Cell culture device

Abstract

Disclosed herein is a cell culture device, a cell culture system, a method of producing a cell culture device, a kit or component, a method for culturing cells, a cell chip, use of such a cell chip, a cell culture device blank, and a cell culture blank system.

Description

Invention Field

[0001] This disclosure pertains to the field of pharmaceutical technology and particularly relates to apparatuses for cell culture and cell assays. The disclosure relates to a cell culture apparatus, a cell culture system, a method for producing a cell culture apparatus, a component kit, a method for culturing cells, a cell chip, uses of such cell chips, a cell culture apparatus blank, and a cell culture blank system. Background Technology

[0002] Biological research on living organisms and attempts to apply this knowledge to constructing in vitro organ models that closely resemble their in vivo counterparts are being conducted in parallel. Organ-on-a-chip (OIA) is an in vitro organ model that holds great potential for improving the efficiency of drug development and reducing reliance on animal testing. To ensure successful integration of OIA into both research and industry, it must closely replicate the physiology and response to treatment of organs in vivo.

[0003] A comprehensive understanding of organogenesis, function, and pathophysiology requires exploring how cells and tissues interact within organs composed of diverse tissue types arranged in a three-dimensional structure with highly dynamic, variable, mechanically complex, and biochemically diverse microenvironments. However, most research in cell and tissue biology relies primarily on 2D cell culture models, which cannot recreate the natural cellular microenvironment present in vivo. Efforts to overcome these limitations have led to the development of 3D cell culture models, in which cells are grown within an extracellular matrix (ECM) that mimics a hydrogel. This approach enhances the expression of differentiation functions and improves tissue arrangement.

[0004] Extensive research over the past few decades has enabled the development of protocols for growing three-dimensional organ-like tissues (called organoids) from tissue-specific stem cells. Organoids exhibit a wide range of histological and functional properties similar to real organs, providing unprecedented opportunities to study tissue development, regeneration, and disease in a laboratory setting without relying on animal testing. However, organoids primarily depend on the spontaneous self-arrangement of stem cells, leading to uncontrolled growth and variability in size and structure. Furthermore, most epithelial stem cell-derived organoids have closed cystic structures, limiting their lifespan and posing challenges for experimental manipulation. Moreover, organoids lack the ability to replicate essential organ features such as tissue-tissue interfaces (e.g., between epithelium and vascular endothelium), spatiotemporal gradients of chemicals and oxygen, and the dynamic microenvironment that plays a crucial role in the functioning of almost all living organs.

[0005] Devices known from the prior art can be divided into membrane-derived devices (which rely on a membrane separating the two sides of the device) and membrane-free devices with hydrogels.

[0006] In membrane-derived devices, the epithelial cell layer is typically grown on a porous membrane inside a cell culture plate. This allows access to the culture medium from both the apical and basal sides of the epithelial layer, making membrane-derived devices a versatile tool for studying in vitro transport, absorption, and other metabolic activities. A common concept is that membrane-based devices consist of two perfusion channels separated by a thin polymer membrane. Human cells can be cultured from both sides of the membrane in these two channels, mimicking the epithelial barrier and physiological functions of living organs such as the lungs, intestines, kidneys, skin, bone marrow, and blood-brain barrier.

[0007] The use of 2D membranes in organ-on-a-chip systems makes it impossible to create 3D tissue structures similar to those in vivo, and makes it impossible to model complex multicellular interactions, such as those arising from non-tissue structures. However, such systems are limiting because they cannot simulate the dynamic microenvironment of living tissue, and the system hinders inherent cell autoarrangement when cells are forced to grow on synthetic 2D membranes. The 2D surface of the membrane flattens cell autoarrangement and impedes interactions between multiple cell types. Modeling various diseases often requires interactions between multiple cell types, including stromal and immune cells. The polymer membranes used in such systems form a physical barrier between epithelial and endothelial cells, preventing direct cell-to-cell interactions. Furthermore, membrane-based systems hinder the introduction of additional cell types, limiting the potential of these cell types to dual-tissue co-culture (e.g., epithelial / endothelial barriers). The only proposed option for multi-tissue culture is to connect various chips via tubing for culture medium exchange, but direct interactions remain impossible. The limited lifespan of approximately 7 to 10 days of culture results in the absence of homeostasis and the inability to model the characteristics of tissue physiology over long periods of health and disease.

[0008] Various limitations of membrane-based systems have spurred the development of first-generation membrane-free hydrogel-based organ-on-a-chip systems. Membrane-free hydrogel organ-on-a-chip platforms exist, allowing the culture of epithelial cells within channels targeting ECM compartments. While these systems are specifically tailored for high-throughput screening of epithelial barrier integrity and absorption, flattening human three-dimensional organs into 2D morphologies impairs in vivo physiological function, reduces cell type composition, and limits the lifespan of these organs. Furthermore, the closed microchannel design of the flat plates makes it difficult to establish gas-liquid interfaces. Three-dimensional cellular structures play a crucial role in organ function, but current technologies still lack a universal microphysiological system that allows for the reproduction of the 3D ECM microenvironment required for proper tissue homeostasis and function. Summary of the Invention

[0009] The overall objective of this disclosure is to advance the prior art in the field of medical technology, particularly in the field of organ-on-a-chip, and preferably to completely or at least partially overcome the disadvantages of the prior art, such as those discussed above. In advantageous embodiments, this disclosure proposes methods for promoting the self-arrangement of cells growing within 3D tissues and for easily analyzing tissues using imaging techniques. Furthermore, multiplex co-culture of cells can be achieved, and complex networks of cells and cell structures can be provided in a simple and reliable manner, providing further understanding of the effects of target compounds on different aspects of different cell types or cell assemblies. In a further advantageous embodiment, this disclosure allows access to multiple different aspects of the hydrogel, i.e., access to the simulated ECM. In advantageous embodiments, a device that can be easily used is provided.

[0010] The overall objective is achieved through the subject matter of the independent claims. Further advantageous embodiments are derived from the dependent claims and the entire disclosure.

[0011] A first aspect of this disclosure relates to a cell culture apparatus. The cell culture apparatus may include a hydrogel compartment containing a hydrogel.

[0012] The cell culture apparatus may further include an inner storage compartment that can be defined or defined by an inner storage wall structure. The inner storage compartment may include a top opening and a bottom opening disposed opposite the top opening. The bottom opening of the inner storage compartment may lead to (e.g., directly to) a hydrogel compartment. Therefore, in some embodiments, the inner storage compartment may be disposed above the hydrogel compartment, particularly directly above it. Thus, the hydrogel may be disposed adjacent to the bottom opening. The top and bottom openings of the inner storage compartment may be configured, and correspondingly arranged, such that the hydrogel in the hydrogel receiving compartment can be accessed from the top opening. Therefore, the hydrogel may be disposed within the hydrogel compartment such that it can be directly accessed from the top opening, for example, via the bottom opening. In some embodiments, the hydrogel may be disposed directly below the bottom opening. That is, the bottom opening is disposed between the hydrogel and the inner storage compartment (and correspondingly, the internal space of the inner storage compartment). In a particular embodiment, the top and bottom openings of the inner storage compartment, and optionally the hydrogel, can be configured and arranged such that the hydrogel in the hydrogel-containing compartment can be accessed along a straight path from the top opening.

[0013] The cell culture apparatus may further include at least one pair, for example, two opposing lateral storage compartments. These opposing lateral storage compartments of the at least one pair may surround an inner storage compartment and, therefore, may also be at least partially defined by the inner storage wall structure. The inner storage wall structure specifically may separate each of the opposing lateral storage compartments from the inner storage compartment and / or may be arranged between each of the opposing lateral storage compartments and the inner storage compartment. While in some embodiments the cell culture apparatus may include only one pair of such opposing lateral storage compartments, in some embodiments it may also include more than one pair of opposing lateral storage compartments, as described herein. With at least two opposing lateral storage compartments and an inner storage compartment, a highly physiological environment for tissue development is provided.

[0014] Because these opposing lateral storage compartments surround the inner storage compartment, the inner storage compartment and these opposing lateral storage compartments can be arranged in a common 2D horizontal plane. This horizontal plane is arranged perpendicular (particularly vertically) to an axis extending through the center of the bottom opening and / or the center of the top opening of the inner storage compartment. In some embodiments, these opposing lateral storage compartments can be arranged at the same height as the inner storage compartment. In certain embodiments, at least one or all of these lateral storage compartments can have a height greater than the height of the hydrogel compartment. In some embodiments, at least one or all of these lateral storage compartments can have a height equal to the sum of the height of the hydrogel compartment and a portion or all of the height of the inner storage compartment. It should be understood that the height of a compartment is its vertical extension. This vertical direction can extend in the direction of the gravity vector and / or along the axis extending (particularly vertically) through the center of the bottom opening and / or the center of the top opening of the inner storage compartment. Because the pair of opposing lateral storage compartments surround the inner storage compartment, a relatively large volume of these lateral storage compartments can be achieved. Furthermore, sufficient gas exchange can be easily performed, which is crucial for cell culture.

[0015] In some embodiments, the cell culture apparatus may include a cell culture apparatus blank, as described in any of the embodiments herein, and, for example, a hydrogel.

[0016] Each of these opposing lateral storage compartments has a height of more than 1000 µm, particularly more than 3000 µm, and even more particularly more than 4000 µm, which is the extension between the bottom section of the lateral storage compartment and the opening.

[0017] In some embodiments, the height of the hydrogel compartment and / or the hydrogel is at least 500 µm, particularly at least 600 µm, and more particularly at least 700 µm. This height of the hydrogel compartment and / or the hydrogel extends along the vertical direction and may be, for example, the distance between the bottom section and the bottom opening of the inner storage compartment.

[0018] In some embodiments, the volume of the hydrogel and / or the hydrogel compartment can be 1 mm. 3 With 500 mm 3 Especially 2 mm 3 With 100 mm 3 between.

[0019] In some embodiments, the contact area of ​​the hydrogel configured to be contacted by culture medium from these opposing lateral storage compartments is more than 40%, particularly more than 50%, more particularly more than 65%, and even more particularly more than 75% of the peripheral surface area of ​​the hydrogel. The peripheral surface area of ​​the hydrogel is the surface area of ​​the peripheral surface (i.e., the surface that extends substantially along the vertical direction and therefore not in the horizontal direction).

[0020] In some embodiments, the hydrogel compartment and / or the hydrogel has an elongated horizontal cross-sectional shape, particularly an elongated circular horizontal cross-sectional shape, such as an ellipse. An elongated shape means a length-to-width ratio greater than 1, particularly greater than 1.5. In some embodiments, the hydrogel compartment and / or the hydrogel has an angled horizontal cross-sectional shape, particularly a polygonal horizontal cross-sectional shape having at least three, particularly at least four, more particularly at least five sides and / or angles. In some embodiments, the polygonal horizontal cross-sectional shape has at most eight, particularly at most six, more particularly at most four sides and / or angles. For example, the hydrogel compartment and / or the hydrogel may have a rectangular horizontal cross-sectional shape. The horizontal cross-section is perpendicular to the vertical direction and may extend, for example, parallel to the bottom section and / or the bottom opening of the inner storage compartment. Polygonal hydrogel compartments or hydrogels are advantageous compared to circular (particularly circular) cross-sectional shapes because circular cross-sectional shapes are inefficient for culture medium diffusion. It has been observed that the diffusion rate through hydrogels with circular horizontal cross-sectional shapes is not high enough to utilize sufficient nutrients from the slow diffusion of fresh culture medium from these opposing lateral storage compartments to support the cells in the center of the hydrogel. Conversely, polygonal, especially triangular, rectangular, or pentagonal horizontal cross-sectional shapes exhibit much higher diffusion rates due to their geometry. Therefore, the cells arranged in the center are provided with sufficient nutrients to allow for ideal and constant cell growth on the hydrogel, such as on its surface.

[0021] In some embodiments, the maximum horizontal width of the hydrogel compartment and / or the hydrogel (i.e., the maximum distance between two opposite peripheral points along a horizontal cross-section) is less than 4 mm, particularly less than 3 mm, and more particularly less than 2.5 mm. In some embodiments, the maximum horizontal width of the hydrogel compartment and / or the hydrogel is at least 0.5 mm, particularly at least 1 mm, and more particularly at least 1.5 mm.

[0022] In some embodiments, the opposing lateral storage compartments in each pair are configured such that they are not directly connected to each other. That is, they do not fuse into each other. In some embodiments, the opposing lateral storage compartments in each pair are configured such that they are indirectly connected to each other only via the hydrogel (and corresponding hydrogel compartment). In other words, the opposing lateral storage compartments in each pair are configured such that culture medium exchange between opposing lateral storage compartments (other than bringing in and out the culture medium through openings in the lateral storage compartments) can only be achieved via the hydrogel or through channels in the hydrogel (and corresponding hydrogel compartment). This separation via the hydrogel allows different culture media to be added to the opposing lateral storage compartments and allows for the study of the diffusion and distribution of these culture media. For example, cells can be added to one storage compartment while a target compound acting as a chemical stimulant is added to the other opposing storage compartment.

[0023] The hydrogel may include a surface that faces and / or is exposed toward the inner storage compartment, particularly via the bottom opening. In some embodiments, the surface may be flat, correspondingly planar, or the surface may include, or in some embodiments may be composed of, morphologies, as described herein in some embodiments. The surface faces and / or is exposed toward the inner storage compartment, particularly via the bottom opening. In other words, when viewed from the top opening toward the bottom opening, it is viewed toward the visible surface.

[0024] In some embodiments, the hydrogel may include a morphology, particularly a micromorphology, having multiple cavities and optionally multiple protrusions. For example, the surface of the hydrogel may include or consist of a morphology, particularly a micromorphology. This micromorphology may face the inner storage compartment and / or may be exposed toward the inner storage compartment, particularly via the bottom opening. In other words, when viewed from the top opening toward the bottom opening, the view is directed toward the morphology, particularly the micromorphology. This morphology, particularly the micromorphology, may be accessible from the inner storage compartment. The described morphology can describe the surface of the hydrogel comprising multiple cavities. Thus, the morphology can be considered an uneven or non-planar surface. This morphology, particularly the micromorphology, allows cells to self-align within 3D tissues. Furthermore, this morphology is very similar to in vivo tissues and their geometry, and further allows for the omission of membranes. In terms of micromorphology, these cavities have an opening from which the cavity begins and extends in the micrometer range, for example, less than 1000 µm, particularly 1 to 1000 µm, and correspondingly less than 1000 µm. 2 Especially 1 to 1000 µm 2 The diameter or open area extends within the range.

[0025] In some embodiments, the hydrogel completely fills the hydrogel compartment.

[0026] This document generally understands that the term "comprising" is interpreted as meaning that it includes the features that follow the term, but it does not exclude the presence of other features, provided they do not invalidate the claim. On the other hand, if the phrase "consisting of..." is used, then there are no further features among the corresponding features besides the features that follow the phrase.

[0027] In some embodiments, the inner compartment may consist only of the top opening and the bottom opening, and therefore may not include any additional openings. For example, the inner storage wall structure may, in particular, laterally and completely surround the inner storage compartment. In some embodiments, the top opening and the bottom opening are arranged above each other and / or may be aligned with each other and / or may be arranged coaxially with each other.

