Systems, methods and devices for culturing multicellular structure

Abstract

To provide systems, methods and devices for culturing a multicellular structure such as an organoid.SOLUTION: An exemplary system comprises a vessel, an electric / magnetic module and a control circuit. The vessel may include a culture chamber to contain a multicellular structure. The electric / magnetic module may be configured to be located in the vessel, at a position in or adjacent the culture chamber. The control circuit may be configured to wirelessly power and / or operate the electric / magnetic module.SELECTED DRAWING: None

Description

Technical Field

[0001] This application claims the benefit of priority of U.S. Provisional Application No. 62 / 976,151, filed on Feb. 13, 2020, the content of which is incorporated herein by reference in its entirety.

Background Art

[0002] Organoids ( "mini-organs") are three-dimensional aggregates of different types of cells that are generated in vitro and have a certain degree of similarity to organs, such as presenting a realistic histology of organ-specific tissues. The cell aggregates can be generated by seeding a small number of stem cells into a matrix. The stem cells then proliferate, differentiate, and self-organize within the matrix while using the matrix as a scaffold. In this approach, organoids similar to tissues from, among other things, the brain, heart, intestine, kidney, liver, and stomach have been generated to date. These promising results suggest that organoid culture has the potential to provide new insights into organ development and function and to reproduce disease models that enable drug screening in vitro. Organoids can revolutionize the way drugs are discovered and medications are personalized.

[0003] Despite the growing importance of organoids, they remain challenging to culture efficiently. New systems, methods, and devices for culturing organoids and other multicellular structures are needed.

Summary of the Invention

Means for Solving the Problems

[0004] The present disclosure provides a system, method, and device for culturing multicellular structures such as organoids. An exemplary system includes a container, an electric / magnetic module, and a control circuit. The container may include a culture chamber for containing the multicellular structure. The electric / magnetic module may be configured to be located within the container, within or adjacent to the culture chamber. The control circuit may be configured to wirelessly power and / or operate the electric / magnetic module. The present invention provides, for example, the following items. (Item 1) A system for culturing a multicellular structure, a container including a culture chamber configured to contain the multicellular structure, an electric / magnetic module configured to be located within the container, within or adjacent to the culture chamber, and a control circuit configured to wirelessly power and / or operate the electric / magnetic module comprising the system. (Item 2) The system according to item 1, wherein the control circuit is configured to wirelessly transmit power to the electric / magnetic module via inductive coupling or capacitive coupling. (Item 3) The system according to any one of items 1-2, wherein the electric / magnetic module is contained within or configured to be contained within the culture chamber, includes a magnet, and the control circuit is configured to create a magnetic field that drives the movement of the magnet within the culture chamber. (Item 4) The system according to item 1, wherein the electric / magnetic module includes a chemical sensor, an electrical sensor, an optical sensor, and / or a temperature sensor. (Item 5) The system according to any one of items 1-4, wherein the electric / magnetic module includes a pump configured to drive fluid flow into and / or out of the culture chamber. (Item 6) The electrical / magnetic module is located or configured to be located within a slot adjacent to the culture chamber, and optionally, the container defines an opening providing fluid communication between the slot and the culture chamber, the system according to any one of items 1-5. (Item 7) The container includes two or more reservoirs in fluid communication with the culture chamber, the slot is located vertically above the culture chamber, optionally between at least a pair of the two or more reservoirs, and optionally, each of the two or more reservoirs communicates separately with the culture chamber, the system according to item 6. (Item 8) The system further comprises a container assembly interconnected to form a row of containers including the container, each container in the row of containers includes an individual culture chamber for containing a multicellular structure, and optionally, the containers in the row of containers are substantially identical to each other, the system according to any one of items 1-7. (Item 9) The system further comprises a frame for holding a plurality of container assemblies including the container assembly, the frame optionally has a length and / or width corresponding to the length and / or width of a standard microplate footprint, the system according to any one of items 6-8. (Item 10) A method of culturing a multicellular structure, comprising containing a multicellular structure within a culture chamber of a container, wherein an electrical / magnetic module is located within the container, within or adjacent to the culture chamber, using a control circuit to wirelessly power / operate the electrical / magnetic module and including. (Item 11) Powering / operating includes wirelessly transmitting electrical energy to the electrical / magnetic module, the method according to item 10. (Item 12) The method according to item 10 or 11, wherein powering / operating is at least partially implemented via inductive or capacitive coupling between the control circuit and the electrical / magnetic module. (Item 13) The method according to any one of items 10 - 12, wherein the electrical / magnetic module includes a magnet, and powering / operating includes driving the movement of the magnet within the culture chamber using a magnetic field created by the control circuit. (Item 14) The method according to any one of items 10 - 13, wherein powering / operating includes sensing the properties of the multicellular structure and / or the culture medium within the culture chamber using a sensor of the electrical / magnetic module. (Item 15) The method according to any one of items 10 - 14, wherein powering / operating includes driving fluid flow into and / or out of the culture chamber. (Item 16) The method according to any one of items 10 - 15, further including collecting data related to the multicellular structure while the multicellular structure remains within the culture chamber. (Item 17) The method according to item 16, wherein collecting data is implemented using a sensor of the module. (Item 18) A device for culturing a multicellular structure such as an organoid, the device comprising: A shell having an open top; An inserter including two or more reservoirs, the inserter being configured to be received within the shell via the open top such that the shell and the inserter cooperatively form a culture chamber for the multicellular structure, the culture chamber being located below the two or more reservoirs and in fluid communication with each of the two or more reservoirs via individual channels defined by the inserter; A device comprising the above. (Item 19) The device according to item 18, further comprising a scaffold configured to assist in the formation of organoids within the culture chamber. (Item 20) The device according to item 19, wherein the scaffold is provided by a module configured to be located within or in a slot defined by the inserter. (Item 21) The device according to any one of items 18 - 20, wherein the shell includes a section having an upper region defining a receiving space and a lower region forming a receptacle, the inserter is configured to be received within the receiving space, and the receptacle is configured to cooperatively form the culture chamber with the inserter. (Item 22) The device according to any one of items 18 - 21, wherein the inserter forms a slot, defines an opening at the bottom end of the slot, and further includes a module configured to be located vertically above or in the slot with respect to the opening. Optionally, the module is an electrical / magnetic module. (Item 23) A system for culturing a multicellular structure, the system comprising: A container including a culture chamber for containing the multicellular structure, and two or more reservoirs and slots each in fluid communication with the culture chamber; Two or more modules having different functions from each other and configured to be interchangeably installed within the slot. A system comprising the above. (Item 24) The system according to item 23, wherein the container defines an opening at the bottom end of the slot in communication with the culture chamber. (Item 25) The system according to any one of items 23 - 24, wherein the two or more modules include a first module having a permeable membrane. Brief Description of the Drawings

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[0046] Detailed Description The culturing of large organoids (e.g., up to about 4 mm) is labor-intensive, complex work, and can easily require several months until the organoids can be harvested or used (e.g., for screening). During culturing, the organoids pass through different stages of the culturing protocol as they grow, differentiate, and develop. Currently, most of these stages require manual interaction for feeding, monitoring, processing, etc. The culture vessels containing the organoids may not be suitable for monitoring the state of the developing organoids, so the replacement of experimental instruments is often necessary. Culture protocols that rely on manual interaction are expensive and prone to process errors. Also, to culture large organoids that develop specific interacting cell types, it may be essential not only to treat the cells with appropriate compounds but also to expose the organoids to a suitable physical environment specific to the type of organoid being cultured. For example, cardiomyocytes as well as neurons need to be stimulated by electrical pulses, while muscle organoids and bone organoids rely on the application of alternating mechanical strain. Organoids can be cultured using complex apparatuses that rely on tubing and wiring. However, these apparatuses are bulky, not standardized, and not easily scalable for culturing multiple organoids simultaneously.

[0047] The present disclosure addresses, in various aspects, the lack of automation, non-standardized apparatuses, the presence of tubing and wiring, and non-scalability in organoid culturing as currently practiced. More specifically, the present disclosure provides containers, modules, and control circuits that enable automation, improve standardization, avoid tubing and wiring, and enable scalability.

[0048] The present disclosure provides a system and method for culturing multicellular structures such as organoids. The system may comprise a container including a culture chamber for containing the multicellular structure. An electrical / magnetic module may be configured to be located within the container, in or adjacent to the culture chamber. A control circuit may be configured to power and / or operate the electrical / magnetic module by wireless transmission. In the method, the multicellular structure may be contained within the culture chamber of the container. The electrical / magnetic module may be located within the container, in or adjacent to the culture chamber. The electrical / magnetic module may be powered / operated using the control circuit via wireless transmission of power, force, and / or signals (e.g., data). The electrical / magnetic module may include, for example, a magnet that can be moved or is moved within the container by a magnetic field created by the control circuit such that the module pumps fluid or mechanically stimulates the multicellular structure contained within the culture chamber. In other embodiments, the electrical / magnetic module may include electrodes for stimulating the multicellular structure, sensors for sensing the properties of the multicellular structure and / or the culture medium within the container, light sources for illuminating at least a portion of the multicellular structure, and / or the like.

[0049] The ability to wirelessly power / operate the electrical / magnetic module in the present system and method is significant as it eliminates the need for wires or other electrical conductors that extend from the control circuit to the container. As a result, the container becomes more portable with fewer restrictions regarding the location where it is positioned relative to the control circuit and can be more easily separated from the control circuit (e.g., to perform an imaging procedure). Also, this wireless approach can improve the fidelity and stability of experiments involving multicellular structures as it allows the electrical / magnetic module to be positioned very close to or in contact with the multicellular structure being analyzed. The present system and method can also facilitate the automation of complex culturing of multicellular structures by providing an essential physical environment and enable multi-channel in-situ monitoring of multicellular structures. The present system and method may be capable of simultaneously stimulating, maintaining, and monitoring a large number of organoids in an automated manner.

[0050] The container of the present disclosure may be customized by introducing one or more active / passive modules into one or more compartments of the container, if necessary. The selection of the functional modules and the compartments in which the modules are received enables the container to be functionally adapted during manufacture and / or by the user. For example, the selection of the modules and compartments enables, inter alia, the culturing of a specific type of organoid, the implementation of a particular culture protocol or protocol stage, the creation of desired test conditions, and / or the on-board (within the container) sensing, measurement, and / or monitoring of desired parameters. Thus, while the basic structure of the container can be standardized, the functionality of the container can be modified by the introduction of different modules to suit the specific needs of various users, if necessary. Further, the introduction of modules into the container allows the modules to come very close to or in contact with the multicellular structure, which creates a more direct interaction between the module and the multicellular structure. Additionally, the introduction of modules into the container may not result in any increase in the size of the footprint. Thus, the container modified with modules can remain compact, which allows more copies of the container to fit within the footprint of a standard microplate, which, in turn, allows more multicellular structures to be cultured simultaneously in an incubator.

[0051] Devices and methods for culturing multicellular structures such as organoids are provided. The device may comprise a shell having an open top. The device may also comprise an inserter including two or more reservoirs. The inserter may be configured to be received within the shell through the open top such that the shell and the inserter cooperatively form a culture chamber for the multicellular structure. The culture chamber may be located below two or more of the reservoirs and may be in fluid communication with each of the two or more reservoirs through individual channels defined by the inserter. In the method, an inserter including two or more reservoirs may be placed within the shell, and the inserter and the shell may be used cooperatively to form a culture chamber. The culture chamber may be located below and in fluid communication with each reservoir of the two or more reservoirs. The multicellular structure may be cultured within the culture chamber.

