Eppendorf® tube was rotated again to generate centrifugal force, and the supernatant was collected for HPLC analysis of intracellular drug concentrations.

[0159] Figure 10 shows the time-time radioactivity curve obtained from testing the apparatus shown in Figure 1, which was described in Figure 6.

[0160] The input function points to the inlet channel 131 leading to cardiac branch 122 and fluctuates due to the pulsating flow of the peristaltic pump 270. Cardiac branch 122 shows a peak immediately afterward, and then the input function and cardiac radioactivity decrease as the radioactive tracer is distributed from the heart to other organs (branchs 121a, 121b, 121c, 121d), reaching equilibrium in about 4 minutes.

[0161] Figures 11-14 are chromatograms showing measurable drug uptake and inter-laboratory crosstalk, indicated by the presence of hepatocyte metabolites in other sub-laboriae. -Figure 11: Chromatogram of 0.1 mg / mL docetaxel injection in mobile phase, retention time 7.473 minutes. -Figure 12: Chromatogram of lysed hepatocytes injected 1 hour after docetaxel injection into the novel invention, showing the presence of the same metabolite. -Figure 13: Chromatogram of lysed SH-SY5Y cells injected in the brain sub-unit. As expected, there is no drug uptake, but it shows the presence of metabolites generated in the liver sub-unit. -Figure 14: Chromatogram of injected lysed human dermal fibroblasts, showing measurable drug uptake and the presence of metabolites.

[0162] [DDChip [18F]FDG Dynamic Testing Method] HUVEC barrier inserts were prepared (see above), and each organ cell was seeded 24 hours after being transferred to a 12-well plate with HUVEC (PromoCell®, Germany) attached. The cells were cultured for 48 hours before the start of the experiment to allow the organ cells to adhere and proliferate. HUVEC was differentiated at passages 4-5, SH-SY5Y (ECCAC, 94030304) at passages 2-3, SA7K (Sigma™, MA, USA) and HepG2 (ECACC, 85011430) at passages 5-6, and HCM and HBEPC (PromoCell, Germany) at passage 2. Apparatus 105 (see Figure 4) was sterilized in a sterile hood in 70% isopropanol for 30 minutes, dried, and stored in a sterile container. During the test, device 105 was connected to an MS-4 / 12 Reglo digital pump 270 (ISMATECTM, Wertheim, Germany) set to 1.5 mL / min, and simulated intravenous injection of docetaxel or [18F]FDG using a closed circuit with a three-way inlet (see Figure 4).

[0163] The device was filled with 11 mL of endothelial cell growth medium preheated to 37°C, and the total volume was made 12 mL with a 1 mL injection. Insert 150 representing the heart, lungs, liver, kidneys, and brain with an endothelial barrier was placed in device 105 and placed in an incubator to prepare for [18F]FDG injection. [18F]FDG was prepared in physiological saline, with a total volume of 1 mL in the endothelial medium, a concentration of 50.04 ± 11.96 MBq / mL (mean ± SD, n = 9), and a circulating concentration in device 105 of 4.17 ± 1.00 MBq / mL. After starting the flow and injecting the [18F]FDG dosing solution into system 260, inlet 111 was tightened to prevent backflow. At each time point, cassette 274 was removed from system 260 to stop the flow, insert 150 was recovered without affecting other devices, and the intracellular [18F]FDG concentration was measured for kinetic modeling.

[0164] [DDChip [18F]FDG Kinetic Test Results] Cell lysates had intracellular [18F]FDG that was measurable from a minimum of 1 minute in all compartments. Ki (influx rate of the irreversible binding model) derived from the Patlak model was highest in the brain compartment at 4.73×10 -3 mL / cm 3 / min, followed by the kidneys, heart, liver, and lungs in that order at 2.84×10 -3 , 2.43×10 -3 , 1.01×10 -3 , 1.93×10 -4 mL / cm 3 / min. These values were compared with the in vivo SUVmean values (SUVmean is the mean standardized uptake value obtained by normalizing the tissue concentration by the administered dose and body weight) in healthy human volunteers in each organ compartment and are shown in Figure 15. The derived K i showed a significant correlation (r 2 = 0.7966, p = 0.0416, n = 1, Pearson correlation) with the in vivo SUVmean. A significant linear correlation was observed between the kinetics in this system 260 and human [18F]FDG kinetics, indicating that this system 260 can be used to predict the application to human patients of small molecule drug kinetics.

[0165] This embodiment is merely illustrative, and it should be understood that various modifications are possible without departing from the scope of the present invention.

Claims

1. A fluid device, The entrance and Exit and The system comprises a first group of sub-rooms, including two or more first sub-rooms arranged parallel to each other between the entrance and the exit, Each of the first sub-compartments is configured to receive each corresponding cell culture insert, The fluid flow between the inlet and the outlet is distributed evenly and / or is substantially identical between the first sub-chambers. Fluid equipment.

2. A fluid apparatus according to claim 1, wherein the inlet is in fluid communication with the first group of sub-chambers, and / or the outlet is in fluid communication with the first group of sub-chambers.

3. A fluid apparatus according to claim 1 or 2, configured to enable fluid flow between the inlet and outlet, each of the first sub-chambers separately.