[0028] These opposing lateral storage compartments may be arranged laterally toward both the inner storage compartment and the hydrogel compartment. In some embodiments, the height of each of these opposing lateral storage compartments is equal to the combined height of the hydrogel compartment and the inner storage compartment.

[0029] In some embodiments, these opposing lateral storage compartments and the hydrogel compartment may together form a U-shape in cross-section that preferably surrounds the inner storage compartment.

[0030] In some embodiments, the cell culture apparatus and / or cell culture apparatus preform as described herein do not have an external membrane. For example, there is no membrane between the inner storage compartment and the hydrogel compartment (correspondingly, the hydrogel).

[0031] In some embodiments, the cell culture apparatus includes a bottom section that defines the opposing lateral storage compartments and the hydrogel compartment. In some embodiments, at least a portion of the hydrogel is disposed between the inner storage wall structure and the bottom section. Specifically, a portion of the hydrogel may completely fill the space between the bottom section and the inner storage wall structure. In some embodiments, the bottom section is a plate, such as a common plate forming the bottom sections of the opposing lateral storage compartments and the hydrogel compartment. It is also possible that the bottom section is part of a laboratory dish, such as, for example, a culture dish. The bottom section may be mounted, for example, to the inner storage wall structure and / or the outer wall structure, for example, by a releasable or fixed connection, or they may only be in contact with each other, for example, loose contact, for example, because the inner storage wall structure and / or the outer wall structure rests on the bottom section. It is also possible that the bottom section is not part of the cell culture apparatus. For example, a cell culture apparatus as described herein may form a cell culture assembly together with the bottom section. As an example, a cell culture apparatus can be placed on the bottom section to form the cell culture assembly.

[0032] In some embodiments, the cell culture apparatus includes an outer wall structure. This outer wall structure may form the peripheral lateral boundaries of the cell culture apparatus. In a particular embodiment, the outer wall structure may be arranged such that each of the opposing lateral storage compartments is positioned between the outer wall structure and the inner wall structure.

[0033] In some embodiments, the outer wall structure may contact the bottom section of the cell culture apparatus. Specifically, the bottom section may be connected to the outer wall structure, for example, releasably connected.

[0034] It should be understood that each of these opposing lateral storage compartments (and therefore the lateral active and / or lateral passive storage compartments as well) may include an opening, particularly only one opening, that allows access to the interior of these opposing lateral storage compartments from the external environment. Such an opening can be considered a lateral storage compartment opening. These openings of these opposing lateral storage compartments may be arranged on the same side of the cell culture apparatus as the top opening of the inner storage compartment, particularly at the same height. In some embodiments, the opening of each lateral storage compartment may have a larger open area than the bottom opening of the inner storage compartment. In some embodiments, each lateral storage compartment may be configured such that its bottom section is particularly accessible along a straight path from its opening. Opposing lateral storage compartments including such openings have the advantage that culture medium exchange is facilitated, and can even be automated by robotic pipetting devices. Furthermore, these openings enable highly efficient gas exchange, which is crucial for cell culture, thereby improving cell growth and viability.

[0035] In some embodiments, the open area of ​​the opening in each lateral storage compartment can be 10 mm. 2 With 500 mm 2 Especially 20 mm 2 With 200 mm 2 between.

[0036] The openings described above allow easy access to these lateral storage compartments.

[0037] In some embodiments, the opening of each lateral storage compartment is disposed above the bottom section of the lateral storage compartment, for example, vertically offset from the bottom section of the lateral storage compartment, and particularly only vertically offset. Specifically, the opening of each lateral storage compartment is configured such that at least 50%, particularly at least 70%, more particularly at least 90%, and even more particularly at least 95% of the bottom section of each lateral storage compartment is uncovered. This means that no wall portion is vertically disposed above these uncovered portions of the bottom section, and / or the open area defined by the opening of the lateral storage compartment is vertically disposed above these uncovered portions of the bottom section.

[0038] In some embodiments, the openings of these lateral storage compartments are arranged parallel to and / or coaxial with the bottom section (of the lateral storage compartment).

[0039] In some embodiments, at least some or even all of the openings of these lateral storage compartments and / or the top openings of the inner storage compartments are uncovered and open to the environment.

[0040] In some embodiments, the opposing lateral storage compartments have a cylindrical shape. Such a cylindrical shape can be regular or irregular. A regular cylinder has a regular bottom surface, such as circular (circular cylinder) or rectangular (prism), or an irregular shape, and has a cylindrical shell extending particularly uniformly and / or linearly between the top and bottom surfaces. In a particular embodiment, the top surface of the cylindrical shape is the opening of the corresponding lateral storage compartment, and the bottom surface is the bottom section of the corresponding lateral storage compartment.

[0041] In some embodiments, the top opening and the bottom opening, as well as the openings of these optional lateral storage compartments, can be considered accordingly as 2D planes.

[0042] In some embodiments, the open area defined by the top opening is equal to or greater than the open area defined by the bottom opening.

[0043] In some embodiments, the top opening and / or the bottom opening may be circular, particularly circular. In alternative embodiments, the top opening and the bottom opening may be angled.

[0044] In some embodiments, the hydrogel is disposed within the hydrogel compartments such that the opposing lateral storage compartments and the inner storage compartment are indirectly fluidly connected to each other only via the hydrogel (i.e., through the hydrogel itself or through channels protruding from the hydrogel). In some embodiments, the hydrogel may separate the inner storage compartment from the opposing lateral storage compartments. As used herein, the term fluid connection may specifically refer to liquid fluid connection in some embodiments.

[0045] In some embodiments, the hydrogel completely fills the hydrogel compartment (optionally, except for the channels protruding through the hydrogel).

[0046] In some embodiments, the inner reservoir wall structure includes an inward protrusion. In some embodiments, the inward protrusion particularly peripherally and completely defines the bottom opening. In some embodiments, the inward protrusion is disposed between the inner reservoir compartment and the hydrogel. In a particular embodiment, the hydrogel contacts the inward protrusion. The inward protrusion is advantageous because it holds the hydrogel in place, particularly in case of undesirable removal of the hydrogel toward the inner reservoir compartment. Thus, the inward protrusion retains the hydrogel in the hydrogel compartment. This is particularly advantageous if a casting insert as described herein is used in some embodiments of the method of manufacturing a cell culture device. The inward protrusion helps to hold the hydrogel in place when the casting insert is removed. Furthermore, the inward protrusion can serve as a resting surface on which the casting insert can be rested during the manufacturing of the cell culture device as described herein. The inner protrusion can improve the quality of device production and the device itself because it ensures that the hydrogel has a constant thickness and that the hydrogel of multiple devices produced by the same method is constant and renewable.

[0047] In some embodiments, the inner storage wall structure includes a vertical inner wall section. This vertical inner wall section typically extends perpendicular to a 2D plane defined by the top opening and / or the bottom opening. The vertical inner wall section may have an inner surface facing and correspondingly defining the inner storage compartment. The vertical inner wall section may also have one or more outer surfaces, each facing and correspondingly defining at least one of these opposing lateral storage compartments.

[0048] For example, the inner protrusion may be arranged adjacent to the vertical inner wall section and at an angle to it, particularly at an angle of <180° to 90°, particularly 150° to 90°.

[0049] In some embodiments, the inner storage wall structure includes a vertical inner wall section and an inclined inner wall section arranged adjacent thereto. The vertical and inclined inner wall sections may at least partially define at least one of the opposing lateral storage compartments. Alternatively, each lateral storage compartment may be at least partially defined by such vertical and inclined inner wall sections. Such inclined inner wall sections have the advantage that air present in the corresponding lateral storage compartment can be easily vented without being trapped within that compartment. In some embodiments, the inner storage wall section may not contain a vertical inner wall section, but may still include an inclined inner wall section inclined toward the vertical direction of the cell culture apparatus. As noted herein, this vertical direction may be aligned with the gravity vector and / or may be perpendicular to the top opening of the inner storage compartment. For example, the inclined inner wall section can be arranged in cross-section at an angle of 10° to 80°, particularly 25° to 60°, and more particularly 30° to 50° to the vertical direction.

[0050] In some embodiments, the cell culture apparatus may also include one or more venting channels. For example, at least one or all of these lateral storage compartments may be connected to such venting channels. For example, the opening of the venting channel may open to these lateral storage compartments. Such venting channels may be arranged such that any remaining gas in the lateral storage compartment is discharged from the lateral storage compartment through the venting channel during filling (e.g., using liquid culture medium). Thus, the venting channel may be open to the external environment.

[0051] In some general embodiments, these opposing lateral storage compartments can be configured such that any residual gas in these lateral storage compartments is vented from them during filling (e.g., using a liquid culture medium). This avoids air trapping in these lateral storage compartments.

[0052] In some embodiments, the inclined inner wall section may form an interior angle (i.e., an angle surrounding a portion of the inner storage wall structure), the interior angle being between 90° and 180°, particularly between >90° and <180°, such as 100° to 170°, or 110° to 160°, or 120° to 150°.

[0053] In some embodiments, the inclined inner wall section is configured such that the cross-sectional open area of ​​at least one lateral storage compartment at least partially defined by the inclined inner wall section decreases toward the hydrogel compartment. This means that the cross-section (i.e., the plane parallel to the vertical direction) decreases toward the hydrogel compartment (and correspondingly the hydrogel), particularly continuously.

[0054] For example, the inclined inner wall section may have a conical or truncated conical shape, such as a partially conical or partially truncated conical shape. It should be understood that the tip of the cone or truncated cone points towards the hydrogel compartment, and correspondingly, the hydrogel. This provides a conical or truncated conical path from the lateral storage compartment toward the hydrogel compartment. Therefore, in some embodiments, at least one or all of these opposing lateral storage compartments may include a conical or truncated conical path or a partially conical or partially truncated conical path toward the hydrogel compartment. A partially conical shape is a cross-section of a cone, particularly a cross-section along a plane parallel to or even along the axis of rotation of the cone. The same applies to partially truncated conical shapes.

[0055] In some embodiments, the inclined inner wall section is disposed between the vertical inner wall section and the inner protrusion. The inclined inner wall section may in particular be inclined toward the inner protrusion and the vertical inner wall section. In a particular embodiment, the inner protrusion may be at an angle relative to the vertical inner wall section, for example, an angle of 80° to 100°, particularly 85° to 95°, and more particularly substantially 90°.

[0056] In some embodiments, the inner reservoir wall structure includes one or more support structures that laterally surround the hydrogel or a portion thereof. These support structures may be, for example, multiple struts. In some embodiments, these support structures may project from the inner protrusion specifically toward the bottom section. In some embodiments, these support structures may extend upward toward and thus contact the bottom section. In other embodiments, these support structures may extend toward the bottom section but not upward, thus forming a gap between themselves and the bottom section. In some embodiments, the one or more support structures may have a height (i.e., along the vertical direction, correspondingly extending from the inner reservoir wall structure toward the bottom section) that is 90% or less of the height of the hydrogel compartment and / or the hydrogel.

[0057] In some embodiments, the inner storage wall structure includes a plurality of support structures. These support structures may be spaced apart from each other and define one or more gaps between them. In some embodiments, the gap between adjacent support structures may be at least 700 µm, and particularly more than 800 µm. In some embodiments, the one or more support structures are arranged around the bottom opening. These support structures may also be spaced apart from the bottom opening. The described support structures are advantageous because they provide further securing to the hydrogel and help maintain the hydrogel in place, particularly against lateral displacement toward these lateral storage compartments. This is particularly advantageous during the filling of the hydrogel precursor into the hydrogel compartment, as it allows the hydrogel precursor to be held inside the hydrogel compartment. In some embodiments, the one or more support structures are arranged radially around the bottom opening.

[0058] In some embodiments, the inner reservoir wall structure surrounds the periphery of the hydrogel. In some embodiments, at least 50%, particularly at least 65%, more particularly at least 75%, and even more particularly at least 80% of the peripheral surface area of ​​the hydrogel is exposed and therefore not covered by the one or more support structures (i.e., uncovered). Such embodiments are advantageous because while the hydrogel is efficiently held in its position, a relatively large surface area is still available for culture medium and compound diffusion. The peripheral surface area is the surface area of ​​the peripheral surface (i.e., the surface that extends substantially along the vertical direction and therefore not in the horizontal direction).

[0059] In some embodiments, the one or more support structures may define the lateral (i.e. peripheral) boundaries of the hydrogel compartment.

[0060] In embodiments where the cell culture apparatus includes a hydrogel delivery orifice (as further described below), the support structures are preferably arranged such that they surround the hydrogel delivery orifice, which in turn leads into the delivery outlet of the hydrogel compartment.

[0061] In some embodiments, the inner storage wall structure separates the opposing lateral storage compartments from the inner storage compartment, specifically such that the opposing lateral storage compartments and the inner storage compartment are fluidly connected only indirectly via the hydrogel compartment and / or the hydrogel.

[0062] In some embodiments, the hydrogel compartment and / or the hydrogel is arranged between these opposing lateral storage compartments.

[0063] In some embodiments, the hydrogel compartment and / or the hydrogel may be surrounded by and / or fluidly connected to the inner storage compartment and these opposing lateral storage compartments.

[0064] In some embodiments, the morphology, particularly the micromorphology, is configured such that the cavities are not undercut. In some embodiments, these cavities may have a dome or spherical cross-section or a cylindrical, particularly circular, cylindrical shape.

[0065] In some embodiments, each cavity may have a cavity depth of 1 to 1000 µm. In some embodiments, each cavity may have a depth of 1 to 100000 µm. 3 The volume of the cavity.

[0066] In some embodiments, each cavity may have a cavity opening, i.e., an opening extending from the cavity therefrom, the cavity opening defining a depth of 1 to 100,000 µm. 2 The open area.

[0067] In some embodiments, the morphology, particularly the micromorphology, includes at least one cavity, particularly at least 10 cavities, particularly at least 20 cavities, and more particularly at least 50 cavities.

[0068] In some embodiments, the cell culture apparatus further includes a hydrogel delivery orifice opening toward the hydrogel compartment. The hydrogel delivery orifice may be configured to deliver hydrogel or a hydrogel precursor into the hydrogel compartment. In some embodiments, the hydrogel delivery orifice includes a delivery outlet directly leading to the hydrogel compartment. In some embodiments, the hydrogel delivery orifice has an orifice inlet through which the hydrogel or hydrogel precursor can be filled. In some embodiments, the orifice inlet is arranged adjacent to the top opening of the inner storage compartment and / or on the same side of the cell culture apparatus blank as the top opening of the inner storage compartment. In some embodiments, the orifice inlet may be offset from the top opening, particularly laterally offset. In some embodiments, the orifice inlet is defined by the inner storage wall structure.