[0052] The devices and methods described in the preceding paragraphs can provide various advantages for culturing multicellular structures such as organoids, including any of the following combinations. The inserter may be selected from a set of inserters having mutually different characteristics such as different reservoir, channel, and / or slot configurations. Thus, the same shell can be assembled with different types of inserters to customize the resulting container structure for the specific needs of the user. Also, the inserter may position reservoirs (and optional slots) vertically above the culture chamber such that gravity can drive fluid flow into and / or out of the culture chamber. Further, the shell may define rows of compartments each capable of receiving an inserter to form individual culture chambers. Thus, the shell and two or more inserters can be assembled to form a container assembly having rows of culture chambers. Further, two or more of the container assemblies may be held by a frame having a footprint corresponding to that of a standard microplate, enabling a compact array of containers to be formed. Each container assembly may be individually removable from the frame for processing separately from the other container assemblies of the array.

[0053] The container may provide a plurality of reservoirs that are in fluid communication with the chamber via channels and that may be formed within a shared wall between the reservoir and the chamber. This configuration may be described as a standard feeding interface. In some embodiments, 3D printing provides connections to any suitable printed structure of the standard feeding interface inside the container, enabling the growth of different types of organoids.

[0054] The matrix can provide a temporary scaffold for the appropriate type of cells as they develop into organoids. The cells may self-organize and generate their own extracellular matrix, which may replace some or all of the scaffold. The same may apply to internal feeding, and the container may provide a general interface, which may optionally be modified by 3D printing, and the cells may self-organize to best utilize this modified interface.

[0055] In some embodiments, the scaffold (with or without cells) may be placed within a receptacle of the container body, and the culture chamber may optionally be formed from the receptacle using a sealing member while the container is inverted. Once these processes are completed, the container may be swiveled with the right side up (relative to its organoid culture orientation), and at least one reservoir above the culture chamber may be filled with a feeding solution. If no cells are yet present inside the scaffold, suitable cells may be placed in the feeding solution and introduced into the scaffold along with the feeding solution from the reservoir above the culture chamber.

[0056] The organoids formed may require an initial culture time before a specific feeding protocol can be initiated. The feeding protocol may involve filling the reservoirs with a suitable medium and removing the medium from the reservoirs according to a predetermined schedule and / or based on the development stage or conditions of the organoids. The feeding protocol may depend on the shape of the scaffold and the type of organoid to be formed.

[0057] The container may enable light-sheet 3D imaging. The culture chamber of the container may have two, three, or more optical windows, and light may propagate into and / or out of the culture chamber through each optical window. For example, the container may have a bottom window and one or more side windows, each of which may be planar. In some embodiments, the container may have a pair of side optical windows arranged opposite each other.

[0058] The present disclosure enables the generation of large functional organoids. Large organoids may be greater than, among other things, about 0.1, 0.2, 0.5, 1, or 2 millimeters in average or maximum diameter. Working with large organoids remains difficult, and researchers face two main limitations. First, each type of organoid may require different culture conditions, such as a specific hydrogel as a scaffold or even mechanical stimuli such as shear forces from media flow. Second, microscopy of large organoids can be very difficult. State-of-the-art methods still involve sectioning, staining, and imaging fixed samples using confocal scanning microscopy or even a slide reader of the organoid material.

[0059] The present disclosure provides systems, methods, and devices for improved organoid culture. By using a combination of 3D printing (scaffold and / or cells) and medium exchange by gravity flow, a user may generate a unique 3D environment optimized for each type of organoid. A wide range of different organoid types may be grown. Feeding and waste removal may be addressed by fluid communication between a culture chamber of a container and a reservoir. Integrating optical windows into each container, at least one for the incidence of excitation light and another for the emission of emitted light, enables monitoring of live cells of the organoid by light sheet microscopy. Alternatively, or in addition, the organoid may be imaged by classical wide-field microscopy through one or more than one of the optical windows. Thus, the containers disclosed herein may enable the performance of live cell microscopy of developing and / or developed organoids. High content and / or high throughput microscopy may be performed on the organoids.

[0060] Further aspects of the present disclosure are described in the following sections, namely, (I) Definitions, (II) Overview of the culture system and method, (III) Electrical / magnetic modules, (IV) Passive modules, (V) Container assembly, and (VI) Selected aspects. I. Definitions

[0061] Technical terms used in the present disclosure have meanings generally recognized by those skilled in the art. However, the following terms may be further defined as follows.

[0062] Cell - The basic structural, functional, and biological unit of a living organism. A cell can be eukaryotic or prokaryotic. Exemplary cells include stem cells, differentiated cells, immortalized cells (e.g., cell lines), primary cells, cells from tissue samples, transfected cells, cells from clinical samples (e.g., blood samples, fluid aspirates, tissue explants, etc.), cells forming an entire organism, and / or equivalents.

[0063] Any suitable cells may be introduced into the culture chamber (or into a receptacle that would form part of the culture chamber). The cells introduced may include stem cells (e.g., pluripotent stem cells), supporting cells, and / or the like. The cells may be deposited into the culture chamber or receptacle, and / or onto or into a scaffold located within or to be located within the culture chamber or receptacle, by any suitable technique including pipetting, bioink droplet printing, microcontact printing, photolithography, dip-pen nanolithography, and / or the like.

[0064] Cell culture - promoting the survival, health, growth, proliferation, differentiation, and / or self-organization of living cells such as cells of a multicellular structure in an artificial environment.

[0065] Culture chamber - a compartment containing a multicellular structure and having walls on substantially all sides of a mostly or completely enclosed space. At least one of the walls may define one or more openings to allow for communication with and / or passage into and / or out of the compartment.

[0066] Culture medium - an aqueous composition for cell culture. The composition may be liquid or semi-solid. The composition may include, inter alia, a carbon source (e.g., glucose), inorganic salts, vitamins, and growth regulators. The term "medium" as used herein means at least one medium and may refer to, for example, separate volumes of medium, a first medium and a second medium of different compositions, a medium of substantially the same composition in contact with different / separate cell cultures, or a combination / mixture of previously separated volumes of the same medium.

[0067] Culture vessel - A device for culturing multicellular structures. A culture vessel (synonymously called a vessel) may include a culture chamber and one or more reservoirs in fluid communication with the culture chamber. A vessel assembly or vessel array is a collection of culture vessels for culturing a one-, two-, or three-dimensional array of multicellular structures. The culture vessels disclosed herein may be single-use devices (consumables) or may be reused.

[0068] Exemplify - To be illustrative of or to serve as an example. Similarly, the term "exemplify" means to illustrate by giving an example. Neither term implies desirability or superiority or inferiority.

[0069] Inside - When describing the location / position of an object relative to a given structure, "inside" or "interior" means that the object is at least predominantly (more than 50% of the volume of the object) or completely inside the given structure. In the same context, "outside" means that the object is at least predominantly (more than 50% of the volume of the object) or completely outside the given structure.

[0070] Light - Optical radiation including ultraviolet radiation, visible radiation (i.e., visible light), and / or infrared radiation.

[0071] A module is a structurally and functionally discrete unit configured to be contained within a culture vessel. The module may be insertable into the culture vessel while the culture vessel remains intact or only when the culture vessel is disassembled, and / or may be removable therefrom, or may be configured to be non-removable from the culture vessel. The module may be an active module, also referred to as an electro / magnetic module, which utilizes electricity and / or magnetism for operation and is optionally powered / operated by wireless transmission between a control circuit and the module. Alternatively, the module may be a passive module, which does not utilize either electricity or magnetism to fulfill its intended purpose. The electro / magnetic module includes a magnetic module with a permanent magnet but no electrical / electronic devices, an electrical module with electrical / electronic devices but no permanent magnets, and a module with both a permanent magnet and electrical / electronic devices. The magnetic module may require an externally generated, optionally time-varying magnetic field for operation (such as to drive the movement of the magnetic module and / or its magnet).

[0072] The module may have any suitable shape and size. The module, particularly its housing or body, may be, for example, cubic (e.g., a cube), cylindrical, conical, or the like. The shape of the module in cross-section may correspond to that of a slot or other compartment in the container such that the module fits into the slot or other compartment. For providing flexibility and interchangeability, it may be advantageous for all modules configured to be installed in a given compartment (e.g., its slot) of the container to have the same standard shape and size, or for all modules to have the same standard size and shape regardless of the destination of the compartment. 4×4×4mm 3 is an exemplary size for both active and passive modules, which corresponds to the size of large organoids that can be generated within the culture chamber of the container.

[0073] Multicellular structure - A three-dimensional array of biologically connected cells. The multicellular structure may be an organized multicellular structure consisting of different cell types arranged non-randomly relative to each other. Exemplary multicellular structures include organoids, organisms (at any stage of development), tissue explants, tumors, or the like.

[0074] Near Field Communication (NFC) - Wireless communication between electronic devices using near-field radiation and inductive or capacitive coupling. Near Field Communication may be implemented, among other things, when the electronic devices are less than 50, 20, 10, or 5 centimeters from each other.

[0075] Near-field radiation - Electromagnetic radiation at a position within 50, 20, 10, or 5 centimeters from a radiation source, typically radio waves (e.g., microwaves), such as within 10, 5, or 2 wavelengths from the radiation source.

[0076] Organoid - A three-dimensional aggregate of different types of cells generated in vitro and having a certain degree of similarity to an organ, such as presenting a realistic histology of organ-specific tissue. The cell aggregate can be generated by seeding a small number of stem cells on a scaffold (i.e., matrix). The stem cells then proliferate, differentiate, and self-organize within the scaffold.

[0077] Receptacle - Optionally, a container having an open side, such as an open upper side, open bottom side, or open side. The receptacle may be converted into a culture chamber by at least partially covering or closing the side of the receptacle.

[0078] Scaffold - an extracellular support framework for culturing multicellular structures. The scaffold is typically a matrix in which the cells of the multicellular structure are embedded or will be embedded. The scaffold may be provided by one or more hydrogels. Each hydrogel may contain one or more thermoplastic structural components such as Matrigel, alginic acid, nanofibrillar cellulose, collagen, fibrin, and / or polyethylene glycol, which cooperatively form the matrix in a temperature - dependent manner.

[0079] In some embodiments, two or more different hydrogels / matrices may be placed within the culture chamber of a container. The hydrogels / matrices may differ with respect to any suitable parameters such as melting temperature, resistance to enzymatic degradation, solubility, cell - attracting and / or cell - repelling properties, and / or the like.

[0080] Each hydrogel / matrix may contain any suitable components. Exemplary components include one or more polysaccharides (e.g., glycosaminoglycans (GAGs) such as chondroitin sulfate, dermatan sulfate, heparin, heparan sulfate, hyaluronic acid, keratan sulfate, etc.), proteoglycans (e.g., aggrecan, agrin, brevican, collagen type XVIII, perlecan, neurocan, versican, or GAGs attached (such as via its serine) to a core protein to form a small leucine - rich proteoglycan or the like), fibrous proteins (e.g., collagen, elastin, fibronectin, laminin, etc.), and / or the like. Protease recognition sites (e.g., for matrix metalloproteinases (MMPs)) may be incorporated into the hydrogel / matrix to enable degradation / remodeling by cells. The frequency of such sites, along with the sequence of each site, may be selected to enable a suitable amount of degradation / remodeling.