4. A fluid apparatus according to any one of claims 1 to 3, wherein the flow rate of the fluid between the inlet and the outlet is substantially the same and / or distributed equally through each of the first sub-chambers.

5. A fluid apparatus according to any one of claims 1 to 4, comprising a second group of sub-chambers including one or more second sub-chambers arranged parallel to the inlet and the outlet.

6. A fluid apparatus according to claim 5, wherein the flow rate of the fluid between the inlet and the outlet is substantially the same through each of the second sub-chambers.

7. A fluid apparatus according to claim 5 or 6, wherein the second group of sub-chambers is provided between the inlet and the first group of sub-chambers, or between the first group of sub-chambers and the outlet.

8. A fluid apparatus according to any one of claims 5 to 7, wherein each sub-chamber of the second group of sub-chambers is in direct fluid communication with one or more first sub-chambers of the first group of sub-chambers via each corresponding first fluid channel.

9. A fluid apparatus according to any one of claims 5 to 8, wherein two or more first subchambers of the first group of subchambers are in direct fluid communication with the outlet via their respective corresponding outlet fluid channels, and one or more second subchambers of the second group of subchambers are in direct fluid communication with the inlet via their respective corresponding inlet fluid channels.

10. A fluid apparatus according to any one of claims 5 to 8, wherein two or more first subchambers of the first group of subchambers are in direct fluid communication with the inlet via their respective corresponding inlet fluid channels, and one or more second subchambers of the second group of subchambers are in direct fluid communication with the outlet via their respective corresponding outlet fluid channels.

11. A fluid apparatus according to any one of claims 5 to 10, wherein each of the inlet fluid channels is of the same size and / or is provided symmetrically with respect to one another.

12. A fluid device according to any one of claims 5 to 11, wherein each of the outlet fluid channels is of the same size and / or is provided symmetrically with respect to one another.

13. A fluid device according to any one of claims 5 to 12, wherein each of the first fluid channels is of the same size and / or is arranged symmetrically with respect to one another.

14. A fluid apparatus according to any one of claims 1 to 13, wherein the size and / or configuration of the sub-chambers within each group are substantially identical.

15. A fluid apparatus according to any one of claims 1 to 14, wherein the channel and the sub-chamber define a flow path between the inlet and the outlet.

16. A fluid apparatus according to any one of claims 1 to 15, wherein the sub-chamber is configured to receive a cell culture insert selected from a 24-well insert, a 12-well insert, or a 6-well insert.

17. The entrance and Exit and A first group of sub-rooms including two or more first sub-rooms arranged parallel to each other between the entrance and the exit, A second group of sub-rooms includes one or more second sub-rooms arranged parallel to the entrance and the exit, A fluid apparatus comprising one or more further groups of sub-compartments optionally arranged parallel to the sub-compartments of the first group and the sub-compartments of the second group, The inlet is in fluid communication with the first group of sub-chambers or the second group of sub-chambers via each of the (multiple) corresponding inlet channels. The outlet is in fluid communication with the second group of sub-chambers or the first group of sub-chambers via each of the (multiple) corresponding outlet channels. Each sub-compartment of a group is in fluid communication with one or more sub-compartments of an adjacent group via its corresponding fluid connection channel. Each sub-compartment is configured to receive its corresponding cell culture insert. Each of the inlet fluid channels is substantially the same in size and / or arranged symmetrically to one another. Each of the outlet fluid channels is substantially the same in size and / or arranged symmetrically to one another. A fluid apparatus in which each of the fluid connection channels between adjacent sub-compartments of a group of sub-compartments is substantially identical in size and / or is arranged symmetrically with respect to one another.

18. A fluid apparatus according to any one of claims 1 to 17, A pump that pressurizes and delivers fluid through the fluid device, A fluid system equipped with the following features.

19. A system according to claim 18, further comprising an inlet conduit configured to provide fluid communication between the pump and the inlet, and an outlet conduit configured to provide fluid communication between the pump and the outlet.

20. A system according to claim 18 or 19, further comprising a fluid supply interface configured to enable the supply of fluid to a flow path defined by the system.

21. A system according to any one of claims 18 to 20, further comprising a cell culture insert configured to be used in each corresponding sub-chamber of the fluid apparatus.

22. A method for testing the effect of a substance on one or more types of cells, A step of providing the system according to any one of claims 18 to 21, The steps include providing cell culture inserts into one or more sub-compartments of the fluid apparatus, A step of supplying fluid to the flow path defined by the system and / or the fluid device of the system, A method comprising the step of circulating the fluid so that it passes through the fluid apparatus and / or system.

23. A method for manufacturing a fluid device according to any one of claims 1 to 17, comprising the step of manufacturing the fluid device by additive manufacturing, and optionally by 3D printing.

24. A computer program having a computer-executable instruction that, when executed by a processing device, causes the processing device to control an additive manufacturing device in order to manufacture a fluid device according to any one of claims 1 to 17.

25. A method for manufacturing equipment by additive manufacturing, A step of obtaining an electronic file representing the structure of a fluid apparatus according to any one of claims 1 to 17, A method comprising the step of manufacturing a fluid device according to any one of claims 1 to 17 by controlling an additive manufacturing apparatus, through one or more additive manufacturing steps, in accordance with the structure identified by the electronic file.

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