[0069] In some embodiments, the cell culture apparatus further includes a hydrogel buffer compartment. The hydrogel buffer compartment may be configured to receive hydrogel and / or hydrogel precursors during filling. The hydrogel compartment may have access to (particularly direct access to) the hydrogel compartment. For example, the hydrogel buffer compartment may include a hydrogel buffer compartment opening via which the hydrogel buffer compartment is connected to the hydrogel compartment. In a particular embodiment, the hydrogel buffer compartment is separated from the hydrogel delivery orifice. For example, the hydrogel buffer compartment may not be directly connected to the hydrogel delivery orifice, but both the hydrogel delivery orifice and the hydrogel buffer compartment have access to the hydrogel compartment. In some embodiments, the hydrogel buffer compartment may include a vent opening configured to vent air from the hydrogel buffer compartment to the external environment during filling of the hydrogel compartment, and consequently the hydrogel buffer compartment. Typically, the vent opening is different from the hydrogel buffer compartment opening. In some embodiments, the hydrogel buffer compartment may be arranged opposite to the hydrogel delivery orifice, and / or the delivery outlet of the hydrogel delivery orifice may be arranged opposite to the opening of the hydrogel buffer compartment. In some embodiments, the hydrogel buffer compartment may have a volume of 1 to 100 µL, particularly 2 to 20 µL.

[0070] In some specific embodiments, the cell culture apparatus may include:

[0071] - Hydrogel compartment, which contains hydrogel;

[0072] - An inner storage compartment defined by an inner storage wall structure, the inner storage compartment including a top opening and an opposing bottom opening, wherein the bottom opening leads to the hydrogel compartment, and wherein the top opening and the bottom opening are configured such that the hydrogel can be accessed from the top opening, wherein the inner storage wall structure includes an inner protrusion that defines the bottom opening and / or is disposed between the inner storage compartment and the hydrogel, wherein preferably, the hydrogel (3) contacts the inner protrusion (14);

[0073] - At least one pair of opposing lateral storage compartments, which surround the inner storage compartment and each leads to the hydrogel compartment, such that the hydrogel is in fluid communication with each lateral storage compartment;

[0074] - A hydrogel delivery orifice, which is open toward the hydrogel compartment and configured to deliver a hydrogel or hydrogel precursor into the hydrogel compartment;

[0075] The hydrogel includes a surface that faces and / or is exposed toward the inner storage compartment, particularly through the bottom opening.

[0076] In some embodiments, at least one of a pair of opposing lateral storage compartments is a lateral passive storage compartment. This means, for example, that active transport of culture medium or compounds through these lateral passive storage compartments through the hydrogel may not exist, but such culture medium or compounds can only diffuse passively through the hydrogel. Therefore, the hydrogel can be configured, for example, such that transport of culture medium and / or compounds through the hydrogel from any of the lateral passive storage compartments can be controlled solely by the diffusion rate of the culture medium and / or compounds through the hydrogel. In some embodiments, the hydrogel may not have channels leading to any of the lateral passive storage compartments, such that culture medium or any compound can only be exchanged between the lateral passive storage compartments by diffusion through the hydrogel (other than bringing in and out culture medium via openings in the lateral passive storage compartments). Since the hydrogel simulates the ECM, this can be an advantageous model system for investigating the effect of one compound on the diffusion behavior of other compounds through the ECM (i.e., the hydrogel). In these embodiments, the cell culture apparatus includes at least one pair or optionally only one pair of opposing lateral storage compartments, in which case these lateral storage compartments are lateral passive storage compartments.

[0077] In some embodiments, at least one of a pair of opposing lateral storage compartments is a lateral active storage compartment. In contrast to lateral passive storage compartments, active transport through the hydrogel can be achieved, for example, by means of channels protruding through the hydrogel. Such channels can, for example, connect two lateral active storage compartments to each other. Alternatively or additionally, one or more channels may connect one or both of the lateral active storage compartments to an inner storage compartment. Thus, such channels typically protrude through the hydrogel and lead to at least two storage compartments (i.e., storage compartments selected from the two lateral active storage compartments and the inner storage compartment). In some embodiments, the hydrogel defines one or more channels that extend between and lead to each of the two lateral active storage compartments. These channels are preferably configured for active culture medium exchange between the lateral active storage compartments. Active culture medium exchange, in contrast to diffusion-controlled passive culture medium exchange, refers to the flow of culture medium between at least two storage compartments connected by such channels. In some embodiments, a pump unit may even be present, which can be configured to actively pump culture medium through the one or more channels. Gravity-driven flow can also be induced through these channels, for example, by tilting the cell culture apparatus or by providing tilted channels. In the described embodiments, the cell culture apparatus includes at least one pair, or optionally only one pair, of opposing lateral storage compartments, in which case these lateral storage compartments are laterally active storage compartments.

[0078] In some embodiments, there may also be more than one pair of opposing lateral storage compartments. Typically, two opposing lateral storage compartments form a storage compartment pair. In some embodiments, for example, four lateral storage compartments may be present, i.e., two pairs of opposing lateral storage compartments, each consisting of two opposing lateral storage compartments. In some embodiments, it is possible that the cell culture device has one pair of opposing lateral active storage compartments and one pair of opposing lateral passive storage compartments. Thus, in such embodiments, the hydrogel simulating an ECM can be accessed from five sides, i.e., from the inner storage compartment, the two lateral active storage compartments, and the two lateral passive storage compartments. However, it is also possible that the cell culture device has a first pair of opposing lateral passive storage compartments and a second pair of opposing lateral passive storage compartments. It is also possible that the cell culture device has a first pair of opposing lateral active storage compartments and a second pair of opposing lateral active storage compartments.

[0079] In some embodiments, the one or more channels defined by the hydrogel may have a diameter of 10 µm. 2 With 1mm 2 Specifically, 20 µm 2 With 0.8 mm 2 The open area of ​​the cross-section between them.

[0080] In some embodiments, the one or more channels may originate from a channel network. In a particular embodiment, the channel network may include one or more of channel intersections, recesses, protrusions, and / or branches. Channels having, for example, recesses and / or protrusions can be used to simulate certain internal organ structures. The channel network can, for example, simulate networks of microvessels and lymphatic vessels, and can, for example, allow the observation of compound migration or fluid flow through these vessels.

[0081] In some embodiments, the inner storage compartment may have a volume of 0.01 to 10 mL, particularly 0.05 to 0.5 mL. In some embodiments, these opposing lateral storage compartments may each have a volume of 0.01 to 10 mL, particularly 0.05 to 0.5 mL. Such larger volume capacities are advantageous because they allow for the long-term use of the culture medium present in these storage compartments without depletion, and thus reduce the frequency of required culture medium exchange operations.

[0082] In some embodiments, the hydrogel comprises or is composed of the following: selected from polysaccharides, gel proteins, agarose, alginate, chitosan, dextran, gelatin, laminin, collagen, hyaluronic acid, and fibroin; cross-linked synthetic hydrophilic polymers, particularly functionalized using extracellular matrix (ECM)-derived proteins or peptides; and mixtures thereof. In some embodiments, the cross-linked synthetic hydrophilic polymer may be selected from the group consisting of: poly(ethylene glycol), polyoxazoline, polyaliphatic polyurethane, polyether polyurethane, polyester polyurethane, polyethylene copolymer, polyamide, polyvinyl alcohol, poly(ethylene oxide), polypropylene oxide, polypropylene glycol, polytetramethylene oxide, polyvinylpyrrolidone, polyacrylamide, poly(hydroxyethyl acrylate), poly(hydroxyethyl methacrylate), and compounds or copolymers thereof. In some embodiments, the hydrogel comprises a basement membrane-like matrix (Matrigel) or a human extracellular matrix (such as Myogel).

[0083] A second aspect of this disclosure relates to a cell culture system. Such a cell culture system may include a plurality of cell culture devices as specifically described with respect to the first aspect in any of the embodiments herein. For example, such a cell culture system may include at least 2, at least 4, at least 10, at least 20, at least 50, at least 90, or at least 100 cell culture devices. In a particular embodiment, the cell culture system may be configured as a well plate. Such a well plate may include all cell culture devices and allow for several screenings to be performed in parallel under different conditions. In particular, these cell culture devices may be arranged in a matrix of rows and columns.

[0084] A third aspect of this disclosure relates to a method for producing a cell culture apparatus, specifically as described in any of the embodiments herein (such as the embodiments of the first aspect).

[0085] The method may include the step of providing a cell culture device blank. This cell culture device blank may specifically be a cell culture device blank as described in any of the embodiments herein. It should be understood that the cell culture device blank differs from the cell culture device in that the cell culture device blank does not yet contain hydrogel inside the hydrogel compartment. However, in some embodiments, this may be the only difference between the cell culture device and the cell culture device blank. Therefore, any of the embodiments described herein with respect to the cell culture device may also be applied to this cell culture device blank. The cell culture device blank may include a hydrogel compartment, an inner storage compartment defined by an inner storage wall structure, and a pair of opposing lateral storage compartments surrounding the inner storage compartment and each opening to the hydrogel compartment. The inner storage compartment may include a top opening and opposing bottom openings. The bottom opening can lead to the hydrogel compartment, and both the top opening and the bottom opening (and optionally the inner storage compartment) can be configured such that the hydrogel compartment can be accessed from the top opening, particularly along a straight path from the top opening to the bottom opening.

[0086] The method may further include the step of introducing a casting insert through the top opening into the inner storage compartment. The casting insert may be introduced such that it covers the bottom opening, particularly completely covering the bottom opening. In some embodiments, the casting insert may include a casting surface. In a particular embodiment, the casting insert may be introduced into the inner storage compartment such that the casting surface of the casting insert is exposed toward (correspondingly, facing) the hydrogel compartment.

[0087] In certain embodiments, the cast surface of the cast insert may include a morphology, particularly a micromorphology, having multiple protrusions and optional cavities. In some embodiments, the cast insert may be introduced into the inner storage compartment such that the morphology, particularly the micromorphology, of the cast insert is exposed toward the hydrogel compartment (correspondingly facing the hydrogel compartment or even protruding therein). The cast insert may, for example, protrude at least partially through the bottom opening.

[0088] The cast insert may specifically include: a base portion including the cast surface, particularly the morphology, and correspondingly the micromorphology; and a rod, particularly a straight rod, mounted on the side of the base portion opposite the cast surface. For example, the cast insert can be considered a stamped part. The cast surface may correspond to the surface of the hydrogel, which faces and / or is exposed toward the inner storage compartment, particularly via the bottom opening. In other words, the cast surface of the cast insert acts as a negative for the surface of the hydrogel.

[0089] The method may further include the step of filling the hydrogel compartment with a hydrogel precursor. Specifically, the hydrogel precursor may be filled into the hydrogel compartment such that the hydrogel precursor contacts the casting insert, particularly the casting surface of the casting insert. Thus, upon curing, a surface of the hydrogel corresponding to the casting surface is formed.

[0090] The method may further include the step of curing the hydrogel precursor to form a hydrogel, the hydrogel including a surface facing and / or exposed toward the inner storage compartment, particularly via the bottom opening. This surface of the hydrogel may correspond to the casting surface of the casting insert.

[0091] The method may further include the step of removing the cast insert through the top opening. Thus, the cell culture apparatus can be provided.

[0092] In some embodiments, the casting surface may include or consist of a morphology, particularly a micromorphology, having a plurality of protrusions. This morphology (corresponding micromorphology) of the casting insert corresponds to the morphology (corresponding micromorphology) of the hydrogel. In other words, the morphology (corresponding micromorphology) of the casting insert acts as a negative for the morphology (corresponding micromorphology) of the hydrogel. Therefore, each protrusion of the morphology (corresponding micromorphology) of the casting insert creates a cavity of the morphology (corresponding micromorphology) of the hydrogel, and / or any cavity of the morphology (corresponding micromorphology) of the casting insert creates a protrusion of the morphology (corresponding micromorphology) of the hydrogel.

[0093] In some embodiments, the cast insert may be introduced into the inner storage compartment through the top opening, such that the cast insert covers the bottom opening and that the morphology (particularly the micromorphology) of the cast insert is exposed to and / or protrudes into the hydrogel compartment. When the hydrogel precursor is filled into the hydrogel compartment, the hydrogel precursor surrounds these protrusions of the micromorphology of the cast insert. Curing of the hydrogel precursor can subsequently form a hydrogel comprising a micromorphology with multiple cavities.

[0094] In some general embodiments, curing may include chemical conversion. For example, curing may include hardening and / or polymerization. For example, curing may include or consist of light treatment, such as UV irradiation, heating and / or cooling.

[0095] In some embodiments, the inner storage wall structure of the cell culture apparatus blank includes an inner protrusion. This inner protrusion may be (i.e., the same as described with respect to a corresponding embodiment of the cell culture apparatus as described in the first aspect of this disclosure) an inner protrusion. The inner protrusion may define the bottom opening and / or may be disposed between the inner storage compartment and the hydrogel compartment. In a particular embodiment, the inner protrusion may form the peripheral boundary of the bottom opening.

[0096] In some embodiments, the casting insert is introduced into the inner storage compartment such that it rests on the inner protrusion of the inner storage wall structure. Specifically, the casting insert may be introduced to seal the bottom opening. It should be understood that the casting insert and the cell culture apparatus blank are preferably configured such that the casting insert rests on the inner protrusion and completely covers the bottom opening. Furthermore, the casting insert and the cell culture apparatus may be configured such that when the casting insert rests on the inner protrusion, the casting surface (particularly the morphology or micromorphology) of the casting insert faces and / or protrudes into the hydrogel compartment.

[0097] In some embodiments, the hydrogel precursor may be filled into the hydrogel compartment in such a manner that the central portion of the hydrogel precursor is positioned below the casting insert and the bottom opening after being filled into the hydrogel compartment, and the peripheral portion of the hydrogel precursor is positioned below the inner protrusion of the inner storage wall structure. In a particular embodiment, the hydrogel precursor may be filled into the hydrogel compartment in such a manner that the inner protrusion is positioned between the hydrogel precursor (particularly the peripheral portion of the hydrogel precursor) and the inner storage compartment. The terms "peripheral portion" and "central portion" refer to the regions of the hydrogel precursor after it has been filled into the hydrogel compartment. The peripheral portion surrounds, particularly circumferentially, the central portion along its lateral periphery. The peripheral portion and the central portion may have a cylindrical shape. The central portion may typically be positioned directly below the bottom opening (correspondingly the casting insert and its morphology), and the peripheral portion may be positioned below the inner protrusion.

[0098] In some embodiments, one or more support structures protrude from the inner protrusion. These support structures may be, for example, the same as those described herein with respect to the first aspect of this disclosure. During the filling of the hydrogel precursor into the hydrogel compartment, the one or more support structures may retain the hydrogel precursor within the hydrogel compartment.

[0099] In some embodiments, the cell culture apparatus preform includes a hydrogel delivery orifice opening toward the hydrogel compartment. This hydrogel delivery orifice may be, for example, the same as described herein with respect to the first and / or eleventh aspects of this disclosure. The hydrogel precursor can be supplied into the hydrogel compartment during filling via the hydrogel delivery orifice. In particular, the hydrogel delivery orifice may have an orifice inlet from which the hydrogel precursor can be supplied directly into the hydrogel compartment, for example, via a delivery orifice outlet of the hydrogel delivery orifice. The delivery orifice outlet may be surrounded by support structures as described in the embodiments herein. This ensures that the hydrogel precursor is introduced centrally and flows from there toward these support structures, which then prevent further flow of the hydrogel precursor, particularly into these opposing lateral storage compartments.