[0081] One or more growth factors may be included in the matrix when formed, or introduced into the culture medium after matrix formation. Exemplary growth factors that may be suitable include angiopoietin, bone morphogenetic protein (BMP), ciliary neurotrophic factor, colony stimulating factor, ephrin, epidermal growth factor, erythropoietin, fibroblast growth factor, glial-derived neurotrophic factor, hepatocyte growth factor, insulin, insulin-like growth factor, interleukin, leukemia inhibitory factor, keratinocyte growth factor, neuregulin, neurotrophin, platelet-derived growth factor, transforming growth factor, tumor necrosis factor (alpha), vascular endothelial growth factor, and / or equivalents. II. Overview of Culture Systems and Methods

[0082] This section provides an overview of the culture systems and methods of the present disclosure (see FIGS. 1-7).

[0083] FIG. 1 shows an exemplary culture system 100 for forming, growing, differentiating, organizing, stimulating, sensing, analyzing, and / or imaging a multicellular structure such as an organoid. The culture system 100 comprises a container array 101 including a set of culture vessels 102, only three of which are clearly identified in FIG. 1. Each culture vessel 102 is configured to contain an individual multicellular structure. The container array 102 can be, or can include, a linear array, a two-dimensional array (e.g., a rectangular array, a hexagonal array, etc. as shown), and / or a three-dimensional array. Each of the container array 101 and / or two or more discrete container assemblies thereof has an occupancy area corresponding to the length and / or width of a standard microplate to facilitate mechanical and fluid handling using a robotic system designed to manipulate standard microplates. The number of culture vessels 102 in the container array 101 is at least three in a first dimension and one or more (e.g., at least two, three, or more) in a second orthogonal dimension.

[0084] Each culture vessel 102 of the container array 101 may include two or more distinct compartments, which may or may not be in fluid communication with each other and may or may not share one or more walls with each other (see FIG. 1A). The culture vessel 102 has a culture vessel 103 for containing a multicellular structure 104 such as an organoid. At least one reservoir 105 of the vessel 102 is configured to hold a culture medium. Each reservoir 105 is arranged in fluid communication with the culture chamber 103 via at least one individual connection channel 106. The vessel 102 may also have at least one slot 107 that can communicate with the culture chamber 103 via an opening 108.

[0085] Each compartment of the culture vessel 102 may have any suitable size and shape. The culture chamber 103 may have a volume of at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.7, or 1 mL. The culture chamber may be sized to contain a multicellular structure such as an organoid of any suitable size, especially having a diameter of at least 0.2, 0.5, 1, or 2 mm. In an exemplary embodiment, each reservoir 105 of the culture vessel 102 has a volume that is at least 2, 5, or 10 times the volume of the culture chamber, and / or a volume greater than the culture chamber 103, such as at least 0.5, 1, 2, 4, or 6 mL. The slot 107 may have a volume greater than, less than, or equal to the culture chamber 103 or one or more reservoirs 105, and / or may have the same height as each reservoir 105. Each compartment of the vessel may be rectangular, circular, elliptical, or the like in a horizontal cross-section. A rectangular cross-section for each compartment of the vessel may be advantageous as this shape makes very efficient use of the available space and allows the culture vessels 102 to be arranged very close to each other in the container array 101.

[0086] The culture vessel 102 may be formed from any suitable material by any suitable procedure. In an exemplary embodiment, the culture vessel may consist of at least one polymer, which may include a transparent polymer. The culture vessel 102 may be integrally formed as a single piece or may be formed by at least a pair of discrete components such as a shell and an inserter or a body and a seal member that are fitted together to form the culture chamber 103 and / or attached to each other. Thus, the culture chamber 103, the reservoir 105, and / or the slot 107 may have fixed positions relative to each other and / or may be non-removably / firmly attached to each other within the culture vessel 102.

[0087] The reservoir 105 of the container 102 may hold any suitable substance to be supplied to the culture chamber 103. Exemplary substances include, inter alia, nutrients, effectors, and reagents. Suitable nutrients include any substance for promoting the health and growth of cells inside the culture chamber 103 and thus the growth and development of multicellular structures such as organoids. Exemplary nutrients may include saccharides (such as glucose), amino acids, proteins, nucleotides, vitamins, minerals, fatty acids, and the like. Effectors include any molecule (such as an inducer or inhibitor) that activates, controls, or inactivates a process or action (such as differentiation, protein synthesis, migration, etc.). Exemplary effectors include anti-cancer compounds, growth factors, differentiation factors, oligonucleotides, mRNA, or equivalents. Reagents include any compound for facilitating the analysis of multicellular structures such as organoids. Exemplary reagents include, inter alia, labels, solidifying agents, and clarifying agents. Labels may include dyes (such as visible stains and / or photoluminescence dyes). A photoluminescence dye is any substance that emits light in response to irradiation accompanied by electromagnetic radiation such as excitation light.

[0088] Each reservoir 105 may have an open top to facilitate the introduction and removal of fluid using a fluid transfer device (e.g., a pipette). A lid may be provided for placement on the culture vessel 102 to cover the open top of each reservoir 105 during culturing within the incubator. The lid may overlap vertically over the upper region of each reservoir and may optionally have a flange configured to limit lateral movement of the lid when covering the reservoir without creating an interference fit. In some embodiments, the lid may be a cap that forms a liquid-tight seal at the top of one or more than one reservoir 105.

[0089] The container 102 may contain at least one module 109 in one or more than one of the compartments of the container. FIG. 1A shows a module 109 located within the culture chamber 103, with alternative positions that are considered possible for the same module shown in phantom lines. More specifically, each module 109 may be located, among other places, within the culture chamber 103, the reservoir 105, or the slot 107. Thus, each module 109 may be located within or adjacent to the culture chamber 103. Each module 109 may be contained within the culture vessel 102 or may be located outside the culture vessel 102 and be present in a set of functionally different modules 109 configured to be selectively placed into the culture vessel 102 by a user. Each module 109 may independently be an active electrical / magnetic module (e.g., among others, an electrode module, an actuator module, a sensor module, a pump module, and / or an illumination (light-emitting) module) or a passive module (e.g., a permeable interface module, a scaffold module, or a dummy module), as further described below.

[0090] The culture system 100 may also include an incubator 110 for containing the vessel array 101 (see Fig. 1). The incubator may be temperature-controlled to a suitable culture temperature for the multicellular structure 104, such as at least 25 °C, 30 °C, or 35 °C, among others (see also Fig. 1A). The humidity and / or atmosphere inside the incubator 110 may also be controlled to promote the growth and development of the multicellular structure 104.

[0091] The culture system 100 may further include a fluid transport system 111, a detection system 112, and a conveyor 113, each of which may be located inside or outside the incubator 110. The fluid transport system 111 is configured to add liquid to and / or remove liquid from each culture vessel 102 of the vessel array 101, such as inside and / or outside each of its reservoirs 105. The fluid transport system may thus include one or more pipettes, fluid supplies, and / or waste containers. The detection system 112 is configured to collect data related to the multicellular structures contained by the vessel array 101, such as by optical detection. For example, the detection system 112 may include a light source 114 for irradiating at least a portion of the multicellular structure within each culture vessel 102 and an image sensor 115 for capturing an image of at least a portion of the multicellular structure. The conveyor 113 may be configured to move the components of the culture system 100 relative to each other. For example, the conveyor 113 may be configured to move the vessel array 101 as a unit or only a portion thereof within the culture system 100. For example, the conveyor 113 may be configured to move the vessel array 101 or its culture vessels 102 into and / or out of the incubator 110, to and / or from the fluid transport system 111, and / or to and / or from the detection system 112. Alternatively, or in addition, the conveyor 113 may be configured to remove and replace one or more lids covering the vessel array 101, if present, when the reservoirs 105 and / or slots 107 are being accessed.

[0092] The control circuit 116 of the culture system 100 powers and / or operates any suitable device of the culture system. For example, the control circuit 116 may control each of the fluid transport system 111, the detection system 112, and / or the conveyor 113 via wired or wireless communication, which can be one-way or two-way communication. Each module 109 held by the container array 101 may also be wirelessly controlled by the control circuit 116, as indicated by the dashed arrows at 117. This wireless control is advantageous because it simplifies the structure of the container array 101, allows the container array to be functionally customized as needed by introducing appropriate modules, eliminates the need for wires or electrical conductors that extend into the culture vessels and provide a path for microbial contamination.

[0093] FIG. 2 shows an exemplary container array 201 for the culture system 100 of FIG. 1. The container array comprises a plurality of container strips 218 (synonymously called container assemblies), each including a row of culture vessels 202 attached to each other, for example, via a common (shared) housing. The container strips may have any suitable number of culture vessels 202, such as at least two, three, four, or more. Each container strip 218 is received and held by a frame 219, optionally forming a row of container strips 218. Each container strip 218 may be installed as a unit into an individual receiving site of the frame 219, for example, during manufacture or by the user. In the depicted embodiment, the frame 219 has a row of eight receiving sites for receiving a corresponding number of container strips 218, but in other embodiments, the frame may be configured to receive at least two, three, or more container strips 218 at corresponding receiving sites. Each container strip 218 may be individually removable from the frame 219 to allow for the operation, processing, and / or analysis of the container strip (and / or its contents) separately from the other container strips 218 of the container array 201.

[0094] FIG. 3 shows another exemplary container array 301 for the culture system 100 of FIG. 1. The container array comprises a plurality of separate individual culture vessels 302 that are received and held by a frame 319. Each culture vessel 302 may be installed, for example, during manufacture or by the user, into an individual opening of the frame 319. The culture vessels may or may not be removable from the frame.

[0095] FIG. 4 shows a module set 420 of functionally different modules 409a - 409c that can be contained within the culture vessel 102 of FIG. 1A. The module set 420 may consist of any suitable number of functionally different modules, such as at least two, three, four, or more. Each module of the module set 420 may have any suitable combination of characteristics as described above with respect to module 109 or elsewhere in this specification (such as in Sections I, III, IV, and VI, etc.). Two or more modules of the module set 420 may be positioned within the same compartment of the culture vessel 102 so as to be interchangeable with respect to each other, as illustrated using phantom arrows for the installation of modules 409b and 409c within slot 107, and / or at least one module of the module set 420 may be positioned so as to be interchangeable within each of two or more compartments of the culture vessel 102, as illustrated using a pair of phantom arrows for module 409a within the culture chamber 103 or reservoir 105.

[0096] FIG. 5 shows an exemplary culture vessel 502 and an exemplary control circuit 516 for the culture system 100 of FIG. 1. The culture vessel 502 is shown in a schematic side view and includes a culture chamber 503 that contains a multicellular structure 504, i.e., an organoid 521, and an associated scaffold 522 for assisting in the formation and / or growth of the organoid. The scaffold 522 is attached to the bottom wall of the culture chamber 503 of FIG. 5, although in other embodiments it may be attached to any suitable sidewall or top wall of the culture chamber. At least a pair of reservoirs 505a, 505b are positioned across the culture chamber 503 and communicate with the culture chamber via individual channels 506a, 506b. Each reservoir is "vertically above" the culture chamber, meaning that a vertical line extends through the reservoir and the culture chamber. Each reservoir 505a, 505b holds an individual culture medium 523a, 523b, which may have the same or different compositions from each other. The culture chamber 503 also holds a culture medium, which may be supplied, at least in part, by one or both of the reservoirs 505a, 505b via one or both of the channels 506a, 506b.

[0097] The culture vessel 502 optionally defines a slot 507 that extends across the culture chamber 503. The slot 507 may be described as an access slot since the slot can communicate with the culture chamber 503 via an opening 508 at its bottom end. The opening 508 may (or may not) have a diameter that exceeds that of each of the channels 506a, 506b, such as by at least 50% or 100%. The slot 507 is configured to receive a slot module 509a, which may be installed into the slot 507 of the culture vessel 502 during manufacture or by the user. In some cases, the user may select a slot module 509a for installation into the slot 507 from a set of functionally different slot modules (see, e.g., FIG. 4). The ability to exchange slot modules of different functions allows the culture vessel 502 to be adapted to different culture / test modalities for various types of organoids or other multicellular structures. In some cases, a dummy module may be installed into the slot 507 to cover the opening 508 when the slot module is not being used within the slot.

[0098] The slot 507 may have any suitable position relative to the culture chamber 503 and the reservoirs 505a, 505b. The slot may be centered between the reservoirs, as shown, or may have a lateral position relative to the reservoirs.

[0099] The slot 507 and the reservoirs 505a, 505b may be open at their upper ends. This configuration allows for the installation of the slot module 509a into the slot 507 and the dispensing of culture media 523a, 523b into one or both of the reservoirs. Thus, the vessel 502 may include a removable lid for covering the open tops of the slot 507 and / or the reservoirs 505a, 505b. Further aspects of the lid for the culture vessel and the vessel assembly are described below in Section V.

[0100] The culture chamber 503 may contain at least one chamber module 509b. The chamber module may be installed in the receptacle 524, such as through its upper side, bottom side, or side, etc. The receptacle 524 may be converted into the culture chamber 503 by at least partially closing the upper side, bottom side, or side of the receptacle. In some cases, when the chamber module 509b is large enough, the chamber module may be confined within the culture chamber 503 until the culture chamber is opened and / or disassembled by removing its wall portion, and until it is so done.

[0101] When one or both of them are electrical / magnetic modules, the control circuit 516 is configured to wirelessly control the slot module 509a and / or the chamber module 509b, as shown at 517 (see Section III). The control circuit may include one or more antennas 525 to transmit / receive power and / or data to / from each module 509a, 509b that is an electrical module, if present, using near-field radiation. One or more coils 526 of the control circuit may be utilized to generate a magnetic field to drive the movement of at least the magnetic portion of each module 509a, 509b, if present. The control circuit 516 may also include a computer including a processor 527, one or more controllers 528, a memory storage device 529, and / or a user interface 530 (e.g., a display, keyboard, mouse, printer, and / or the like).

[0102] The control circuit 516 may be configured to control the movement and / or operation of one or more electrical / magnetic modules contained within each culture vessel of an array of culture vessels. Thus, the control circuit may have at least one individual antenna 525 and / or at least individual coils 526 for each culture vessel of the array.

[0103] FIG. 6 is a flowchart 630 of exemplary steps 631a - 631j that can be implemented in any suitable order and combination to provide a method for culturing a multicellular structure. The method may be implemented using any suitable system, device, cell, and scaffold of the present disclosure.

[0104] The scaffold may be placed in the receptacle of the culture vessel or within the culture chamber in step 631a. The scaffold may be placed by forming the scaffold within the receptacle or culture chamber, such as by 3D printing, or by installing a pre - formed scaffold into the receptacle or culture chamber. In some embodiments, the scaffold may be attached to a scaffold module that can be placed within a slot of the culture vessel, either before or after the culture chamber is formed.

[0105] Cells may be introduced into the receptacle of the culture vessel or into the culture chamber in step 631b. The cells may include stem cells that are intended to generate a multicellular structure organized by differentiation, division, migration, etc. In other cases, the cells may be introduced into the receptacle or culture chamber as a pre - formed multicellular structure (e.g., an organism, tissue explant, tumor, organoid, or the like). The cells may be introduced during step 631a, or before or after step 631a.

[0106] One or more modules may be placed in or adjacent to the receptacle or culture chamber in step 631c. Any combination of modules as disclosed herein may be placed. Any suitable number of one or more modules may be placed during the manufacture of the container and / or by the user.

[0107] The culture chamber of the culture vessel may be formed in step 631d. The culture chamber may be formed at least partially by using a receptacle that may have an open side and closing the open side. For example, the open side may be covered by joining a sealing member to the receptacle at the open side (e.g., at the open bottom side of the receptacle) or by installing an inserter into a shell containing the receptacle. In either case, the chamber may be formed cooperatively using the receptacle and the sealing member or the inserter.

[0108] The module may be installed in the slot of the container in step 631e. The module may be installed before or after step 631d. In some cases, steps 631a and 631e may be performed together using a scaffold module that includes a pre-formed scaffold. In some cases, steps 631b and 631e may be performed together using a module that contains cells.

[0109] The culture medium may be added to one or more reservoirs of the culture vessel in step 631f. Once added, the culture medium may optionally be driven by gravity to flow from one of the reservoirs into the culture chamber. Gravity may also drive the flow of the culture medium out of the culture chamber and into a different reservoir of the culture vessel.

[0110] The container and its contents may be cultured in step 631g. The culturing may be performed at a suitable temperature in a suitable gaseous atmosphere, for any suitable length of time, such as for at least 1, 2, 3, 4, or 5 days, or for at least 1, 2, or 3 weeks, etc.

[0111] If present, each electrical / magnetic module within the culture vessel may be wirelessly controlled by the control circuit in step 631h. This control may include transmitting power, force, and / or data to the module to drive movement of the module or at least a part thereof and / or to control the operation of the module. Steps 631g and 631h may be performed simultaneously.

[0112] A multicellular structure may be formed, grown, stimulated, and / or sensed within the culture chamber in step 631i. Step 631i may be performed in response to step 631g and / or 631h.

[0113] The first module contained by the container may be removed and replaced by a second module in 631j. The first module may be removed from a slot of the container and replaced by the second module within the same slot. The first and second modules may be functionally different from each other. After the replacement of the first module with the second module, steps 631h and 631i may be repeated.

[0114] FIG. 7 is a flowchart 730 of exemplary steps 731ab, 731d, 731f, 731g, and 731i that may optionally add one or more steps from the flowchart 630 of FIG. 6 and be performed in any suitable order and combination to provide a method for culturing a multicellular structure. This method may be implemented using any suitable system, device, cell, and scaffold of the present disclosure. The steps of flowchart 730 corresponding to those of flowchart 630 have the same letter designations in both flowcharts.

[0115] The scaffold and / or cells may be placed within the shell of the container in step 731ab. More specifically, the scaffold and / or cells may be placed within a receptacle formed by a part of the shell.

[0116] The inserter of the container may be installed in the shell in step 731d to form a culture chamber. The culture chamber may be cooperatively formed by the receptacle and the shell. The receptacle may provide the bottom wall and the side walls of the chamber, and the inserter may provide the upper wall of the culture chamber. Step 731d may be performed before or after the scaffold and / or cells are placed in the receptacle.

[0117] Culture medium may be added in step 731f to at least one reservoir spanning (e.g., vertically above) the culture chamber. The inserter may provide at least one reservoir or each reservoir spanning the culture chamber.

[0118] The culture vessel and its contents may be cultured in step 731g. The culturing may be performed at a suitable temperature, in a suitable gaseous atmosphere, for any suitable length of time, as described above with respect to step 631g (see FIG. 6).

[0119] The methods of FIGS. 6 and 7 may be performed using an array of culture vessels. Thus, each step of the method may be performed, as appropriate, on the culture vessels of the array as a group or on individual culture vessels. III. Electrical / Magnetic Module

[0120] This section describes exemplary electrical / magnetic modules for use in the culture systems and methods of the present disclosure (see FIGS. 8-17). Each electrical / magnetic module is configured to be powered / operated wirelessly by a control circuit while the module is positioned within the culture vessel. The electrical / magnetic module utilizes electricity, magnetism, or a combination thereof that is wirelessly supplied or applied to the module by the control circuit to drive at least a portion of the movement of the module relative to the vessel and / or the operation of at least one electrical / electronic device of the module. The module may be configured to be contained within any of the culture vessels of the present disclosure and within any of its compartments such as slots, reservoirs, and / or culture chambers of the culture vessel.

[0121] FIG. 8 shows an exemplary electrode module 809 (i.e., an electrical / magnetic embodiment of module 109), and also shows a power antenna 825a and a communication antenna 825b of a control circuit 816. Antennas 825a, 825b are each configured to transmit power to electrode module 809 and operate it.

[0122] The electrode module 809 includes a housing 832 that supports an electrode interface 833. The electrode interface may be located on any suitable side of the housing 832, such as, among other things, the bottom side as shown, or the side thereof (e.g., when the electrode module 809 is contained within a culture chamber). The electrode interface 833 may include any suitable number of electrodes of any suitable shape. For example, the electrode interface may have a pair of flat electrodes 834a configured to touch the surface of a multicellular structure within a culture chamber and / or a pair of pin electrodes 834b configured to penetrate and extend into the multicellular structure. The electrode interface 833 may be used to electrically stimulate the multicellular structure and / or sense the electrical properties of the multicellular structure from the outside or inside. Exemplary uses for the electrode module 809 include, among other things, (a) cardiomyocytes or neurons for pacing or activation, (b) myoblasts to promote muscle differentiation and growth in three dimensions, or (c) electrically stimulating neural tissue (e.g., brain organoids) to promote axonal growth and morphological changes to affect network formation. Other exemplary uses for the electrode module 809 include sensing the electrical activity of any suitable cell / tissue, such as neurons, smooth muscle, cardiomyocytes, or skeletal muscle.

[0123] The housing 832 may contain any suitable electronic circuitry to enable wireless power supply to, communication with, and / or control of the electrode module 809. This circuitry may include a power receiver 835 having an antenna for receiving wirelessly transmitted power from the power antenna 825a of the control circuit 816. The circuitry may also include an internal communication antenna 836 for receiving signals from, and / or transmitting signals to, the external communication antenna 825b of the control circuit 816 to enable communication between the electrode module 809 and the control circuit 816. Any suitable communication protocol such as at least one near field communication (NFC) protocol may be utilized. The electronic circuitry may also include a power storage unit 837 for storing power from the power receiver 835, a controller 838, and a digital-analog converter (DAC) and pulse generator 839. The electronic circuitry of any module of Section III may be sealed inside the housing to prevent damage in case the module comes into contact with a liquid such as a culture medium.

[0124] Figure 9 shows an exemplary sensor module 909 (i.e., an electrical / magnetic embodiment of module 109) and also shows the communication antenna 925 of the control circuit 916. The antenna 925 is configured to transmit power to the sensor module 909 and operate it. Thus, a separate power antenna and power receiver may not be necessary (compare with FIG. 8).

[0125] The sensor module 909 comprises a housing 932 that supports a sensor interface 940. The sensor interface may be located on any suitable side of the housing 932, such as, among other things, the bottom side as shown, or its lateral side (e.g., when the sensor module 909 is contained within a culture chamber). The sensor interface 940 may be configured to sense and measure any suitable physical or chemical parameter, such as temperature, movement, electrical parameters (e.g., potential, current, impedance, etc.), electric / magnetic fields (e.g., using a Hall effect sensor), pH, chemical potential, oxygen or carbon dioxide concentration, chemical substances (e.g., using an electrochemical sensor), etc. The sensor module 909 may be suitable when remote sensing is difficult or not possible, or when it is advantageous to establish direct contact between the sensor interface 940 and the multicellular structure to be analyzed.