[0100] In some embodiments, the opposing lateral storage compartments in the pair of opposing lateral storage compartments are lateral active storage compartments as described, for example, with respect to the embodiments of the first aspect. One or more channels may be formed that extend through the cured hydrogel and lead to each of the two lateral active storage compartments. The formation of the one or more channels may be performed after the hydrogel precursor has been cured. Specifically, this may be achieved, for example, by depleting or degrading portions of the cured hydrogel. Suitable methods may be, for example, photodegradation (such as laser ablation or photoinduced depolymerization) or chemical degradation. Alternatively, the channels may have already been formed, for example, during the filling of the hydrogel precursor using suitable mold inserts. For example, rods such as microrods may be introduced into the cell culture device preform before the hydrogel precursor is filled into the hydrogel compartments. These rods may then be removed after curing to create the channels.

[0101] In some embodiments, the hydrogel precursor includes cells to be cultured (i.e., prior to the hydrogel precursor being filled into the hydrogel compartment). This has the advantage that the behavior of the cells can be monitored within the hydrogel under certain conditions. For example, the diffusion, growth, and / or release of signaling substances from these cells into the hydrogel can be monitored. These cells can be suspended in the hydrogel precursor, for example, before the hydrogel precursor is filled into the hydrogel compartment.

[0102] Generally, the cells used in this article can be living cells. These cells may be derived from human or animal donors, for example. In some embodiments, these cells may be stem cells.

[0103] A fourth aspect of this disclosure relates to a component kit. This component kit can be used, for example, in any of the methods described in the embodiments of the third aspect of this disclosure.

[0104] The kit may include a cell culture apparatus blank, specifically, a cell culture apparatus blank as described herein, such as in embodiments relating to the third or eleventh aspect. The kit may alternatively include a cell culture blank system, specifically, a cell culture blank system as described herein, such as in embodiments relating to the twelfth aspect.

[0105] The cell culture apparatus prototype may include a hydrogel compartment and an inner storage compartment defined by an inner storage wall structure. Furthermore, the cell culture apparatus prototype may include at least a pair of opposing lateral storage compartments that surround the inner storage compartment and each open to the hydrogel compartment. The inner storage compartment may include a top opening and opposing bottom openings. The bottom opening may open to the hydrogel compartment. Moreover, the top opening and the bottom opening may be configured such that the hydrogel compartment is accessible from the top opening, particularly along a straight path from the top opening.

[0106] In some embodiments, the kit may further include a casting insert. The casting insert may be as described herein, for example, with respect to a third aspect of this disclosure. The casting insert may include a casting surface. The casting insert may be further configured to be introduced into the inner storage compartment through the top opening, such that the casting insert covers the bottom opening. The casting insert and the cell culture apparatus preform may be further configured such that the casting surface morphology (particularly micromorphology) is exposed to the hydrogel compartment.

[0107] The kit may optionally also contain a hydrogel precursor.

[0108] In some embodiments, the casting surface includes or is composed of a morphology having a plurality of protrusions, particularly a micromorphology. The casting insert may be configured to be introduced into the inner storage compartment through the top opening such that the casting insert covers the bottom opening and such that the morphology (particularly the micromorphology) is exposed to and / or protrudes into the hydrogel compartment.

[0109] The fifth aspect of this disclosure relates to a method for culturing cells using a cell culture apparatus, particularly using a cell culture apparatus as described herein, for example, with respect to embodiments of the first aspect. This method can also be performed using a cell culture system as described herein, particularly with respect to the second aspect.

[0110] The method may include the step of specifically providing a cell suspension comprising cells and liquid culture medium into the inner storage compartment via the top opening.

[0111] The method may further include the steps of allowing these cells to deposit on the surface of the hydrogel, particularly into the plurality of cavities of the hydrogel's morphology (especially micromorphology), and culturing these cells. Sedimentation may be performed for a specific sedimentation time interval. This sedimentation time interval may be, for example, 1 to 15 minutes, particularly 5 to 10 minutes. In some embodiments, the method may include centrifuging the cell culture apparatus. Centrifugation may include the sedimentation of these cells. By using such a centrifugation step, sedimentation can be accelerated.

[0112] This method may further include culturing the sedimented cells.

[0113] In some instances, the method according to the fifth aspect can be analogous to forming a cellular barrier, such as an epithelial cell barrier, between the apical side (e.g., the top reservoir) and the basal side (e.g., the hydrogel and optionally these opposing lateral reservoir compartments). Therefore, targeted exposure to a target compound (e.g., a drug) can be assessed independently on both the basal and apical sides.

[0114] In some embodiments, a majority (i.e., at least 50 vol.%, for example, at least 80 vol.%, for example, at least 90 vol.%, for example, at least 95 vol.%) or all of the liquid culture medium of the cell suspension is removed from the inner storage compartment, particularly via the top opening, after the cells have settled. This can be done, for example, by a suitable cannula or syringe.

[0115] After the liquid culture medium has been removed, different culture media (e.g., cell culture medium, preferably liquid cell culture medium) can be added to the inner storage compartment and to the surface of the hydrogel (e.g., morphology, correspondingly micromorphology) and the settled cells.

[0116] Alternatively, after the liquid culture medium has been removed, a hydrogel precursor can be filled into an inner storage compartment covering the settled cells. The hydrogel precursor can then be solidified, thereby forming a hydrogel. This hydrogel can be considered an inner storage hydrogel because it is present within the inner storage compartment. The hydrogel precursor can be the same as that used in embodiments of the third aspect (e.g., the production of cell culture apparatus), or it can be different. Thus, in some embodiments, the inner storage hydrogel can be made of the same material as the hydrogel in the hydrogel compartment, or it can be made of a different material. Therefore, the inner storage hydrogel can (together with the hydrogel in the hydrogel compartment) completely enclose the settled cells. This allows for the creation of 3D tissue cultures.

[0117] In some embodiments, one or more of the opposing lateral storage compartments may be filled with culture medium. The culture medium may be the same for all opposing lateral storage compartments, or it may be different. It is also possible that at least one of the lateral storage compartments is not filled with culture medium, i.e., the lateral storage compartment is empty and may contain only air. As used herein, the term "culture medium" preferably refers to liquid or solid culture media. Thus, for example, the development of a culture medium gradient can be observed from one lateral storage compartment to its opposite lateral storage compartment or from / to the inner storage compartment to one of the lateral storage compartments.

[0118] The sixth aspect of this disclosure relates to another method for culturing cells using a cell culture apparatus, particularly using a cell culture apparatus as described herein, for example, with respect to embodiments of the first aspect. This method can also be performed using a cell culture system as described herein, particularly with respect to the second aspect.

[0119] The method may include the following steps: providing a cell suspension comprising cells and liquid culture medium in at least one or both of two lateral active storage compartments and into the one or more channels. Providing the cell suspension into these channels may be done, for example, by tilting the cell culture apparatus, applying a pressure gradient between the two lateral active storage compartments into which the one or more channels lead, or applying a pressure gradient between at least one of the lateral active storage compartments into which the one or more channels lead and the inner storage compartment into which the channels can lead.

[0120] The method may further include culturing these cells inside the one or more channels. This typically includes growing these cells inside and / or along the channels.

[0121] In some embodiments, the culture medium present in the lateral active storage compartment may be removed first before the cell suspension is provided in at least one or both of the two lateral active storage compartments.

[0122] In some embodiments, the cell suspension may also include a hydrogel precursor in which the cells are suspended, as a liquid culture medium or as a substitute for a liquid culture medium. Preferably, the hydrogel precursor is then solidified after the cell suspension has been provided inside these channels, thereby forming a hydrogel inside these channels, which optionally completely and / or together with the hydrogel present in the hydrogel receiving compartment surrounds the cells. This forms a 3D tissue culture. The hydrogel precursor may be the same as that used in embodiments of the third aspect (e.g., the production of cell culture devices), or it may be different. Thus, in some embodiments, the hydrogel formed inside these channels may be made of the same material as the hydrogel in the hydrogel compartment, or the hydrogel may be made of a different material.

[0123] After the cell suspension has been introduced into these channels, or after or during the culture of these cells within one or more channels, the opposing lateral storage compartments and / or the inner storage compartment may be filled with culture medium. The culture medium may be the same for all opposing lateral storage compartments and the inner compartment, or it may be different. It is also possible that at least one of the lateral storage compartments and the inner storage compartment is not filled with culture medium, i.e., it is empty and may contain only air. As used herein, the term "culture medium" preferably refers to a liquid or solid culture medium. Thus, for example, the development of a culture medium gradient can be observed from one lateral storage compartment to its opposite lateral storage compartment or from / to the inner storage compartment to one of the lateral storage compartments.

[0124] The seventh aspect of this disclosure relates to another method for culturing cells using a cell culture apparatus, particularly using a cell culture apparatus as described herein, for example, with respect to embodiments of the first aspect. This method can also be performed using a cell culture system as described herein, particularly with respect to the second aspect.

[0125] The method may include the step of providing a cell suspension comprising cells and a liquid culture medium in at least one or both of the at least one pair of these opposing lateral storage compartments. These opposing lateral storage compartments may be lateral active storage compartments and / or lateral passive storage compartments, such as the lateral active storage compartments and / or lateral passive storage compartments described herein, for example, with respect to the first aspect of this disclosure.

[0126] The method may further include the step of allowing these cells to settle to the bottom section of the lateral storage compartment. Settling may be performed for a settling time interval. This settling time interval may be, for example, 1 to 15 minutes, particularly 5 to 10 minutes.

[0127] These cells, especially sedimented cells, can be cultured on the bottom section of the lateral storage compartment.

[0128] After the cell suspension has been introduced into at least one or both of the two opposing lateral storage compartments, the remaining opposing lateral storage compartment and / or any of the inner storage compartment may be filled with culture medium. The culture medium may be the same for all opposing lateral storage compartments and the inner compartment, or it may be different. It is also possible that at least one of these lateral storage compartments and the inner storage compartment is not filled with culture medium, i.e., it becomes empty and may contain only air.

[0129] In some embodiments, a majority (i.e., at least 50 vol.%, for example, at least 80 vol.%, for example, at least 90 vol.%, for example, at least 95 vol.%) or all of the liquid culture medium of the cell suspension is removed after the cells have settled from at least one or both of the two opposing lateral storage compartments of the at least one pair. This can be done, for example, by a suitable cannula or syringe.

[0130] After the liquid culture medium has been removed, a different culture medium (e.g., cell culture medium, preferably liquid cell culture medium) may be added to at least one or both of the two opposing lateral storage compartments of the at least one pair, and added to the settled cells.

[0131] Alternatively, after the liquid culture medium has been removed, a hydrogel precursor can be filled into at least one or both of the two opposing lateral storage compartments of the at least one pair covering the settled cells. The hydrogel precursor can then be solidified, thereby forming a hydrogel. The hydrogel precursor can be the same as that used in embodiments of the third aspect (e.g., the production of cell culture apparatus), or it can be different. Thus, in some embodiments, the hydrogel can be made of the same material as the hydrogel in the hydrogel compartment, or it can be made of a different material. Therefore, this hydrogel can (together with the hydrogel in the hydrogel compartment) completely enclose the settled cells. This allows for the creation of 3D tissue cultures.

[0132] In some embodiments, the cell suspension may also include a hydrogel precursor in which the cells are suspended, as a liquid culture medium or as a substitute for a liquid culture medium. Preferably, the hydrogel precursor is then solidified after the cell suspension has been provided to at least one or both of the at least one pair of opposing lateral storage compartments, thereby forming a hydrogel that optionally completely and / or together with the hydrogel present in the hydrogel receiving compartment, encapsulating the cells. This forms a 3D tissue culture. The hydrogel precursor may be the same as that used in embodiments of the third aspect (e.g., the production of cell culture devices), or it may be different. Thus, in some embodiments, the hydrogel formed in at least one or both of the at least one pair of opposing lateral storage compartments may be made of the same material as the hydrogel in the hydrogel compartment, or it may be made of a different material.

[0133] The eighth aspect of this disclosure relates to another method for culturing cells using a cell culture apparatus, particularly using a cell culture apparatus as described herein, for example, with respect to embodiments of the first aspect. This method can also be performed using a cell culture system as described herein, particularly with respect to the second aspect.

[0134] The method may include the following steps: providing a cell suspension comprising a hydrogel precursor and cells in at least one or both of two opposing lateral storage compartments in the at least one pair, particularly in at least one of two lateral active storage compartments and / or two lateral passive storage compartments.

[0135] The method may further include solidifying the hydrogel precursor to form a hydrogel.

[0136] In addition, the method may include culturing these cells inside a solidified hydrogel.

[0137] The hydrogel precursor can be the same as that used in the embodiments of the third aspect (e.g., the production of cell culture devices), or it can be different. Therefore, in some embodiments, the hydrogel can be made of the same material as the hydrogel in the hydrogel compartment, or it can be made of a different material. Thus, the hydrogel can completely enclose the cells. This allows for the creation of 3D tissue cultures.

[0138] In some embodiments, the hydrogel precursor cures before the cells settle into the bottom section of the lateral storage compartments. Alternatively, the hydrogel may cure after the cells have settled into the bottom section of the lateral storage compartments.

[0139] Generally, in all embodiments of the method described herein, particularly with respect to any of the embodiments of aspects five through eight, the cells in the cell suspension may be living cells in some embodiments. These cells may, for example, be derived from human or animal donors. In some embodiments, it is possible that these cells are stem cells.

[0140] In a ninth aspect, this disclosure relates to a cell chip. Such a cell chip may, for example, include a cell culture apparatus as described herein, such as the cell culture apparatus according to the embodiment of the first aspect. Alternatively, the cell chip may include a cell culture system as described herein, such as the cell culture apparatus according to the embodiment of the second aspect.

[0141] Furthermore, the cell chip according to the ninth aspect may include cells cultured in one or more of the following: in the hydrogel, on the surface of the hydrogel, in the cavities of the morphology (specifically the micromorphology) of the hydrogel, in the inner storage compartment, in at least one of the opposing lateral storage compartments (specifically the lateral passive storage compartment and the lateral active storage compartment), and in the one or more channels.

[0142] Therefore, such cell chips can be prepared for use as on-chip laboratory devices in drug assays. Similarly, in some embodiments, the cells in the cell suspension can be live cells. These cells can be derived, for example, from human or animal donors. In some embodiments, these cells may be stem cells.

[0143] In some embodiments, the cell chip can be obtained by any method for culturing cells as described herein (such as the method according to aspects five through eight of this disclosure).

[0144] In a tenth aspect, this disclosure relates to the use of a cell chip, particularly as described in any of the embodiments herein (e.g., embodiments of the ninth aspect). This cell chip can be used to analyze the interaction between cultured cells and a target compound. The target compound can be, for example, a virus, a microorganism such as bacteria or fungi, or a drug such as a protein, peptide, antibody, nucleic acid, small molecule or larger molecule, or antibody-drug conjugate. As used herein, a small molecule is a molecule having a molar mass of less than 1000 Da, and a large molecule is a molecule having a molar mass of 1000 Da or greater. Specific examples of large molecules are, for example, proteins and protein aggregates, enzymes, antigens, or antibodies.

[0145] In some embodiments, the target compound is provided to one or more of the following: provided to the hydrogel, provided to the surface of the hydrogel, provided to the cavities of the micromorphology of the hydrogel, provided to the inner storage compartment, provided to at least one of the opposing lateral storage compartments (particularly lateral passive storage compartments and lateral active storage compartments), and provided to the one or more channels.

[0146] This application can further include monitoring cell states such as viability, cell metabolism, cell migration, cell growth, and permeability (especially permeability across the cell barrier).

[0147] This application could further include monitoring cell signaling, such as paracrine, procrine, or endocrine signaling.

[0148] This use may further include monitoring the location, transport, and / or movement of the compound of interest, such as changes in the location of the target compound from the inner storage compartment to one of these opposing lateral storage compartments.

[0149] In the eleventh aspect, this disclosure further relates to a cell culture apparatus blank. This cell culture apparatus blank can be used for manufacturing cell culture apparatuses, such as the embodiments described with respect to the third aspect. Conversely, the cell culture apparatus blank can be used for manufacturing cell culture apparatuses.

[0150] The cell culture apparatus preform may include a hydrogel compartment configured to be filled with hydrogel. Typically, the hydrogel compartment can therefore be filled with air.

[0151] The cell culture apparatus prototype may further include an inner storage compartment. This inner storage compartment may be defined by an inner storage wall structure and may further include a top opening and an opposing bottom opening. The bottom opening may lead to the hydrogel compartment. The top opening and the bottom opening (and optionally the inner storage compartment as well) may be configured such that the hydrogel compartment is accessible from the top opening, particularly along a straight path from the top opening.

[0152] The cell culture apparatus blank may further include at least a pair of opposing lateral storage compartments that surround the inner storage compartment. In particular, each of these opposing lateral storage compartments leads directly to the hydrogel compartment.

[0153] In some embodiments, these opposing lateral storage compartments may be arranged at the same height as the inner storage compartment. In certain embodiments, at least one or all of these lateral storage compartments may have a height greater than the height of the hydrogel compartment. In some embodiments, at least one or all of these lateral storage compartments may have a height equal to the sum of the height of the hydrogel compartment and a portion or the entire height of the inner storage compartment. It should be understood that the height of the compartment is the vertical extension of the compartment. This vertical direction may extend in the direction of the gravity vector and / or along an axis that extends (particularly vertically) through the center of the bottom opening and / or the center of the top opening of the inner storage compartment. Because the pair of opposing lateral storage compartments surround the inner storage compartment, a relatively large volume of these lateral storage compartments can be achieved. Furthermore, gas exchange, which is crucial for cell culture, can be easily performed.

[0154] Each of these opposing lateral storage compartments has a height of more than 1 mm, particularly more than 2 mm, and even more than 3 mm, which is the extension between the bottom section of the lateral storage compartment and the opening.

[0155] In some embodiments, the height of the hydrogel compartment is at least 500 µm, particularly at least 600 µm, and more particularly at least 700 µm. This height of the hydrogel compartment extends along the vertical direction and can be, for example, the distance between the bottom section and the bottom opening of the inner storage compartment.

[0156] In some embodiments, the volume of the hydrogel compartment can be between 1 and 100 mm. 3 Especially 2 and 10mm 3 between.

[0157] In some embodiments, the hydrogel compartment has an elongated horizontal cross-sectional shape, particularly an elongated circular horizontal cross-sectional shape, such as an ellipse. An elongated shape means that the length-to-width ratio is greater than 1, particularly greater than 1.5. In some embodiments, the hydrogel compartment has an angled horizontal cross-section, particularly having at least three, specifically at least four, more particularly at least five sides and / or angles of a polygonal horizontal cross-section. In some embodiments, the polygonal horizontal cross-section has up to eight, particularly at most six, more particularly at most four sides and / or angles. For example, the hydrogel compartment may have a rectangular horizontal cross-section. The horizontal cross-section is perpendicular to the vertical direction and may extend, for example, parallel to the bottom section and / or the bottom opening of the inner storage compartment.

[0158] In some embodiments, the maximum horizontal width of the hydrogel compartment (i.e., the maximum distance between two opposite peripheral points along the horizontal cross-section) is less than 4 mm, particularly less than 3 mm, and more particularly less than 2.5 mm. In some embodiments, the hydrogel compartment and / or the maximum horizontal width is at least 0.5 mm, particularly at least 1 mm, and more particularly at least 1.5 mm.

[0159] In some embodiments, the opposing lateral storage compartments in each pair are configured such that they are not directly connected to each other. In some embodiments, the opposing lateral storage compartments in each pair are configured such that they are indirectly connected to each other only via the hydrogel compartment. In other words, the opposing lateral storage compartments in each pair are configured such that culture medium exchange between opposing lateral storage compartments (other than bringing fluid into and out of these openings) can only be achieved via the hydrogel (and corresponding hydrogel compartment). In some embodiments, the opposing lateral storage compartments in each pair are configured such that they are indirectly connected to each other only via the hydrogel compartment.

[0160] In some embodiments, the inner storage wall structure includes an inner protrusion. In a particular embodiment, the inner protrusion may define the bottom opening. In some embodiments, the inner protrusion may be disposed between the inner storage compartment and the hydrogel compartment.

[0161] In some embodiments, the inner storage wall structure includes a vertical inner wall section and an inclined inner wall section arranged adjacent thereto. The vertical and inclined inner wall sections may at least partially define at least one of the opposing lateral storage compartments. Alternatively, each lateral storage compartment may be at least partially defined by such vertical and inclined inner wall sections. Such inclined inner wall sections have the advantage that air present in the corresponding lateral storage compartment can be easily vented without being trapped within that compartment. In some embodiments, the inner storage wall section may not contain a vertical inner wall section, but may still include an inclined inner wall section inclined toward the vertical direction of the cell culture apparatus. As noted herein, this vertical direction may be aligned with the gravity vector and / or may be perpendicular to the top opening of the inner storage compartment. For example, the inclined inner wall section can be arranged in cross-section at an angle of 10° to 80°, particularly 25° to 60°, and more particularly 30° to 50° to the vertical direction.

[0162] In some embodiments, the cell culture apparatus blank may also include one or more venting channels. For example, at least one or all of these lateral storage compartments may be connected to such venting channels, with openings that open into the lateral storage compartments. Such venting channels may be arranged such that any remaining gas in a lateral storage compartment is vented through the venting channel during filling (e.g., using liquid culture medium) of the corresponding lateral storage compartment. Thus, the venting channel may be open to the external environment.

[0163] In some general embodiments, these opposing lateral storage compartments can be configured such that any residual gas in the lateral storage compartment is vented out during filling of the corresponding lateral storage compartment (e.g., using a liquid culture medium). This avoids air trapping in the lateral storage compartment.

[0164] In some embodiments, the inclined inner wall section may form an interior angle (i.e., an angle surrounding a portion of the inner storage wall structure), the interior angle being between 90° and 180°, particularly between >90° and <180°, such as 100° to 170°, or 110° to 160°, or 120° to 150°.

[0165] In some embodiments, the inclined inner wall section is configured such that the cross-sectional open area of ​​the lateral storage compartment, at least partially defined by the inclined inner wall section, decreases toward the hydrogel compartment. This means that the cross-section (i.e., the plane parallel to the vertical direction) decreases toward the hydrogel compartment, particularly continuously.

[0166] For example, the inclined inner wall section may have a conical or truncated conical shape, such as a partially conical or partially truncated conical shape. It should be understood that the tip of the cone or truncated cone points towards the hydrogel compartment. This provides a conical or truncated conical path from the lateral storage compartment towards the hydrogel compartment. Therefore, in some embodiments, at least one or all of these opposing lateral storage compartments may include a conical or truncated conical path or a partially conical or partially truncated conical path towards the hydrogel compartment. A partially conical shape is a cross-section of a cone, particularly a cross-section along a plane parallel to or even along the axis of rotation of the cone. The same applies to partially truncated conical shapes.

[0167] In some embodiments, the inclined inner wall section is disposed between the vertical inner wall section and the inner protrusion. The inclined inner wall section may in particular be inclined toward the inner protrusion and the vertical inner wall section. In a particular embodiment, the inner protrusion may be at an angle relative to the vertical inner wall section, for example, an angle of 80° to 100°, particularly 85° to 95°, and more particularly substantially 90°.

[0168] In some embodiments, the inner storage wall structure includes one or more support structures that laterally surround the hydrogel compartment or a portion of the hydrogel compartment. The one or more support structures may be the same as those described in the corresponding embodiments with respect to the first aspect of this disclosure. For example, in some embodiments, the one or more support structures may protrude from the inner protrusion.

[0169] In some embodiments, the inner storage wall structure separates the two opposing lateral storage compartments from the inner storage compartment, specifically such that the two opposing lateral storage compartments and the inner storage compartment are only indirectly fluidly connected via the hydrogel compartment.

[0170] In some embodiments, the hydrogel compartment is arranged between two opposing lateral storage compartments.

[0171] In some embodiments, the hydrogel compartment is surrounded and / or fluidly connected to the inner storage compartment and two opposing lateral storage compartments.

[0172] In some embodiments, the cell culture apparatus preform further includes a hydrogel delivery orifice. The hydrogel delivery orifice may be open toward the hydrogel compartment and may be configured to deliver a hydrogel or hydrogel precursor into the hydrogel compartment. In some embodiments, the hydrogel delivery orifice includes a delivery outlet that leads directly to the hydrogel compartment.

[0173] In some embodiments, the hydrogel delivery orifice has an inlet through which the hydrogel or hydrogel precursor can be filled. The hydrogel delivery orifice can be arranged such that the hydrogel or hydrogel precursor can be delivered directly from the inlet into the hydrogel compartment.

[0174] In some embodiments, the orifice inlet is arranged adjacent to the top opening of the inner storage compartment and / or on the same side of the cell culture apparatus blank as the top opening of the inner storage compartment. In some embodiments, the orifice inlet may be offset from the top opening, particularly laterally offset. In some embodiments, the orifice inlet is defined by the inner storage wall structure.

[0175] In some embodiments, the cell culture apparatus preform further includes a hydrogel buffer compartment. The hydrogel buffer compartment may be configured to receive hydrogel and / or hydrogel precursors during filling. The hydrogel compartment may have access to (particularly direct access to) the hydrogel compartment. For example, the hydrogel buffer compartment may include a hydrogel buffer compartment opening via which the hydrogel buffer compartment is connected to the hydrogel compartment. In a particular embodiment, the hydrogel buffer compartment is separated from the hydrogel delivery orifice. For example, the hydrogel buffer compartment may not be directly connected to the hydrogel delivery orifice, but both the hydrogel delivery orifice and the hydrogel buffer compartment have access to the hydrogel compartment. In some embodiments, the hydrogel buffer compartment may include a vent opening configured to vent air from the hydrogel buffer compartment to the external environment during filling of the hydrogel compartment, and consequently the hydrogel buffer compartment. Typically, the vent opening is different from the hydrogel buffer compartment opening. In some embodiments, the hydrogel buffer compartment may be arranged opposite to the hydrogel delivery orifice, and / or the delivery outlet of the hydrogel delivery orifice may be arranged opposite to the opening of the hydrogel buffer compartment. In some embodiments, the hydrogel buffer compartment may have a volume of 1 to 100 µL, particularly 2 to 20 µL.

[0176] In some specific embodiments, the cell culture apparatus blank includes:

[0177] - Hydrogel compartments, which are configured to be filled with hydrogel;

[0178] - An inner storage compartment defined by an inner storage wall structure, wherein the inner storage compartment includes a top opening and an opposing bottom opening, wherein the bottom opening leads to the hydrogel compartment, and wherein the top opening and the bottom opening are configured such that the hydrogel compartment can be accessed from the top opening; wherein the inner storage wall structure includes an inward protrusion that defines the bottom opening and / or is disposed between the inner storage compartment and the hydrogel compartment;

[0179] - At least one pair of opposing lateral storage compartments, which surround the inner storage compartment and each leads to the hydrogel compartment;

[0180] - A hydrogel delivery orifice, which is open toward the hydrogel compartment and configured to deliver hydrogel or hydrogel precursor into the hydrogel compartment.

[0181] In some embodiments, the cell culture apparatus blank includes an additional pair of opposing lateral storage compartments that surround the inner storage compartment and each opens to the hydrogel compartment, such that the hydrogel compartment is in fluid communication with each of the additional pair of opposing lateral storage compartments. Therefore, in some embodiments, there may also be more than two opposing lateral storage compartments. Typically, two opposing lateral storage compartments form a storage compartment pair. In some embodiments, for example, four lateral storage compartments may be present, i.e., two pairs of opposing lateral storage compartments, each pair consisting of two opposing lateral storage compartments. In some embodiments, it is possible that the cell culture apparatus has a first pair of opposing lateral storage compartments and a second pair of opposing lateral storage compartments. Therefore, in such embodiments, the hydrogel compartment can be accessed from five sides, i.e., from the inner storage compartment and the four lateral storage compartments.

[0182] In a twelfth aspect, this disclosure relates to a cell culture preform system. This cell culture preform system may, for example, include a plurality of cell culture apparatus preforms as described in any of the embodiments herein (such as those in the eleventh aspect). For example, such a cell culture preform system may include at least 2, at least 4, at least 10, at least 20, at least 50, at least 90, or at least 100 cell culture apparatus preforms.

[0183] In some embodiments, the cell culture preform system can be configured as a well plate. Such well plates can include all cell culture apparatus preforms.

[0184] In a thirteenth aspect, this disclosure relates to a method for producing a cell culture apparatus blank, particularly a cell culture apparatus blank as disclosed in any of the embodiments herein. The method may particularly include injection molding.

[0185] In some embodiments, the method may include the following steps:

[0186] -Provide mold components;

[0187] - Inject molten material, particularly molten polymer or molten metal, into the mold assembly;

[0188] - Solidify the injected molten material to form a preform of the cell culture device, and optionally remove the solidified preform of the cell culture device from the mold assembly.

[0189] In some embodiments, the mold assembly includes a bottom portion and a top portion, wherein the top portion includes a mold top protrusion configured to form the inner storage compartment of the cell culture apparatus blank; and wherein the bottom portion includes at least a pair of opposing lateral mold bottom protrusions configured to form the at least one pair of opposing lateral storage compartments of the cell culture apparatus blank.

[0190] It should be understood that in this embodiment or any other embodiment described herein, the mold assembly, and particularly the top and bottom portions of the mold assembly, are configured such that the mold assembly forms a molding cavity having the shape of the cell culture apparatus preform. That is, the mold assembly may have an inverted shape, which is correspondingly a negative of the shape of the cell culture apparatus preform as described herein, particularly with respect to any of the embodiments of the eleventh aspect. The mold assembly may be configured to form the cell culture apparatus preform as described herein, particularly with respect to any of the embodiments of the eleventh aspect. The top and bottom portions may be configured to be movable to separate from each other and may be configured to form the mold assembly together.