[0126] The housing 932 may contain any suitable electronic circuitry to enable wirelessly powering, communicating with, and / or controlling the sensor module 909. This circuitry may include an internal communication antenna 936 to receive signals from and / or transmit signals to the external communication antenna 925 of the control circuit 916 in order to power and operate the sensor module 909. Any suitable communication protocol may be utilized, such as at least one short-range wireless communication (NFC) protocol. The electronic circuitry may also include a controller 938, an analog-to-digital converter (ADC) 941, a memory 942, and an amplifier and sensor electronics 943.

[0127] FIG. 10 shows an exemplary pump module 1009 (i.e., an electrical / magnetic embodiment of module 109) and also shows the power antenna 1025 of the control circuit 1016. The power antenna 1025 is configured to transmit power to the pump module 1009. There may or may not be a separate communication antenna within the control circuit 1016 (compare with FIG. 8).

[0128] The pump module 1009 includes a housing 1032 containing a pump 1044 for driving fluid flow. The pump 1044 is in fluid communication with a pair of nozzles 1045a, 1045b located peripherally of the pump module 1009. The operation of the pump 1044 draws fluid into the nozzle 1045a (acting as an inlet) and pushes the fluid out from the nozzle 1045b, or vice versa if the pump is driven in reverse. The main movement of the fluid can generate turbulent flow, among other things, to enhance the flow of the culture medium in the culture vessel and / or to stimulate organoid development. The use of the pump module 1009 can be suitable when gravity-driven flow is not applicable or is inefficient (e.g., due to the nature of the medium).

[0129] The housing 1032 may contain any suitable electronic circuit for wirelessly receiving power from the control circuit 1016 and enabling the operation of the pump 1044. This circuit may include a power receiver 1035 for receiving power wirelessly transmitted from a power antenna 1025, such as by induction or capacitive coupling. The electronic circuit may also include a power storage unit 1037 and a pump drive device 1046.

[0130] FIG. 11 shows an exemplary culture system 1100 including a pair of pump modules 1009a, 1009b contained within a culture vessel 1102 (see also FIG. 10). The vessel 1102 includes a culture chamber 1103 containing organoids 1121. Three compartments, namely, a pair of reservoirs 1105a, 1105b holding individual culture media 1123a, 1123b and a slot 1107 for storing a dummy module 1109 (which is an example of the module 109 in FIG. 1) are located vertically above the culture chamber 1103. The reservoir 1105a communicates with the culture chamber 1103 via a pair of channels 1106a, 1106c, and the reservoir 1105b communicates with the culture chamber 1103 via a pair of channels 1106b, 1106d.

[0131] Pumps 1009a and 1009b are each driven by the power transmitted from the power antennas 1025a and 1025b of the control circuit 1016. Pump 1009a drives the culture medium 1123a from the reservoir 1105a into the culture chamber 1103 through the channel 1106c. Pump 1009b drives the culture medium from the culture chamber 1103 into the reservoir 1105b through the channel 1106d. Therefore, there is a net pump-driven flow of the medium from reservoir 1105a to reservoir 1105b. The pump can be driven in reverse to move the medium back from reservoir 1105b to reservoir 1105a, or this may be done via gravity-driven flow through channels 1106a and 1106b.

[0132] Figure 12 shows a culture system 1200 for mechanically stimulating the organoid 1221 or other multicellular structures. The culture system 1200 includes a container 1102 (see Figure 11), a control circuit 1216, and a pair of magnetic modules 1209a and 1209b contained within the culture chamber 1103 of the container 1102. The organoid 1221 may be positioned between the magnetic modules. The magnetic modules can function as magnetic actuators for applying alternating mechanical strain to the organoid, and the driving principle is the magnetic force applied by the control circuit 1216 to create linear driving.

[0133] The electrode module 809 may be located within the slot 1107, and both pin electrodes 834b extend into the organoid 1221 (see also Figure 8). However, as discussed below, the mechanical stimulation of the organoid 1221 provided by the movement of the magnetic modules 1209a and 1209b driven by the control circuit 1216 does not require the presence of the electrode module 809.

[0134] Each of the magnetic modules 1209a, 1209b includes a permanent magnet 1247 that can be encapsulated by a housing 1232. The housing may have a surface coating that promotes adhesion to the organoid 1221. The magnets 1247 of the magnetic modules 1209a, 1209b each have a north pole (N) and a south pole (S), and a magnetic axis that extends through both poles. The magnetic modules 1209a, 1209b may be arranged within the cell chamber 1103 such that the magnetic axes of the magnetic modules are coaxial with each other and anti-parallel as shown, creating a magnetic repulsion, or parallel, creating a magnetic attraction.

[0135] The control circuit 1216 includes at least one coil (equivalent of the fixed coils 1226a, 1226b) that can be electrically energized to create one or more additional magnetic fields. The additional magnetic fields effectively strengthen or reduce the attraction or repulsion between the magnetic modules 1209a, 1209b, thereby driving the movement of the magnetic modules towards or away from each other. Each coil 1226a, 1226b defines a coil axis that can be oriented parallel to the magnetic axes of the magnetic modules 1209a, 1209. FIG. 13 illustrates the energization of both coils 1226a, 1226b as indicated by the current arrows 1348 and the generation of the indicated magnetic field polarities in the coils. The magnetic repulsion between coil 1226a and magnetic module 1209a and between coil 1226b and magnetic module 1209b drives the movement of the magnetic modules towards each other as indicated by the movement arrows at 1349, which applies compression to the organoid 1221. In other cases, the energization of coils 1226a, 1226b may drive the magnetic modules 1209a, 1209b further apart, which may apply tension to the organoid if it is attached to both modules. In yet other cases, the magnetic modules may be used for magnetic sensing of organoid movement. The ability to apply repeated mechanical stress through the magnetic modules is particularly beneficial for certain types of organoids, such as bone organoids and muscle organoids, that may require this stress for proper development.

[0136] Figures 14 and 15 schematically illustrate a method by which the magnetic drive mechanisms of FIGS. 12 and 13 can be implemented in a culture system 1400 that includes a row of culture vessels 1402a - 1402c. (The positions of the culture vessels are roughly indicated using dashed arrows, but the vessels themselves are omitted for simplicity of illustration.) Each of the culture vessels 1402a - 1402c contains an individual organoid 1421a - 1421c sandwiched between a pair of magnetic modules 1209a, 1209b, as described above with respect to the single culture vessel 1102. The control circuit 1416 provides a series of electrically energizable coils 1426a - 1426d that are arranged along the same lines as each of the vessels 1402a - 1402c, the organoids 1421a - 1421c, and the pairs of magnetic modules 1209a, 1209b. The coil axes defined by the coils 1426a - 1426d each extend along the row of vessels 1402a - 1402c.

[0137] The positions of the magnetic modules 1209a, 1209b are shown in FIG. 14 without the electrical energization of the coils 1426a - 1426d and in FIG. 15 with it. The electrical energization is indicated using the arrows at 1448 in FIG. 15. As described above with respect to the single culture vessel of FIGS. 12 and 13, appropriate electrical energization of the coils 1426a - 1426d drives each pair of magnetic modules 1209a, 1209b linearly towards each other along the lines of the vessels 1402a - 1402c, which enables the organoids 1421a - 1421c to be mechanically stimulated synchronously.

[0138] FIG. 16 is a schematic top view of a culture system 1600 having different configurations of electrically energizable coils 1626 for driving the movement of the magnetic modules 1209a, 1209b of FIGS. 14 and 15. (Only a subset of the coils 1626 are identified using numerical identifiers.) The culture system 1600 includes a row of containers 1602a - 1602c. (The positions of the containers are roughly indicated using dashed arrows, but the containers themselves are omitted for simplicity of illustration.) Each container 1602a - 1602c contains an individual organoid 1602a - 1602c sandwiched between a pair of the magnetic modules 1209a, 1209b as described above with respect to the culture system 1400. However, the magnetic axes of each pair of the magnetic modules 1209a, 1209b are oriented orthogonally to the row of containers 1602a - 1602c, such as horizontally, as shown. The control circuit 1616 provides at least one row or a pair of rows of the coils 1626. Each coil 1626 may define a coil axis that is parallel to the magnetic axis of the magnetic module. Appropriate electrical energization of the coils 1626 drives the magnetic modules 1209a, 1209b linearly in a direction parallel to the row of containers 1602a - 1602c and mechanically stimulates the organoids 1621a - 1621c.

[0139] FIG. 17 shows an exemplary optical module 1709 (i.e., an electro - magnetic embodiment of module 109) configured to be wirelessly powered and controlled by a control circuit 1716 via its power antenna 1725. The optical module 1709 includes a housing 1732 containing a power receiver 1735 for receiving power from the power antenna 1725, a power storage unit 1737, and a light source 1750. The light source generates optical radiation, which may be used for irradiation for optical stimulation of a multicellular structure within a container (e.g., via optogenetics) and / or for optical detection of the multicellular structure.

[0140] Exemplary light sources include light emitting diodes, lasers, or the like. The light source may also include any suitable optical system for directing or focusing the light. For example, the light source may have a waveguide such as an optical fiber to direct light from the light module 1709 onto the surface of the multicellular structure and / or to extend into the multicellular structure and irradiate the multicellular structure from the inside.

[0141] The light module 1709 may be used to facilitate imaging of the multicellular structure contained within the culture chamber of the culture vessel. For example, the light module may provide brightfield illumination or darkfield illumination of the multicellular structure when appropriately positioned within the culture vessel, such as within its reservoir, slot, or within the culture chamber. IV. Passive Modules

[0142] This section describes exemplary passive modules for use in the culture systems and methods of the present disclosure (see FIGS. 18 - 20). The passive modules are configured to function without a source of power and without interaction with a control circuit. Each passive module may be configured to be placed within a particular compartment of the container or, alternatively, within each of two or more compartments of the container.

[0143] FIG. 18 is a somewhat schematic view of an exemplary scaffold module 1809, which is an embodiment of the module 109 of FIG. 1A. The scaffold module includes a main body 1851 and a scaffold 1822 attached to the main body 1851. The scaffold 1822 is configured to assist in the formation and growth of multicellular structures such as organoids. The scaffold may be formed on the main body 1851 by, for example, 3D printing, or may be first formed and then attached to the main body 1851. In either case, the scaffold 1822 may be mounted on any suitable side of the main body 1851, such as, inter alia, the bottom side, the top side, or the side side. The scaffold module 1809 may be located within a slot of a culture vessel, such as during the manufacture of the culture vessel or when installed by the user into the slot. In some cases, the scaffold module may be an inserter that forms one or more reservoirs of the culture vessel, as further described below in Section V. The scaffold module may be used to mount and support the scaffold and to keep the scaffold in place, enabling improved handling and addition and removal of the scaffold.

[0144] FIG. 19 shows an exemplary permeable interface module 1909, which is an embodiment of the module 109 of FIG. 1A. The permeable interface module is sectioned in FIG. 19 to expose its internal structure. The permeable interface module 1909 has a hollow main body 1951 that defines a cavity 1952 continuous with an inlet 1953 and an interface opening 1954. The inlet 1953 may be plug-connected to a flexible cap 1955. A permeable member 1956, such as a permeable membrane or gel, is attached to the hollow main body 1951 at the interface opening 1954 to create a permeable interface 1957 (e.g., a permeable wall) on any suitable side of the main body 1951, such as, inter alia, the bottom side, the top side, or the side side. The permeable interface 1957 optionally enables selective passage of fluids and / or small molecules into and out of the cavity 1952 through the permeable interface.

[0145] The permeable interface module 1909 may contain any suitable medium within the cavity 1952. The medium may be a gas, liquid, gel, or the like. The medium may have a different phase, composition, and / or chemical potential than the culture medium present in the culture chamber of the container. The permeable interface 1957 may be positioned in physical contact with the multicellular structure and / or the culture medium within the container. As an example, the permeable interface module 1909 may contain air (or other gas) within the cavity 1952, create a gas / liquid interface at the permeable member 1956, and enable the culture of lung organoids within the culture chamber of the culture vessel. In other examples, the permeable interface module 1909 may contain any suitable chemical substance, which can be discharged into the culture chamber of the culture vessel through the permeable member 1956.

[0146] Figure 20 shows a permeable interface module 1909 operatively positioned within the slot 1107 of the container 1102 (see also Figure 11), adjacent to the opening 1108, and spanning the lung organoid 2021. The cavity 1952 is filled with gas such that the permeable member 1956 forms a gas-liquid interface between the gas within the cavity 1952 and the liquid culture medium within the culture chamber 1103.

[0147] The passive module contained within the compartment of the culture vessel may be described as a dummy module. The dummy module may be a substitute. Like any of the modules of the present disclosure, the dummy module may function to reduce the fluid holding capacity of the container and / or block or seal the slot across the culture chamber. However, the dummy module may also provide a surface shape (concave or convex), surface chemistry / texture (e.g., hydrophilicity, microstructure, etc.), and / or functional surface that is advantageous with respect to the culture protocol. V. Container Assembly

[0148] This section describes exemplary container assemblies, container arrays formed using the container assemblies, and culture containers of the container assemblies for use in the culture systems and methods of the present disclosure (see FIGS. 21-40).

[0149] FIG. 21 shows an exemplary container array 2101 for culturing corresponding arrays of multicellular structures such as organoids. The container array 2101 comprises at least one container strip 2118 held by a frame 2119. Only one container strip 2118 is shown in FIG. 21, but the frame 2119 is configured to removably hold two, three, or more container strips 2118 that can be substantially identical to each other. In the depicted embodiment, the frame 2119 defines eight receiving sites 2158 for a corresponding number of container strips 2118 arranged in columns, although only a subset of the receiving sites 2158 may be occupied by the container strips 2118 at any given time. The frame 2119 may have an occupancy area corresponding to that of a standard microplate (the standard microplate occupancy area is 127.71 mm × 85.43 mm), and the length and width of the frame's occupancy area are each within 10% or 5% of that of a standard microplate. This correspondence between the frame 2119 and the standard microplate occupancy area achieves mechanical compatibility with incubators, analytical instruments, and handling systems for standard microplates.

[0150] The container strip 2118 forms a column of at least two, three, or more culture containers, such as the four culture containers 2102a-2102d in the depicted embodiment. Each culture container 2102a-2102d includes an individual culture chamber 2103 formed by the lower region of the culture container.

[0151] FIG. 22 shows a container strip 2118 that is removed from the frame 2119 to enable imaging of the multicellular structures contained within the culture chamber 2103. Removal of the container strip 2118 from the frame 2119 allows the container strip to be properly positioned with respect to the imaging system 2112 (or other detection system) without interference from the frame 2119 or other container strips 2118 held by the frame.

[0152] Each culture chamber 2103 has one or more optical windows formed by one or more walls of the culture chamber. Each optical window is configured to be transmissive to optical radiation such as visible light and may optionally have smooth inner and outer surfaces that are planar in order to minimize scattering of the optical radiation. In the illustrated embodiment, each culture chamber 2013 has a bottom optical window 2159a and a pair of lateral optical windows 2159b, 2159c that are arranged opposite each other across the culture chamber 2103 (see also FIGS. 24, 26, and 30).

[0153] FIG. 22 illustrates a method by which the imaging system 2112 can capture an image of the multicellular structures contained within the culture vessel 2102c. A thin section of the multicellular structure may be illuminated through the lateral optical window 2159b (and / or window 2159c) of the culture vessel using a light source 2114 that can generate a light sheet 2160. Light (e.g., fluorescence) from the multicellular structure may be collected using an objective lens 2161 that collects light propagated through the bottom optical window 2159a of the culture vessel 2102c. Thus, the imaging system 2112 may perform selective plane illumination microscopy (SPIM), which is also known as light sheet microscopy. In other embodiments, illumination may be performed through light collection from the bottom optical window 2159a and one of the lateral optical windows 2159b or 2159c. In yet other embodiments, illumination may be performed using a light module located within the culture vessel (see, e.g., Section III). In still other embodiments, the imaging system 2112 may utilize two-photon excitation microscopy, tomography, or the like.

[0154] The container strip 2118 provides individual pairs of reservoirs 2105a, 2105b and individual slots 2107 that span the culture chambers 2013 of each of the containers 2102a - 2102d (see FIGS. 21 and 22). A slot module 2109 may be located within the slot 2107 and may be any of the electrical / magnetic or passive modules disclosed herein (see, e.g., Sections I, III, and IV).

[0155] FIG. 23 shows an exploded view of the container strip 2118. The container strip includes a shell 2162 (synonymously referred to as a housing), a plurality of inserters 2163 configured to be received within the shell 2162, and a lid 2164 configured to cover the open upper side of the shell 2162 and / or the inserters 2163. In some embodiments, the lid 2164 may not be required. Individual pairs of magnetic modules 1209a, 1209b may be located within the shell 2162 under each inserter 2163 (see also Section III), although any other suitable module may be located within the shell under the inserter or no module may be located within the shell under the inserter.

[0156] The shell 2162 may be internally divided into a plurality of sections such as sections 2165a - 2165d. The sections 2165a - 2165d may be integrally formed with each other. The sections 2165a - 2165d may be directly attached to each other or may be separated from each other using individual spacer regions 2166 located intermediate each adjacent pair of sections.

[0157] Each section 2165a - 2165d of the shell 2162 includes a lower region that forms the receptacle 2167 and an upper region that forms a receiving space 2168 for one of the inserters 2163 (see FIGS. 23, 26, and 28 - 30). Each culture chamber 2103 is cooperatively formed using one of the shell 2162 and the inserter 2163, which may capture a module (in the described embodiment, the equivalent of the magnetic modules 1209a, 1209b (see FIG. 26)) within the culture chamber. The receptacle 2167 provides the sidewall of the culture chamber 2103, and one of the inserters 2163 provides the upper wall portion of the culture chamber. A scaffold for the organoid may be disposed within the receptacle 2167 of each inserter 2163, such as being formed or installed therein (e.g., attached to the bottom or sidewall of the receptacle) before the inserter 2163 is assembled with the shell 2162. In other cases, the scaffold may be disposed on the bottom side of the inserter 2163, such as being 3D printed thereon or attached after scaffold formation, before the inserter is assembled with the shell 2162. In still other cases, the scaffold may be provided by the module 2109 (see also Section IV).

[0158] Each inserter 2163 is configured to fit within one of the sections 2165a - 2165d. Thus, the outer dimensions of the inserter may correspond to the dimensions of the receiving space 2168. The receptacle 2167 has a horizontal dimension that is one or more smaller than the receiving space 2168, forms a shoulder 2169 at the bottom of the receiving space 2168, supports the inserter 2163, and may prevent further downward advancement into the section (see FIGS. 26, 29, and 30).

[0159] Each inserter 2163 may be locked within the shell 2162 by a snap - fit mechanism that engages when the inserter is seated within the shell (see FIGS. 27 and 31 - 33). The inserter may define a protrusion 2170 that is received within an individual opening 2171 defined by the sidewall of the shell 2162, or the inserter may define the opening and the shell may define the protrusion.

[0160] Each inserter 2163 defines various openings at its bottom. The openings include at least one corresponding channel of channels 2106a, 2106b at the bottom of each reservoir 2105a, 2105b (see FIG. 33). Each channel 2106a, 2106b extending from reservoir 2105a or 2105b provides fluid communication between one of the reservoirs and the culture chamber 2103 located vertically below the reservoir (see also FIG. 26). The inserter also defines an opening 2108 at the bottom end of slot 2107, providing fluid communication between slot 2107 and culture chamber 2103 (see FIGS. 26, 33, 34).

[0161] FIG. 35 shows a lid assembly 2172 for the container array 2101 (see also FIG. 21). The lid assembly 2172 includes a series of lids 2164 for covering a corresponding number of container strips 2118 held by a frame 2119 (see also FIG. 26). Each lid 2164 may fit over the upper portion of the container strip (e.g., the upper portion of the shell 2162), project downward, overlap the container strip vertically at its upper edge, and may have a peripheral flange configured to horizontally surround it. Maintaining a sterile state can be a major problem during the entire process of culturing and testing, which can take up to several months. Therefore, a lid assembly 2172 for blocking the entry of contaminating microorganisms through the open upper portions of the reservoirs 2105a, 2015b and slots 2107 of each container 2102a - 2102d of each container strip 2118 would be advantageous. The lid assembly 2172 allows all of the container strips to be covered by the same lid assembly, although the individual lids 2164 may be removed from the lid assembly 2172. For example, the lid assembly 2172 may include a carrier 2173 to which each lid 2164 is removably attached (e.g., via a weak adhesive, interference fit, or snap - fit mechanism, among others). This configuration allows for easy removal of any single container strip 2118, including its lid 2164, from the other container strips 2118 of the container array 2101 and their lids 2164, for imaging or other processing, etc. In other embodiments, the carrier 2173 may be omitted, and the lids 2164 may be integrally formed with each other and configured to be removable by breaking a frangible connection that joins adjacent pairs of the lids 2164 to each other.

[0162] Figures 36 and 37 show another exemplary inserter 3663 for installation into shell 2162 (see also FIG. 23). Inserter 3663 is the same as inserter 2163, except that inserter 3663 includes a gasket 3674 configured to create a liquid-tight seal with shell 2162. Gasket 3674 may be formed from a softer, more deformable material (e.g., an elastomer), which may be attached to a body 3675 formed from a harder, less deformable material. In some cases, gasket 3674 may be created on body 3675 by overmolding. Gasket 3674 may be located, inter alia, on the lateral sides of body 3675, such as around the lower region of body 3675, or on the bottom side of body 3675.

[0163] Figure 38 shows yet another exemplary inserter 3863 for installation into shell 2162 (see also FIG. 23). Inserter 3863 forms a pair of reservoirs 3805a, 3805b that share side walls with each other instead of being adjacent and separated by a central slot (compare with FIG. 33). Inserter 3863 can be advantageous when a scaffold will be attached to the bottom side of the inserter because there is more surface area available for attachment. Also, inserter 3863 allows the channels at the bottom of the inserter to be more centrally located for vertical alignment with a multicellular structure that is centered within the culture chamber below the inserter.

[0164] Figure 39 shows still another exemplary inserter 3963 for installation into shell 2162 (see also FIG. 23). Inserter 3963 forms four reservoirs 3905a - 3905d and a central slot 3907 located in the middle of the pair of reservoirs (compare with FIG. 33). Inserter 3963 can be advantageous when a multicellular structure is being fed different culture media from the inside and the outside.