[0191] In the fourteenth aspect, this disclosure relates to a mold assembly as described herein, for example, in any of the embodiments relating to the thirteenth aspect.

[0192] In the fifteenth aspect, this disclosure relates to the use of a mold assembly, particularly according to the fourteenth aspect, for manufacturing a cell culture apparatus blank as described herein, particularly with respect to any of the embodiments of the eleventh aspect.

[0193] Non-limiting examples of this disclosure are provided below:

[0194] Example 1. A cell culture apparatus is disclosed, the cell culture apparatus comprising:

[0195] - Hydrogel compartment, which contains hydrogel;

[0196] - Inner storage compartment, the inner storage compartment being defined by an inner storage wall structure (5), the inner storage compartment including a top opening and a bottom opening arranged opposite each other, wherein the bottom opening leads to the hydrogel compartment, and wherein the top opening and the bottom opening are configured such that the hydrogel can be accessed from the top opening;

[0197] - At least one pair of opposing lateral storage compartments, which surround the inner storage compartment and each leads to the hydrogel compartment, such that the hydrogel is in fluid communication with each lateral storage compartment;

[0198] The hydrogel includes a surface that faces and / or is exposed toward the inner storage compartment, particularly through the bottom opening.

[0199] Example 2. A cell culture apparatus according to Example 1, wherein the inner storage wall structure includes an inner protrusion that defines the bottom opening and / or is disposed between the inner storage compartment and the hydrogel compartment.

[0200] Example 3. A cell culture apparatus according to Example 1 or 2, wherein the inner storage wall structure includes a vertical inner wall section and an inclined inner wall section arranged adjacent to the vertical inner wall section, wherein the vertical inner wall section and the inclined inner wall section at least partially define at least one of the opposing lateral storage compartments.

[0201] Example 4. A cell culture apparatus according to Example 3, wherein the inclined inner wall section is configured such that the cross-sectional open area of ​​at least one lateral storage compartment at least partially defined by the inclined inner wall section decreases toward the hydrogel compartment.

[0202] Example 5. A cell culture apparatus according to Example 3 or 4, wherein the inclined inner wall section is arranged between the vertical inner wall section and the inner convex portion.

[0203] Example 6. A cell culture apparatus according to any one of the preceding examples, wherein each lateral storage compartment includes an opening, wherein each lateral storage compartment is configured such that its bottom section is accessible along a straight path from its opening.

[0204] Example 7. A cell culture apparatus according to any one of the foregoing examples, wherein the inner storage wall structure includes one or more support structures that laterally surround the hydrogel or a portion of the hydrogel.

[0205] Example 8. A cell culture apparatus according to any one of Examples 2 to 7, wherein one or more support structures protrude from the inner protrusion.

[0206] Example 9. A cell culture apparatus according to any one of the preceding examples, wherein the inner storage wall structure separates the opposing lateral storage compartments from the inner storage compartment, particularly such that the opposing lateral storage compartments and the inner storage compartment are fluidly connected only indirectly via the hydrogel compartment.

[0207] Example 10. A cell culture apparatus according to any one of the preceding examples, wherein the hydrogel compartments are arranged between these opposing lateral storage compartments.

[0208] Example 11. A cell culture apparatus according to any one of the preceding examples, wherein the hydrogel compartment and / or the hydrogel is surrounded and / or fluidly connected to the inner storage compartment and these opposing lateral storage compartments.

[0209] Example 12. A cell culture apparatus according to any one of the preceding examples, wherein the surface includes a micromorphology having a plurality of cavities, wherein the micromorphology faces and / or is exposed toward the inner storage compartment, particularly via the bottom opening.

[0210] Example 13. A cell culture apparatus according to Example 12, wherein the micromorphology is configured such that each cavity is not undercut.

[0211] Example 14. A cell culture apparatus according to any one of the foregoing examples, the cell culture apparatus further comprising a hydrogel delivery orifice that is open toward the hydrogel compartment and configured to deliver a hydrogel or a hydrogel precursor into the hydrogel compartment.

[0212] Example 15. A cell culture apparatus according to any one of the foregoing examples, wherein the cell culture apparatus further comprises a hydrogel buffer compartment, the hydrogel buffer compartment opening to the hydrogel compartment and being particularly spaced apart from the hydrogel delivery orifice.

[0213] Example 16. A cell culture apparatus according to any one of the preceding examples, wherein the opposing lateral storage compartments of the pair of opposing lateral storage compartments are lateral passive storage compartments, wherein the hydrogel does not have a channel to either of the lateral passive storage compartments, such that compounds can be exchanged between the lateral passive storage compartments only by diffusion through the hydrogel.

[0214] Example 17. A cell culture apparatus according to any one of Examples 1 to 15, wherein the opposing lateral storage compartments of a pair of opposing lateral storage compartments are lateral active storage compartments, wherein the hydrogel defines one or more channels extending between the two lateral active storage compartments and opening in each of the two lateral active storage compartments, wherein the one or more channels are configured for active culture medium exchange between the two lateral active storage compartments.

[0215] Example 18. A cell culture apparatus according to Example 17, wherein the cell culture apparatus additionally includes a pair of opposing lateral passive storage compartments surrounding the inner storage compartment and each opening to the hydrogel compartment, such that the hydrogel is in fluid communication with each lateral passive storage compartment, and wherein the hydrogel does not have a channel to either of the lateral passive storage compartments, such that compounds can be exchanged between the lateral passive storage compartments only by diffusion through the hydrogel.

[0216] Example 19. A cell culture apparatus according to Example 17 or 18, wherein one or more channels have a depth of 10 µm. 2 With 1mm 2 Specifically, 20 µm 2 With 0.8 mm 2 The open area of ​​the cross-section between them.

[0217] Example 20. A cell culture apparatus according to any one of Examples 17 to 19, wherein the one or more channels form a channel network, the channel network particularly including one or more of channel intersections, recesses, protrusions and / or branches.

[0218] Example 21. A cell culture system is provided, the cell culture system comprising a plurality of cell culture devices according to any one of the foregoing examples, wherein preferably, the cell culture system is a well plate.

[0219] Example 22. A method relating to the production of a cell culture apparatus, particularly according to any one of Examples 1 to 20, the method comprising the steps of:

[0220] - Provide a cell culture device blank comprising a hydrogel compartment, an inner storage compartment defined by an inner storage wall structure, and at least a pair of opposing lateral storage compartments surrounding the inner storage compartment and each opening to the hydrogel compartment, wherein the inner storage compartment includes a top opening and opposing bottom openings, wherein the bottom openings open to the hydrogel compartments, and wherein the top openings and the bottom openings are configured such that the hydrogel compartments can be accessed from the top openings;

[0221] - A casting insert, including a casting surface, is introduced through the top opening into the inner storage compartment such that the casting insert covers the bottom opening and the casting surface is exposed to the hydrogel compartment;

[0222] - Fill the hydrogel compartment with the hydrogel precursor such that the hydrogel precursor contacts the casting surface of the casting insert;

[0223] - The hydrogel precursor is cured to form a hydrogel, the hydrogel including a surface facing and / or exposed toward the inner storage compartment, particularly via the bottom opening;

[0224] - The casting insert is removed through the top opening, thereby providing the cell culture apparatus.

[0225] Example 23. The method according to Example 22, wherein the casting surface of the cast insert includes a micromorphology having a plurality of protrusions, and wherein the cast insert is introduced into the inner storage compartment through the top opening such that the cast insert covers the bottom opening and exposes the micromorphology to the hydrogel compartment; and wherein a hydrogel precursor is filled into the hydrogel compartment such that the hydrogel precursor surrounds the protrusions of the micromorphology of the cast insert; and wherein curing the hydrogel precursor forms a hydrogel including a micromorphology having a plurality of cavities.

[0226] Example 24. The method according to Example 22 or 23, wherein the inner storage wall structure of the cell culture apparatus blank includes an inner protrusion that defines the bottom opening and / or is disposed between the inner storage compartment and the hydrogel compartment.

[0227] Example 25. The method according to Example 24, wherein the cast insert is introduced into the inner storage compartment such that the cast insert rests on the inner protrusion of the inner storage wall structure.

[0228] Example 26. The method according to Example 24 or 25, wherein the hydrogel precursor is filled into the hydrogel compartment such that the central portion of the hydrogel precursor is disposed below the casting insert, and the peripheral portion of the hydrogel precursor is disposed below the inner protrusion of the inner storage wall structure.

[0229] Example 27. A method according to any one of Examples 24 to 26, wherein one or more support structures protrude from the inner protrusion, and wherein the one or more support structures retain the hydrogel precursor in the hydrogel compartment during the filling of the hydrogel precursor into the hydrogel compartment.

[0230] Example 28. A method according to any one of Examples 22 to 27, wherein the cell culture apparatus preform includes a hydrogel delivery orifice opening toward the hydrogel compartment, wherein the hydrogel precursor is provided into the hydrogel compartment via the hydrogel delivery orifice during filling.

[0231] Example 29. A method according to any one of Examples 22 to 28, wherein the opposing lateral storage compartments in a pair of opposing lateral storage compartments are laterally active storage compartments, and wherein one or more channels are formed extending through the cured hydrogel and leading to each of the two laterally active storage compartments.

[0232] Example 30. Involves the method according to Example 29, wherein the one or more channels are formed by depleting or degrading portions of the solidified hydrogel, particularly by photodegradation (such as laser ablation or photoinduced depolymerization) or by chemical degradation.

[0233] Example 31. The method according to Examples 22 to 30, wherein the hydrogel precursor comprises cells to be cultured.

[0234] Example 32. A component kit is provided, the component kit comprising:

[0235] - A cell culture apparatus blank, the cell culture apparatus blank comprising a hydrogel compartment, an inner storage compartment defined by an inner storage wall structure, and at least a pair of opposing lateral storage compartments surrounding the inner storage compartment and each opening to the hydrogel compartment, wherein the inner storage compartment includes a top opening and opposing bottom openings, wherein the bottom openings open to the hydrogel compartments, and wherein the top openings and the bottom openings are configured such that the hydrogel compartments can be accessed from the top openings; and

[0236] - A casting insert, wherein the casting insert includes a casting surface, and wherein the casting insert is configured to be introduced into the inner storage compartment through the top opening, such that the casting insert covers the bottom opening and the casting surface is exposed to the hydrogel compartment;

[0237] -Optional hydrogel precursor.

[0238] Example 33. A kit according to Example 32, wherein the cast surface includes a micromorphology having a plurality of protrusions, and wherein the cast insert is configured to be introduced through the top opening into the inner storage compartment such that the cast insert covers the bottom opening and exposes the micromorphology to the hydrogel compartment.

[0239] Example 34. A method for culturing cells using a cell culture apparatus according to any one of Examples 1 to 20 or using a cell culture system according to Example 21, the method comprising the steps of:

[0240] - A cell suspension containing cells and liquid culture medium is provided in the inner storage compartment, particularly via the top opening;

[0241] - Allows the cells to settle onto the surface of the hydrogel, particularly into the plurality of cavities of the micromorphology of the hydrogel, and to culture the cells.

[0242] Example 35. The method according to Example 34, wherein after the cells have settled, a majority of the liquid culture medium is removed from the inner storage compartment, and subsequently the hydrogel precursor is filled into the inner storage compartment covering the settled cells, wherein the hydrogel precursor is then solidified.

[0243] Example 36. A method for culturing cells using a cell culture apparatus according to any one of Examples 1 to 20 or using a cell culture system according to Example 21, the method comprising the steps of:

[0244] - A cell suspension containing cells and liquid culture medium is provided in at least one of the lateral active storage compartments in a pair of opposing lateral storage compartments and provided into the one or more channels, particularly by tilting the cell culture device or by applying a pressure gradient between the lateral active storage compartments to which the one or more channels lead;

[0245] - The cells are cultured inside one or more channels.

[0246] Example 37. A method for culturing cells using a cell culture apparatus according to any of Examples 1 to 20 or a cell culture system according to Example 21, the method comprising the steps of:

[0247] - A cell suspension containing cells and liquid culture medium is provided in at least one of the opposite lateral storage compartments;

[0248] - Allows the cells to settle into the bottom section of the at least one lateral storage compartment;

[0249] - The cells are cultured on the bottom section of the at least one lateral storage compartment.

[0250] Example 38. A method for culturing cells using a cell culture apparatus according to any of Examples 1 to 20 or a cell culture system according to Example 21, the method comprising the steps of:

[0251] - A cell suspension containing hydrogel precursors and cells is provided in at least one of the opposite lateral storage compartments;

[0252] - Solidify the hydrogel precursor to form a hydrogel;

[0253] - The cells are cultured inside a solidified hydrogel.

[0254] Example 39. A method according to Example 38, wherein the hydrogel precursor is cured before or after the cells settle to the bottom section of the at least one lateral storage compartment.

[0255] Example 40. A cell chip comprising a cell culture apparatus according to any one of Examples 1 to 20 or a cell culture system according to Example 21, the cell chip further comprising cells cultured in one or more of the following: in the hydrogel; on the surface of the hydrogel; in the cavity of the micromorphology of the hydrogel; in the inner storage compartment; in at least one of the opposing lateral storage compartments, particularly the lateral passive storage compartment and / or the lateral active storage compartment; and in one or more channels.

[0256] Example 41. Involves a cell chip according to Example 40, which has been obtained by any one of the methods according to Examples 32 to 37.

[0257] Example 42. This relates to the use of a cell chip according to Example 40 or 41 for analyzing the interaction between cultured cells and a target compound, said target compound being, in particular: a virus, a microorganism such as bacteria or fungi, or a drug such as a protein, peptide, antibody, nucleic acid, small molecule, or antibody-drug conjugate.

[0258] Example 43. Relating to the use according to Example 42, wherein the target compound is provided to one or more of the following: provided to the hydrogel, provided to the surface of the hydrogel, provided to the cavity of the micromorphology of the hydrogel, provided to the inner storage compartment, provided to at least one of the opposing lateral storage compartments (particularly lateral passive storage compartments and / or lateral active storage compartments), and provided to the one or more channels.

[0259] Example 44. A cell culture apparatus blank, the blank comprising:

[0260] - Hydrogel compartments, the hydrogel compartments being configured to be filled with hydrogel;

[0261] - An inner storage compartment, defined by an inner storage wall structure, wherein the inner storage compartment includes a top opening and an opposing bottom opening, wherein the bottom opening leads to the hydrogel compartment, and wherein the top opening and the bottom opening are configured such that the hydrogel compartment can be accessed from the top opening;

[0262] - At least one pair of opposing lateral storage compartments, the pair of opposing lateral storage compartments surrounding the inner storage compartment and each opening to the hydrogel compartment.

[0263] Example 45. A cell culture apparatus blank according to Example 44, wherein the inner storage wall structure includes an inner protrusion that defines the bottom opening and / or is disposed between the inner storage compartment and the hydrogel compartment.

[0264] Example 46. A cell culture apparatus blank according to Example 44 or 45, wherein the inner storage wall structure includes a vertical inner wall section and an inclined inner wall section arranged adjacent to the vertical inner wall section, wherein the vertical inner wall section and the inclined inner wall section at least partially define at least one of the opposing lateral storage compartments.