[0165] FIG. 40 shows yet another exemplary inserter 4063 for placement into shell 2162 (see also FIG. 23). Inserter 4063 forms four reservoirs 4005a - 4005d, like inserter 3963, but lacks the central slot located between pairs of reservoirs (compare with FIG. 39). Inserter 4063 has more than two reservoirs and combines the potential advantage of placing a channel centrally across a multicellular structure. VI. SELECTED ASPECTS

[0166] This section describes selected aspects of the systems, methods, and devices of the present disclosure as a series of indexed paragraphs.

[0167] Paragraph A1. A system for culturing a multicellular structure such as an organoid, comprising: (a) a container including a culture chamber for containing a multicellular structure (such as an organoid); (b) optionally, an electrical / magnetic module configured to be coupled to and / or located within the container at a position within or adjacent to the culture chamber; and (c) a control circuit configured to wirelessly power and / or operate the electrical / magnetic module, optionally, the electrical / magnetic module is removably coupled to and / or removably located within the container, optionally, the electrical / magnetic module can be received within each of two or more compartments of the container, and optionally, the two or more compartments are selected from a culture chamber, one or more reservoirs, and / or slots.

[0168] Paragraph A2. The system of paragraph A1, wherein the control circuit is configured to wirelessly power and / or operate the electrical / magnetic module using near - field radiation.

[0169] Paragraph A3. The system of paragraph A2, wherein the control circuit is configured to wirelessly transmit power to the electrical / magnetic module via inductive coupling or capacitive coupling.

[0170] Paragraph A4. The control circuit is configured to communicate wirelessly with the electrical / magnetic module via at least one short-range wireless communication protocol, the system according to paragraph A2 or A3.

[0171] Paragraph A5. The electrical / magnetic module is contained within, or configured to be contained within, the culture chamber, includes a magnet, and the control circuit is configured to create a magnetic field that drives the movement of the magnet within the culture chamber, the system according to any of paragraphs A1-A4.

[0172] Paragraph A6. The electrical / magnetic module is a first module, the magnet is a first magnet, and further includes a second module that includes a second magnet, and the first and second modules are contained within, or configured to be contained within, the culture chamber simultaneously, and the control circuit is configured to drive the movement of the first and second magnets relative to each other, optionally, towards and / or away from each other within the culture chamber, the system according to paragraph A5.

[0173] Paragraph A7. The electrical / magnetic module includes a sensor, the system according to any of paragraphs A1-A6.

[0174] Paragraph A8. The electrical / magnetic module includes a chemical sensor, an electrical sensor, an optical sensor, and / or a temperature sensor, the system according to paragraph A7.

[0175] Paragraph A9. The electrical / magnetic module includes electrodes, the system according to any of paragraphs A1-A8.

[0176] Paragraph A10. The electrodes are configured to electrically stimulate and / or electrically sense a multicellular structure within the culture chamber, the system according to paragraph A9.

[0177] Paragraph A11. The electrical / magnetic module includes a light source, the system according to any of paragraphs A1-A10.

[0178] Paragraph A12. The system according to paragraph A11, wherein the light source is configured to illuminate at least a part of the multicellular structure within the culture chamber.

[0179] Paragraph A13. The system according to any one of paragraphs A1 - A12, wherein the electro / magnetic module includes a pump configured to drive fluid flow into and / or out of the culture chamber.

[0180] Paragraph A14. The system according to any one of paragraphs A1 - A13, wherein the electro / magnetic module is located or configured to be located within a slot adjacent to the culture chamber, and optionally, the container defines an opening providing fluid communication between the slot and the culture chamber.

[0181] Paragraph A15. The system according to paragraph A14, wherein the container includes two or more reservoirs in fluid communication with the culture chamber, and optionally, the slot is located vertically above the culture chamber, optionally, between at least a pair of two or more reservoirs, and optionally, each of the two or more reservoirs communicates separately with the culture chamber, and optionally, each of the two or more reservoirs is not connected to any external source of liquid, such as via tubing, and optionally, each of the two or more reservoirs and the culture chamber are formed by and / or located within the same housing, and optionally, each of the two or more reservoirs is directly attached to the culture chamber and / or shares a wall with the culture chamber.

[0182] Paragraph A16. The system according to paragraph A14 or A15, wherein the electro / magnetic module is included within a set of two or more modules configured to perform mutually different functions and coupled to and / or located within the container, and optionally, is interchangeably positioned within the same compartment of the container, such as the same reservoir, culture chamber, and / or slot.

[0183] Paragraph A17. Optionally, each container is located across (optionally, vertically above) the culture chamber, and optionally, each includes at least one or two or more reservoirs that share a wall with the culture chamber, and optionally, two or more reservoirs are formed integrally with each other and / or by the same inserter, and optionally, the electrical / magnetic module is vertically located between the upper side of two or more reservoirs and the bottom of the culture chamber, and / or the system further optionally comprises a removable lid configured to be installed on the container to cover at least one or two or more reservoirs, the system according to any of paragraphs A1 - A16.

[0184] Paragraph A18. Further comprising a container assembly forming a row of containers, the containers of which are interconnected and include a container, and each container of the row of containers includes an individual culture chamber for containing a multicellular structure (such as an organoid, etc.), and optionally, the containers of the row of containers are substantially the same as each other, and optionally, the electrical / magnetic module is movable between the containers of the row of containers, the system according to any of paragraphs A1 - A17.

[0185] Paragraph A19. The container assembly has a length corresponding to the length or width of a standard microplate footprint area, the system according to paragraph A18.

[0186] Paragraph A20. Further comprising a container array including a container assembly and forming at least two or three rows of containers, and each container of at least two or three rows of containers includes an individual culture chamber for containing a multicellular structure, the system according to paragraph A18 or A19.

[0187] Paragraph A21. The system according to any of paragraphs A18 - A20, further comprising a frame for holding a plurality of container assemblies, the frame optionally having a length and / or width corresponding to the length and / or width of a standard microplate footprint, and optionally, the plurality of container assemblies are substantially the same as each other, and optionally, the plurality of container assemblies include corresponding plurality of shells that are substantially the same as each other.

[0188] Paragraph A22. The system according to any of paragraphs A1 - A21, wherein the container includes an optical window formed by the wall of the culture chamber.

[0189] Paragraph A23. The system according to any of paragraphs A1 - A22, further comprising a scaffold positioned or configured to be positioned within the culture chamber and configured to assist in the formation of organoids within the culture chamber.

[0190] Paragraph A24. The system according to any of paragraphs A1 - A23, wherein the container is provided by the device according to any of paragraphs C1 - C20.

[0191] Paragraph B1. A method for culturing a multicellular structure such as an organoid, comprising: (a) a step of containing the multicellular structure in a culture chamber of a container, wherein an electric / magnetic module is removably coupled to and / or located within the container, optionally at a position within or adjacent to the culture chamber; and (b) a step of wirelessly powering / operating the electric / magnetic module using a control circuit. Optionally, the electric / magnetic module is removably coupled to and / or removably located within the container. Optionally, the electric / magnetic module can be received in each of two or more compartments of the container. Optionally, the two or more compartments are selected from a culture chamber, one or more reservoirs, and / or slots. Optionally, the multicellular structure is an organoid with a diameter of at least 0.2, 0.5, 1, or 2 mm. The method is for forming / growing within the culture chamber without connecting the container to an external source of liquid (e.g., a culture medium) (e.g., via tubes and / or pipes) and without electrically connecting the container to a wire or other electrical conductor.

[0192] Paragraph B2. The method according to paragraph B1, wherein the step of powering / operating comprises wirelessly transmitting electrical energy to the electric / magnetic module.

[0193] Paragraph B3. The method according to paragraph B1 or B2, wherein the step of powering / operating is carried out at least in part via inductive or capacitive coupling between the control circuit and the electric / magnetic module.

[0194] Paragraph B4. The method according to any one of paragraphs B1 - B3, wherein the step of powering / operating comprises wirelessly communicating with the electric / magnetic module using near-field radiation.

[0195] Paragraph B5. The method according to paragraph B4, wherein the step of wirelessly communicating comprises exchanging data with the electric / magnetic module using at least one short-range wireless communication protocol.

[0196] Paragraph B6. The electrical / magnetic module includes a magnet, and the step of powering / operating includes driving the movement of the magnet within the culture chamber using a magnetic field created by a control circuit, the method according to any one of paragraphs B1 - B5.

[0197] Paragraph B7. Each culture chamber contains a pair of electrical / magnetic modules each including a magnet, and the step of powering / operating includes driving the electrical / magnetic modules relative to each other, optionally towards and / or away from each other within the culture chamber, the method according to paragraph B6.

[0198] Paragraph B8. The step of powering / operating includes sensing the properties of a multicellular structure (e.g., located within the culture chamber) and / or a culture medium in contact with the multicellular structure using a sensor of the electrical / magnetic module, the method according to any one of paragraphs B1 - B7.

[0199] Paragraph B9. The step of powering / operating includes electrically stimulating the multicellular structure, the method according to any one of paragraphs B1 - B8.

[0200] Paragraph B10. The step of powering / operating includes driving fluid flow into and / or out of the culture chamber, the method according to any one of paragraphs B1 - B9.

[0201] Paragraph B11. The electrical / magnetic module is located within the culture chamber, the method according to any one of paragraphs B1 - B10.

[0202] Paragraph B12. The electrical / magnetic module is located at least predominantly (i.e., more than half by volume ratio) within the container and outside the culture chamber, the method according to any one of paragraphs B1 - B11.

[0203] Paragraph B13. The electrical / magnetic module is located within a slot defined by the container, the method according to paragraph B12.

[0204] Paragraph B14. The method according to any one of paragraphs B1 - B13, further comprising the step of collecting data related to the multicellular structure while the multicellular structure remains within the culture chamber.

[0205] Paragraph B15. The method according to paragraph B14, wherein the step of collecting data includes the step of capturing an image of at least a portion of the multicellular structure.

[0206] Paragraph B16. The method according to paragraph B14 or B15, wherein the step of collecting data is carried out using a sensor of the module.

[0207] Paragraph B17. The method according to any one of paragraphs B1 - B16, further comprising the step of placing an electrical / magnetic module within a compartment of the container.

[0208] Paragraph B18. The method according to paragraph B17, wherein the step of placing includes the step of capturing the electrical / magnetic module within the culture chamber.

[0209] Paragraph B19. The method according to paragraph B17, wherein the step of placing includes the step of optionally installing the electrical / magnetic module within the container but outside the culture chamber, in a reservoir or slot of the container.

[0210] Paragraph B20. Optionally, based on (i) the type of organoid present within and / or to be cultured within the culture chamber, (ii) the culture protocol or protocol stage selected for the organoid, (iii) the test conditions for the organoid, and / or (iv) the parameters to be sensed regarding the organoid and / or the culture medium in contact with the organoid, further comprising the step of selecting the electrical / magnetic module from a set of two or more functionally different modules located outside the container prior to the step of placing. The method according to any one of paragraphs B17 - B19.

[0211] Paragraph B21. The method according to paragraph B20 further includes the step of selecting two or more functionally different modules from the set, and the step of arranging includes, for each selected module, coupling the selected module to the container and / or arranging the selected module within the container.

[0212] Paragraph B22. The set of two or more functionally different modules includes at least one passive module, and optionally, the step of selecting includes the step of selecting a passive module, the method according to paragraph B20 or B21.

[0213] Paragraph B23. The method according to any one of paragraphs B1 - B22, which is implemented using the system according to any one of paragraphs A1 - A24.