[0265] Example 47. A cell culture apparatus blank according to Example 46, wherein the inclined inner wall section is configured such that the cross-sectional open area of ​​at least one lateral storage compartment at least partially defined by the inclined inner wall section decreases toward the hydrogel compartment.

[0266] Example 48. A cell culture apparatus blank according to Example 46 or 47, wherein the inclined inner wall section is arranged between the vertical inner wall section and the inner protrusion.

[0267] Example 49. A cell culture apparatus blank according to any one of Examples 44 to 48, wherein the inner storage wall structure includes one or more support structures that laterally surround the hydrogel compartment or at least a portion of the hydrogel compartment.

[0268] Example 50. A cell culture apparatus blank according to Examples 45 and 49, wherein one or more support structures protrude from the inner protrusion.

[0269] Example 51. A cell culture apparatus blank according to any one of Examples 44 to 50, wherein the inner storage wall structure separates the opposing lateral storage compartments from the inner storage compartment, particularly such that the opposing lateral storage compartments and the inner storage compartment are fluidly connected only indirectly via the hydrogel compartment.

[0270] Example 52. A cell culture apparatus blank according to any one of Examples 44 to 51, wherein the hydrogel compartment is arranged between two opposing lateral storage compartments.

[0271] Example 53. A cell culture apparatus blank as described in any of Examples 44 to 52, wherein the hydrogel compartment is surrounded and / or fluidly connected to the inner storage compartment and the opposing lateral storage compartment.

[0272] Example 54. A cell culture apparatus blank according to any one of Examples 44 to 53, wherein the cell culture apparatus blank further includes a hydrogel buffer compartment that opens to the hydrogel compartment and is particularly spaced apart from the hydrogel delivery orifice.

[0273] Example 55. A cell culture apparatus blank according to any one of Examples 44 to 54, the cell culture apparatus blank further comprising a hydrogel delivery orifice open toward the hydrogel compartment and configured for delivering a hydrogel or hydrogel precursor into the hydrogel compartment.

[0274] Example 56. A cell culture apparatus blank according to Example 55, wherein the hydrogel delivery orifice has an orifice inlet and is configured such that the hydrogel or hydrogel precursor can be delivered directly from the orifice inlet into the hydrogel compartment.

[0275] Example 57. A cell culture apparatus blank according to Example 55 or 56, wherein the orifice inlet is arranged adjacent to the top opening of the inner storage compartment (4) and / or arranged on the same side of the cell culture apparatus blank as the top opening of the inner storage compartment.

[0276] Example 58. A cell culture apparatus blank according to any one of Examples 44 to 57, wherein the cell culture apparatus blank includes an additional pair of opposing lateral storage compartments surrounding the inner storage compartment and each opening to the hydrogel compartment, such that the hydrogel compartment is in fluid communication with each of the opposing lateral storage compartments in the additional pair of opposing lateral storage compartments.

[0277] Example 59. A cell culture preform system is disclosed, comprising a plurality of cell culture apparatus preforms according to any one of Examples 44 to 58, wherein preferably, the cell culture preform system is a plate with wells.

[0278] Example 60. A method relating to the production of a cell culture apparatus blank according to any one of Examples 44 to 58, the method comprising the following steps

[0279] -Provide mold components;

[0280] - Inject molten material into the mold assembly;

[0281] - Solidify the injected molten material to form the blank of the cell culture device.

[0282] Example 61. A method according to Example 60, wherein the mold assembly includes a bottom portion and a top portion, wherein the top portion includes a mold top protrusion configured to form the inner storage compartment of the cell culture apparatus blank; and wherein the bottom portion includes at least a pair of opposing lateral mold bottom protrusions configured to form the at least one pair of opposing lateral storage compartments of the cell culture apparatus blank.

[0283] Example 62. A method according to Example 61, wherein the bottom portion further includes at least one hydrogel delivery orifice protrusion configured to form the hydrogel delivery orifice of the cell culture apparatus blank.

[0284] Example 63. The method according to Example 61 or 62, wherein the bottom portion further includes one or more recesses configured to form one or more support structures of the cell culture apparatus blank.

[0285] Example 64. A mold assembly is provided that forms a molded cavity having the shape of a cell culture apparatus preform according to any one of Examples 44 to 58. Attached Figure Description

[0286] The invention described herein will be more fully understood from the detailed description and accompanying drawings provided below, which should not be considered as limiting the invention as described in the appended claims. The drawings show:

[0287] Figure 1a is a schematic perspective view of a cell culture apparatus according to an embodiment of the present invention;

[0288] Figure 1b shows a cross-sectional view of the cell culture apparatus shown in Figure 1a along line AA.

[0289] Figure 1c shows a cross-sectional view of the cell culture apparatus of Figure 1a along BB.

[0290] Figure 2a is a perspective view of a cell culture preform system according to an embodiment of the present invention and which can be used in an embodiment of the present invention;

[0291] Figure 2b shows a top view of the cell culture preform system shown in Figure 2a.

[0292] Figure 2c shows a bottom view of the cell culture preform system compared to Figure 2a.

[0293] Figure 3a is a bottom view of a cell culture apparatus blank according to an embodiment of the present invention and which can be used in an embodiment of the present invention;

[0294] Figure 3b shows a top view of the cell culture apparatus blank shown in Figure 3a.

[0295] Figure 3c shows a cross-sectional view of the cell culture apparatus blank along CC in Figure 3b.

[0296] Figure 4a is a top view of a cell culture apparatus blank according to another embodiment of the present invention, which can be used in an embodiment of the present invention;

[0297] Figure 4b shows a cross-sectional view of the cell culture apparatus blank through the lateral storage compartments 8 and 9 of Figure 4a.

[0298] Figure 4c shows a cross-sectional view of the cell culture apparatus blank through the lateral storage compartments 10 and 11 of Figure 4a.

[0299] Figure 4d shows the bottom view of the cell culture apparatus blank as shown in Figure 4a;

[0300] Figure 5 is a top view of a cell culture system according to an embodiment of the present invention;

[0301] Figure 6a is a top view of a cell culture apparatus blank according to another embodiment of the present invention, which may be used in an embodiment of the present invention;

[0302] Figure 6b shows a cross-sectional view of the cell culture apparatus blank along DD.

[0303] Figure 7 is a perspective view of a cell culture apparatus blank according to another embodiment of the present invention, which can be used in embodiments of the present invention;

[0304] Figure 8a is a perspective view of the bottom portion of a mold assembly that can be used to produce a preform of a cell culture device as described herein.

[0305] Figure 8b is a perspective view of the top portion of a mold assembly that can be used to produce a preform of a cell culture device as described herein.

[0306] Figure 9 illustrates the process scheme for preparing a cell culture device and a cell chip according to an embodiment of the present invention;

[0307] Figure 10 is a schematic cross-sectional view of a cell culture apparatus according to another embodiment of the present invention.

[0308] Exemplary embodiments

[0309] Figure 1a Figures 1b to 1c show a cell culture apparatus 1 according to an embodiment of the present disclosure. The cell culture apparatus 1 includes an inner storage compartment 4 disposed centrally, which is laterally defined by an inner storage wall structure 5. As best seen from the cross-sectional views of Figures 1b and 1c, the inner storage compartment 4 includes a top opening 6 and a bottom opening 7 disposed opposite to it. Both the top and bottom openings are defined accordingly by the inner storage wall structure 5. It can be seen that the top opening 6 has a larger open area than the bottom opening 7. The bottom opening 7 leads directly from the inner storage compartment 4 to the hydrogel compartment 2. Both the top opening 6 and the bottom opening 7, as well as the inner storage compartment 4, are configured such that the hydrogel compartment 2 can be accessed from the top opening. This means that, for example, a straight rod such as a cast insert (see Figure 9) can be inserted through the top opening 6 and can access the bottom opening 7 from there.

[0310] Hydrogel compartment 2 is disposed directly below inner storage compartment 4. Furthermore, bottom opening 7 leads directly to the hydrogel compartment. Hydrogel 3 is disposed inside the hydrogel compartment and completely fills it. As can be further seen from Figures 1a and 1b, the hydrogel comprises a micromorphology with multiple cavities such as cavities 12 and 13 (note that only two cavities are referenced, and the micromorphology is not shown in Figure 1a for clarity). The cavities form recesses within the hydrogel 3. It can also be seen that the micromorphology faces and is exposed toward the inner storage compartment 4. Therefore, cells can be introduced into the inner storage compartment 4, for example, as a cell suspension. Allowing cell sedimentation subsequently provides such cells to the cavities of the micromorphology.

[0311] The cell culture apparatus 1 further includes four lateral storage compartments 8, 9, 10, and 11. Lateral storage compartments 8 and 9 form a first pair and are opposite each other, and enclose the inner storage compartment 4, as well as the hydrogel compartment 2 and the hydrogel 3. Figure 1bThe cross-sectional view shown indicates that the hydrogel 3 can be accessed (i.e., contacted) from the two opposing lateral storage compartments 8 and 9. That is, culture medium can be inserted into these opposing lateral storage compartments, and the culture medium subsequently comes into contact with the hydrogel 3. It can be seen that the hydrogel 3 does not have any channels leading to either of the opposing lateral storage compartments 8 and 9. Therefore, these two lateral storage compartments 8 and 9 are considered to be lateral passive storage compartments. Culture medium introduced into either of these two lateral passive storage compartments can only be transported through the hydrogel by diffusion into the hydrogel 3. Therefore, these two opposing lateral passive storage compartments 8 and 9 allow the observation of the diffusion of the target compound through the hydrogel 3, which can mimic the ECM. If, during the assay, the cell barrier is cultured on the surface of the hydrogel exposed to one of the lateral storage compartments 8 or 9, and this exposure leads to barrier disruption and leakage, faster diffusion can be observed, for example, than when the barrier is intact.

[0312] As can be seen from the two cross-sectional views in Figures 1b and 1c, the two opposing lateral storage compartments 8 and 9, together with the hydrogel compartment 2, form a U-shape in cross-section surrounding the inner storage compartment 4. Conversely, the two opposing lateral storage compartments 10 and 11, together with the hydrogel compartment 2, also form a U-shape in cross-section surrounding the inner storage compartment 4. The opposing lateral storage compartments 10 and 11 form a second pair of opposing lateral storage compartments.

[0313] Lateral storage compartments 10 and 11 are also opposite each other and enclose inner storage compartment 4, hydrogel compartment 2, and hydrogel 3. However, in contrast to the lateral passive storage compartments 8 and 9, hydrogel 3 defines channels 18 that all lead to the two lateral storage compartments 10 and 11 and provide direct fluid connection between the two lateral storage compartments. In other words, channels 18 connect the two lateral storage compartments 10 and 11 and pass through hydrogel 3. Therefore, lateral storage compartments 10 and 11 are lateral active storage compartments because they enable active transport of the culture medium through the hydrogel via channels 18.

[0314] Lateral storage compartments 8, 9, 10, and 11 are each partially defined by the outer wall structure 29. Furthermore, each lateral storage compartment 8, 9, 10, and 11 is arranged between the outer wall structure 29 and the inner storage wall structure 5. The cell culture apparatus 1 further includes a bottom section 25 that forms the bottom boundary of the opposing lateral storage compartments and the hydrogel compartment 2. It can be seen that the hydrogel 3 extends between the inner storage wall structure 5 and the bottom section 25. The inner storage wall structure 5 further includes an inward protrusion 14. The inward protrusion 14 is angled relative to the vertical inner wall section of the inner storage wall structure 5 and peripherally surrounds and defines the bottom opening 7. It can be seen that the inward protrusion 14 is configured such that, compared to the top opening 6, the inward protrusion reduces the open area of ​​the bottom opening 7. The inner wall structure 5 further includes support structures 15 and 16 that laterally surround a portion and, in this case, the majority (i.e., greater than 50 vol.%) of the hydrogel 3. Support structures 15 and 16 project from the inner protrusion 14 into the hydrogel compartment 2. Such support structures are advantageous because they help maintain the hydrogel and / or hydrogel precursor within the hydrogel compartment 2 and prevent displacement of the hydrogel and / or hydrogel precursor. Thus, one or more support structures can typically form a shape-locked engagement with the hydrogel. The peripheral segment of the hydrogel 3 is arranged below and in contact with the inner protrusion 14. Furthermore, the central segment of the hydrogel 3, including micromorphology (or, in some embodiments, in other words, a surface with cavities), is arranged below the bottom opening 7.

[0315] Figures 2a and 2c illustrate a cell culture preform system 101 according to an embodiment of the invention and which may be used in some embodiments of the invention. While Figure 2a shows a perspective view, Figure 2b shows a top view through the top openings of the corresponding cell culture device preforms 19, 19', and 19'', and Figure 2c shows a bottom view along the vertical direction. Note that the bottom section has been removed to allow observation of the system interior. The cell culture preform system 101 includes three cell culture device preforms 19, 19', and 19''. Although only cell culture device preform 1' is given reference numerals for clarity, the three cell culture device preforms are identical in their features, differing only in their position within the cell culture preform system 101. Each cell culture device preform includes an inner storage compartment 4 defined by an inner storage wall structure 5. The inner storage wall structure includes an inner protrusion 14 that peripherally defines a bottom opening 7 leading to a hydrogel compartment 2 configured to receive hydrogel. Each cell culture device blank further includes a hydrogel delivery orifice 17 that allows the introduction of hydrogel or a hydrogel precursor into the hydrogel compartment 2. As best seen in Figure 2b, the hydrogel delivery orifice 17 includes an orifice inlet 27 arranged adjacent to the top opening of the inner storage compartment 4. The orifice inlet 27 is further arranged on the same side of the cell culture device blank as the top opening of the inner storage compartment 4. That is, the orifice inlet is not arranged on the same side as the bottom opening 7. In this embodiment, the orifice inlet 27 is further laterally offset from the top opening and is defined and correspondingly constrained by the inner storage wall structure 5. As best seen from the bottom view shown in Figure 2c, the inner storage wall structure 5 further includes six support structures, such as support structures 15 and 16 (only two are referred to for clarity), which protrude from the inner protrusion 14 into the hydrogel compartment 2. The support structures are arranged radially around the bottom opening 7. In this embodiment or any other embodiment as described herein, the support structures may be specifically spaced radially from the bottom opening. The multiple support structures are also spaced apart from each other, such that these support structures define gaps between them. Figure 2c further shows additional details of the hydrogel delivery orifice 17. It can be seen that the hydrogel delivery orifice 17 further includes a delivery outlet 28. The delivery outlet 28 is arranged such that it allows direct delivery of hydrogel or hydrogel precursors into the hydrogel compartment 2. Furthermore, the delivery outlet is arranged such that it leads to the area surrounded by the support structures. As in the embodiment shown in Figure 2c, the cross-section of the hydrogel delivery orifice may be L-shaped.