[0214] Paragraph C1. A device for culturing a multicellular structure such as an organoid, comprising: (a) optionally, a shell having an open top; and (b) an inserter including at least one reservoir or two or more reservoirs, the inserter being configured to be received within the shell, optionally via the open top, such that the shell and the inserter cooperatively form a culture chamber for the multicellular structure, the culture chamber being located below (optionally, vertically below) two or more reservoirs and being in fluid communication with each of at least one reservoir or two or more reservoirs, optionally via individual channels defined by the inserter.

[0215] Paragraph C2. The device according to paragraph C1, further comprising a lid configured to be installed on the shell to cover each of at least one reservoir and / or two or more reservoirs.

[0216] Paragraph C3. The device according to paragraph C2, wherein the lid is configured to completely cover the open top of the shell.

[0217] The device according to any one of paragraphs C1 or C3, further comprising a scaffold configured to assist in the formation of organoids within the culture chamber.

[0218] The device according to paragraph C4, wherein the scaffold is attached to the wall of the shell.

[0219] The device according to paragraph C4, wherein the shell and the inserter each further comprise a module that provides a scaffold, the module being configured to be (optionally, removably) coupled to and / or located within a compartment of the container, such as a slot defined by the inserter.

[0220] The device according to paragraph C6, wherein the slot is located vertically above the culture chamber and between at least a pair of reservoirs of two or more reservoirs.

[0221] The device according to any one of paragraphs C1 - C7, wherein the shell has an upper region defining a receiving space and a lower region forming a receptacle, the inserter is configured to be received within the receiving space, and the receptacle is configured to cooperate with the inserter to form the culture chamber.

[0222] The device according to paragraph C8, wherein the inserter is configured to form a liquid - tight seal with a plurality of side walls and / or the bottom wall of the upper region of the section.

[0223] The device according to paragraph C9, wherein the inserter includes a body and a gasket attached to the body, the gasket being configured to engage each side wall and / or the bottom wall of the plurality of side walls and form a liquid - tight seal.

[0224] The device according to paragraph C10, wherein the gasket is molded onto the body.

[0225] Paragraph C12. The inserter further comprises a module that forms a slot, defines an opening at the bottom end of the slot, and is configured to be located within or above the slot vertically, optionally, the module is an electrical / magnetic module, a device according to any of paragraphs C1-C11.

[0226] Paragraph C13. The device comprises a plurality of inserters, and the shell forms a column of sections configured to receive the individual inserters of the plurality of inserters respectively, and each section and the individual inserter cooperate to form a culture chamber for a multicellular structure, a device according to any of paragraphs C1-C12.

[0227] Paragraph C14. The sections of the column of sections are integrally formed with each other as a single part, a device according to paragraph C13.

[0228] Paragraph C15. Further comprising a lid configured to be installed on the shell to cover the column of sections, a device according to paragraph C13 or C14.

[0229] Paragraph C16. The lower region of the shell forms at least one optical window of the culture chamber, a device according to any of paragraphs C1-C15.

[0230] Paragraph C17. The lower region of the shell forms at least two optical windows of the culture chamber, a device according to paragraph C16.

[0231] Paragraph C18. The at least two optical windows include a bottom window and at least one side window, a device according to paragraph C17.

[0232] Paragraph C19. The at least two optical windows include a pair of side windows arranged opposite to each other, a device according to paragraph C17 or C18.

[0233] Paragraph C20. The shell and the inserter each form part of the same container and are each configured to be coupled to and / or located within the container, and further comprise a set of two or more functionally different modules, such as one or more modules, optionally at least one or at least two electrical / magnetic modules and / or at least one or at least two passive modules, and optionally at least two of the functionally different modules are configured to be removably installed within the same compartment of the container (e.g., the same reservoir, culture chamber, or slot), the device according to any of paragraphs C1-C19.

[0234] Paragraph D1. A method for culturing a multicellular structure such as an organoid, comprising: (a) optionally, installing an inserter into a shell, the inserter including at least one reservoir or two or more reservoirs, and using the inserter and the shell to cooperatively form a culture chamber, the culture chamber being optionally located (vertically downwards etc.) under each of the at least one reservoir or two or more reservoirs and in fluid communication therewith; and (b) culturing the multicellular structure within the culture chamber.

[0235] Paragraph D2. The method according to paragraph D1, further comprising the step of installing a lid on the shell to cover two or more reservoirs.

[0236] Paragraph D3. The method according to paragraph D1 or D2, wherein the culture chamber is in fluid communication with each of the two or more reservoirs via a channel defined by the inserter.

[0237] Paragraph D4. The method according to any of paragraphs D1-D3, further comprising the step of placing a scaffold within the shell, the scaffold being configured to facilitate organoid formation within the culture chamber, and optionally, cells of the multicellular structure are embedded within the scaffold.

[0238] The step of configuring optionally includes the step of forming a scaffold within the shell before installing the inserter, and is the method described in paragraph D4.

[0239] The step of configuring includes the step of installing a pre-formed scaffold into the shell, and is the method described in paragraph D4.

[0240] The step of installing a pre-formed scaffold into the shell is carried out before installing the inserter, and is the method described in paragraph D6.

[0241] The step of installing a pre-formed scaffold into the shell is carried out by or after installing the inserter. Optionally, the pre-formed scaffold is already attached to the bottom side of the inserter before installing the inserter, and is the method described in paragraph D6.

[0242] The shell includes a column of compartments. The step of installing includes installing at least two inserters into the column of compartments to form two or more separate culture chambers. The step of culturing a multicellular structure includes culturing individual organoids in each of the two or more separate culture chambers, and is the method described in any of paragraphs D1 - D8.

[0243] The method further includes the step of collecting data related to the multicellular structure while the multicellular structure remains within the culture chamber, and is the method described in any of paragraphs D1 - D9.

[0244] The step of collecting data includes the step of capturing an image of at least a part of the multicellular structure, and is the method described in any of paragraphs D1 - D10.

[0245] The method is carried out using any device described in paragraphs C1 - C20, and is the method described in any of paragraphs D1 - D11.

[0246] Paragraph E1. A system for culturing a multicellular structure such as an organoid, comprising: (a) a culture chamber for containing the multicellular structure, and a container including at least one reservoir or at least two or more reservoirs and optional slots, each in fluid communication with the culture chamber; and (b) two or more modules having different functions from each other, configured to be removably coupled to the container and / or disposed therein.

[0247] Paragraph E2. The system according to paragraph E1, wherein the container defines an opening that communicates with the culture chamber at the bottom end of the slot.

[0248] Paragraph E3. The system according to paragraph E2, wherein each of the two or more modules is configured to be positioned adjacent to the opening when installed in the slot.

[0249] Paragraph E4. The system according to any one of paragraphs E1 - E3, wherein the two or more modules include a first module having a permeable membrane.

[0250] Paragraph E5. The system according to paragraph E4, wherein the membrane is configured to form an interface between the liquid in the culture chamber and the liquid or gas in the first module.

[0251] Paragraph E6. The system according to any one of paragraphs E1 - E5, wherein at least one of the two or more modules is an electro / magnetic module.

[0252] Paragraph E7. The system according to paragraph E6, wherein at least one module includes a module having an electronic device.

[0253] Paragraph E8. The system according to paragraph E6 or E7, wherein at least one of the two or more modules does not include an electronic device.

[0254] Paragraph E9. The container is a system according to any of paragraphs E1 - E8, provided by a device according to any of paragraphs C1 - C20.

[0255] Paragraph F1. A method for culturing a multicellular structure such as an organoid, comprising: (a) a step of containing the multicellular structure within a culture chamber of a container, the container defining a slot in communication with the culture chamber and a first module being located within the slot; (b) a step of removing the first module from the slot; and (c) a step of installing a second module into the slot, the second module being configured to perform a function different from that of the first module.

[0256] Paragraph F2. The method according to paragraph F1, wherein the container defines an opening providing communication between the slot and the culture chamber.

[0257] Paragraph F3. The method according to paragraph F1 or F2, wherein at least one of the first and second modules extends from the slot into the culture chamber through the opening.

[0258] Paragraph F4. The method according to any of paragraphs F1 - F3, wherein at least one of the first and second modules includes an electronic device.

[0259] Paragraph F5. The method according to any of paragraphs F1 - F4, wherein the modules of the first and second modules include a membrane forming an interface between the liquid in the culture chamber and the liquid or gas in the module.

[0260] Paragraph F6. The method according to any of paragraphs F1 - F5, wherein at least one of the first and second modules includes an electrode that contacts the multicellular structure and / or the liquid in the culture chamber.

[0261] The method according to any one of paragraphs F1 - F6, further comprising, while one module is located within the slot, using a control circuit to wirelessly power / operate one of the first and second modules.

[0262] The method according to any one of paragraphs F1 - F7, wherein the container is provided by the device according to any one of paragraphs C1 - C20.

[0263] An organoid culture container that does not involve the need to connect tubes and wires to the container itself.

[0264] A method of culturing organoids, comprising the step of forming / growing organoids (e.g., large organoids) within the container without connecting the container to an external source of liquid (e.g., via one or more tubes such as tubes) and / or without electrically connecting the container to an external electrical conductor (e.g., an electrical conductor that is electrically connected to an electrical / magnetic device and / or a control circuit).

[0265] Although the present invention has been described through the above embodiments and features, those skilled in the art will understand that various modifications, combinations, and variations of the embodiments and features can be made without departing from the inventive concept disclosed herein. Also, the present invention should not be regarded as limited to any specific purpose or embodiment described herein, but rather should be regarded as applicable for achieving various purposes other than those described herein. This disclosure has described some embodiments of the technology with reference to the accompanying drawings, which show only some of the possible embodiments. However, other aspects can be embodied in many different forms and should not be construed as limited to the embodiments described herein, even if not explicitly exemplified in combination. Rather, these embodiments are provided so that this disclosure is thorough and complete and fully conveys the scope of the possible embodiments to those skilled in the art.

Claims

1. A device for culturing multicellular structures, such as organoids, comprising: a shell having an open top; an insert including two or more reservoirs, the insert configured to be received within the shell via the open top such that the shell and the insert cooperatively form a culture chamber for the multicellular structure, the culture chamber being located below the two or more reservoirs and in fluid communication with each of the two or more reservoirs via individual channels defined by the insert; A device comprising:

2. The device described in claim 1, further comprising a scaffold configured to support organoid formation within the culture chamber.

3. The device described in claim 2, wherein the scaffolding is provided by a module, the module being positioned within or configured to be positioned within a slot defined by the inserter.

4. A device as described in any of claims 1 to 3, wherein the shell includes a section having an upper region defining a receiving space and a lower region forming a receptacle, the inserter is configured to be received within the receiving space, and the receptacle is configured to cooperatively form the culture chamber with the inserter.

5. A device as described in any one of claims 1 to 4, wherein the inserter forms a slot and defines an opening at a bottom end of the slot, and further comprises a module located or configured to be located within the slot vertically above the opening, and optionally, the module is an electric / magnetic module.

6. A system for culturing a multicellular structure, comprising: a container comprising a culture chamber for containing said multicellular structure and two or more reservoirs and slots, each in fluid communication with said culture chamber; two or more modules having mutually different functions and configured to be interchangeably installed in the slot; A system comprising:

7. The system described in claim 6, wherein the container defines an opening at a bottom end of the slot that communicates with the culture chamber.

8. A system described in any of claims 6 to 7, wherein the two or more modules include a first module having a permeable membrane.

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