[0316] Figures 3a and 3c show a cell culture apparatus blank 19 according to an embodiment of the invention and which may be used in some embodiments of the invention. The bottom view shown in Figure 3a shows the bottom opening 7 of the inner storage compartment 4, which leads from the inner storage compartment 4 to the hydrogel compartment 2. The inner storage compartment 4 and the hydrogel compartment 2 are laterally surrounded by two opposing lateral storage compartments 8 and 9, which are fluidly connected to the hydrogel compartment 2 and indirectly fluidly connected to the inner storage compartment 4 via the hydrogel compartment 2. Furthermore, as described with reference to Figures 2a and 2c, the cell culture apparatus black 19 includes an inner storage wall structure 5 having an inner protrusion 14 and support structures 15 and 16 protruding from the inner protrusion 14. Figure 3c shows a cross-sectional view along CC in Figure 3b. It can be seen that the inner storage compartment 4 may be dome-shaped. Furthermore, it can be seen that the inner protrusion 14 narrows the open area of ​​the bottom opening 7 compared to the top opening 6 of the inner storage compartment 2. Two opposing lateral storage compartments 8 and 9 are shown, each arranged between the inner storage wall structure 5 and the outer wall structure 29. The delivery outlet 28 of the hydrogel delivery orifice leads directly to the hydrogel compartment 2, such that the hydrogel or hydrogel precursor is surrounded by a support structure such as the support structure 15 when entering the hydrogel compartment 2.

[0317] Figure 4a shows a top view of a cell culture apparatus blank 19 according to another embodiment of the invention or which may be used in other embodiments of the invention (e.g., for producing a cell culture apparatus according to an embodiment of the invention). Similar to the cell culture apparatus blanks 19 shown in Figures 3a to 3c, the cell culture apparatus blank 19 shown in Figure 4a includes an inner storage compartment 4 defined by an inner storage wall structure 5 and including a top opening 6 and a bottom opening 7 arranged opposite thereto. Furthermore, the cell culture apparatus blank includes a hydrogel compartment through which the bottom opening 7 leads, and the inner storage compartment 4 is laterally surrounded by two opposing lateral storage compartments 8 and 9 of a first pair of opposing lateral storage compartments. However, the cell culture apparatus blank 19 shown here also includes an additional pair of opposing lateral storage compartments 10 and 11. Therefore, the cell culture apparatus blank 19 includes a total of four lateral storage compartments. Opposite lateral storage compartments 8 and 9 form a first pair of opposite lateral storage compartments, and lateral storage compartments 10 and 11 form a second pair of opposite lateral storage compartments. Such cell culture device blanks are particularly advantageous because they allow the production of cell culture devices with a hydrogel accessible from at least five different sites: from the inner storage compartment and from each of the four lateral storage compartments 8, 9, 10, and 11. Furthermore, these cell culture device blanks allow the opposite lateral storage compartments to be used as lateral passive storage compartments while simultaneously using opposite lateral storage compartments 10 and 11 as lateral active storage compartments.

[0318] Figure 4b is a cross-sectional view passing through the center of the inner storage compartment 4 of the cell culture device blank shown in Figure 4a and through a pair of opposing lateral storage compartments 8 and 9. It can be seen that both the two opposing lateral storage compartments 8 and 9, as well as the inner storage compartment 4, are in direct fluid communication with the hydrogel compartment 2. Figure 4c is a cross-sectional view passing through the center of the inner storage compartment 4 of the cell culture device blank shown in Figure 4a and through a pair of opposing lateral storage compartments 10 and 11. It can be seen that these two opposing lateral storage compartments are also in direct fluid communication with the hydrogel compartment 2. Figure 4d shows a bottom view of the cell culture device blank 19. In this embodiment, the inner storage wall structure 5 includes four support structures surrounding at least a portion of the hydrogel compartment 2. Furthermore, the cell culture device blank 19 includes a hydrogel delivery orifice 17. Furthermore, it can be seen that exhaust passages 32 and 33 are connected to lateral active storage compartments 10 and 11 and are configured to exhaust residual air from lateral active storage compartments 10 and 11 to the external environment.

[0319] Figure 5 illustrates a cell culture system comprising a 6x4 matrix of cell culture devices, such as cell culture devices 1, 1', and 1'' (only three are referenced for clarity). The cell culture system is designed as a well plate. In this well plate, rows are designated by reference codes A, B, C, and D, and columns are designated by codes 1, 2, 3, 4, 5, and 6 to distinguish and identify different cell culture devices.

[0320] Figure 6a shows a top view of a cell culture apparatus blank 19 according to another embodiment of the invention or which may be used in other embodiments of the invention (e.g., for producing a cell culture apparatus according to an embodiment of the invention). The cell culture apparatus blank 19 shown in Figure 6a includes an inner storage compartment (see Figure 6b) defined by an inner storage wall structure 5 and including a top opening 6 and a bottom opening 7 arranged opposite thereto. The bottom opening 7 leads to a hydrogel compartment (see Figure 6b). The inner storage compartment 4 is laterally surrounded by two opposing lateral storage compartments 8 and 9 of a first pair of opposing lateral storage compartments. The cell culture apparatus blank 19 includes an additional pair of opposing lateral storage compartments 10 and 11. Therefore, the cell culture apparatus blank 19 includes a total of four lateral storage compartments. Opposite lateral storage compartments 8 and 9 form a first pair of opposite lateral storage compartments, and lateral storage compartments 10 and 11 form a second pair of opposite lateral storage compartments.

[0321] Figure 6b shows a cross-sectional view along DD of the cell culture apparatus blank 19 shown in Figure 6a. It can be seen that the inner storage wall structure 5, which defines and separates the inner storage compartment 4 and the opposing lateral storage compartments 8 and 9, includes a vertical inner wall section 30 and an inclined inner wall section 31. The inner storage wall structure 5 further includes an inner protrusion 14 as described herein. The inclined inner wall section 31 connects to the inner protrusion 14 and the vertical inner wall section 30. Furthermore, the inclined inner wall section is inclined towards both the inner protrusion 14 and the vertical inner wall section 30 at angles less than 180° and greater than 90°. The inner protrusion 14 and the vertical inner wall can be arranged at approximately 90° to each other, as shown in Figure 6b. When liquid culture medium is filled into the lateral storage compartment 9, for example using a pipette, the culture medium level rises slowly, and air trapped adjacent to the hydrogel can be easily expelled through the inclined inner wall section 31. This provides a conical or truncated conical path from the lateral storage compartment toward the hydrogel compartment.

[0322] Figure 7 shows a perspective view of the bottom side of the cell culture apparatus blank 19 with the bottom section removed. The cell culture apparatus blank 19 includes a pair of opposing lateral storage compartments 8 and 9 and a hydrogel compartment 2, which opens to the inner storage compartment via a bottom opening 7. It can be seen that the inner storage wall structure 5 includes an inward protrusion 14, a vertical inner wall section 30, and an inclined inner wall section 31, which allows air removal during the filling of the lateral storage compartments with culture medium. Additionally, the cell culture apparatus blank 19 includes a hydrogel delivery orifice 17, which opens to the hydrogel compartment 2 via its delivery outlet 28. Furthermore, the cell culture apparatus blank 19 shown here includes a hydrogel buffer compartment 34, which also opens to the hydrogel compartment 2. However, as can be seen, the hydrogel buffer compartment is separated from the hydrogel delivery orifice 17 and is therefore a separate element. In this embodiment, the hydrogel buffer compartment 34 is arranged opposite to the hydrogel delivery orifice 17 and its delivery outlet 28. The hydrogel buffer compartment 34 may include a hydrogel buffer compartment opening 35 leading to the hydrogel compartment 2. The hydrogel buffer compartment opening 34 is different from the delivery outlet 28. When the hydrogel or hydrogel precursor is filled into the hydrogel compartment 2 via the hydrogel delivery orifice 17, complete filling of the hydrogel compartment may be difficult. It is possible that the hydrogel compartment is not completely filled and / or the hydrogel or hydrogel precursor has begun to flow out of the hydrogel compartment. By providing such a hydrogel buffer compartment, the filling of the hydrogel compartment can be improved. During filling, excess hydrogel or hydrogel precursor is filled into the hydrogel compartment and can flow into the hydrogel buffer compartment instead of flowing out of the hydrogel compartment 2. The filling of the hydrogel compartment is thus improved and more accurate, and overfilling of the hydrogel compartment can be avoided.

[0323] Figure 8a shows the bottom portion 35 of the mold assembly, which can be formed together with the top portion 36 of the mold assembly shown in Figure 8b. The top and bottom portions have shapes corresponding to a cell culture apparatus preform as described herein. That is, when the top and bottom portions are arranged on top of each other, a mold assembly is formed having the shape of a negative sheet of a cell culture apparatus preform. For example, it can be seen that the bottom portion 35 of the mold assembly includes at least a pair of opposing lateral mold bottom protrusions 37 and 38, which will form at least a pair of opposing lateral storage compartments of the cell culture apparatus preform. Furthermore, the bottom portion 35 of the mold assembly includes at least one hydrogel delivery orifice protrusion 40, which is configured to form a hydrogel delivery orifice of the cell culture apparatus preform when molten material is injected into the mold. The top portion 36 of the mold assembly includes a mold top protrusion 39, which will form an inner storage compartment. It should be understood that in the embodiment described herein or any other embodiment, the mold assembly, and in particular the top and bottom portions of the mold assembly, are configured such that the mold assembly forms a cavity having the shape of a cell culture device preform.

[0324] Figure 9 illustrates a process scheme for an embodiment of a method for producing a cell culture apparatus 1 and a cell chip 26 according to some embodiments of the present disclosure. In a first step a, a cell culture apparatus blank 19 (which may also be part of a cell culture blank system) is provided. The cell culture apparatus blank 19 may be, for example, as shown in Figures 3a to 3c or Figure 4aThe cell culture apparatus blank shown and described in Figure 4d is used. In subsequent step b, a casting insert 20 is provided. The casting insert 20 includes: a base portion having a micromorphology having a plurality of protrusions, such as protrusions 21 and 22 (only two are referred to for clarity); and a further rod protruding from the base portion. The casting insert 20 and the top opening 6 of the cell culture apparatus blank 19 are configured such that the base portion, and correspondingly the casting insert 20, can be inserted into the inner compartment 4. As shown in step b, the casting insert 20 is introduced into the inner compartment 4. The casting insert rests on and contacts the inner protrusion 14 of the cell culture apparatus blank 19, and the micromorphology of the casting insert, and particularly the protrusions, extend through the bottom opening 7 toward the hydrogel compartment, and correspondingly toward the hydrogel compartment. In other words, the micromorphology of the casting insert 20 is exposed to the hydrogel compartment 2. A hydrogel precursor is then inserted into the hydrogel compartment 2, for example, via a hydrogel delivery orifice as described herein. The hydrogel precursor fills the hydrogel compartment 2 in such a manner that it surrounds the protrusions of the micromorphology of the casting insert 20. The hydrogel precursor may also fill any cavity of the casting insert 20, correspondingly filling the micromorphology of the casting insert. Thus, the micromorphology of the casting insert 20 serves as a negative for the micromorphology of the hydrogel. The hydrogel precursor is then cured, for example by heat treatment (such as heating and / or cooling), or by irradiation, for example, with UV light. After the hydrogel precursor has been cured to form the hydrogel 2, the casting insert 20 is removed again via the top opening 6 of the cell culture device 1 thus formed (see step c).

[0325] In the next step or a separate method, the resulting cell culture apparatus 1 (which includes a cell culture apparatus blank 19 and hydrogel 3 housed in a hydrogel compartment 2) can be used to produce cell chip 26. For example, in the illustrated embodiment, a cell suspension including cells 23 in liquid culture medium 24 can be introduced into the inner storage compartment 4. Since the hydrogel 3 and its micromorphology having multiple cavities (such as cavities 12 and 13 corresponding to the protrusions of the casting insert 20) are arranged directly below the bottom opening 7, cells can be selectively supplied to the cavities by micropipette, or the cell suspension can be added to the inner storage compartment and the cells can be allowed to settle. Thus, a cell chip 26 including cell culture apparatus 1 and multiple cells 23 is formed. The cells can then be cultured, for example, to form 3D shapes and structures. For example, cells can form cell barriers on the surface of the hydrogel, correspondingly on the micromorphology. Further, the same or different culture media can be added to the opposing lateral storage compartments 8 and 9, as indicated by the wavy lines.

[0326] Figure 10 illustrates a cell culture apparatus 1 according to another embodiment of the present disclosure. (Refer to Figures 1a to 10) Figure 1c Similar to the cell culture apparatus shown and described, the cell culture apparatus 1 shown here includes an inner storage compartment 4 defined by an inner storage wall structure 5 comprising an inner protrusion 14. The inner storage compartment 4 also includes a top opening 6 and a bottom opening 7, the bottom opening leading to a hydrogel compartment 2 filled with hydrogel 3. Furthermore, the cell culture apparatus 1 includes a pair of opposing lateral storage compartments 8 and 9 surrounding the inner storage compartment 4 and the hydrogel compartment 2. However, in contrast to Figures 1a to 1c, the inner storage wall structure 5 includes an inclined inner wall section 31 immediately adjacent to a vertical inner wall section 30 extending along the vertical direction V, the inclined inner wall section forming an angle α with the vertical inner wall, for example, between 120° and 150°. It can be further seen that both lateral storage compartments 8 and 9 are partially defined by such inclined inner wall sections. In some embodiments, this is effective for each lateral storage compartment. When liquid culture medium is filled into the lateral storage compartment 9, for example, using a pipette, the culture medium level rises slowly, and air trapped by the adjacent hydrogel can be easily expelled through the inclined inner wall section 31. The inclined inner wall section is further angled relative to the inner convex portion 14.

Claims

1. A cell culture apparatus blank (19), the cell culture apparatus blank comprising: - Hydrogel compartment (2), the hydrogel compartment being configured to be filled with hydrogel (3); - An inner storage compartment (4), the inner storage compartment being defined by an inner storage wall structure (5), wherein the inner storage compartment (4) includes a top opening (6) and a bottom opening (7) arranged opposite to each other, wherein the bottom opening (7) leads to the hydrogel compartment (2), and wherein the top opening (6) and the bottom opening (7) are configured such that the hydrogel compartment (2) can be accessed from the top opening (6); - At least one pair of opposing lateral storage compartments (8, 9, 10, 11), the pair of opposing lateral storage compartments surrounding the inner storage compartment (4) and each opening to the hydrogel compartment (2).

2. The cell culture apparatus blank (19) according to claim 1, wherein the inner storage wall structure (5) includes an inner protrusion (14) that defines the bottom opening (7) and / or is disposed between the inner storage compartment (4) and the hydrogel compartment (2).

3. The cell culture apparatus blank (19) according to claim 1 or 2, wherein the inner storage wall structure (5) includes a vertical inner wall section and an inclined inner wall section arranged adjacent to the vertical inner wall section, wherein the vertical inner wall section (30) and the inclined inner wall section (31) at least partially define at least one of the opposing lateral storage compartments (8, 9, 10, 11).

4. The cell culture apparatus blank (19) according to claim 3, wherein the inclined inner wall section (31) is configured such that the cross-sectional open area of ​​at least one lateral storage compartment (8, 9, 10, 11) at least partially defined by the inclined inner wall section (31) decreases toward the hydrogel compartment (2).

5. The cell culture apparatus blank (19) according to claim 3 or 4, wherein the inclined inner wall section (31) is arranged between the vertical inner wall section and the inner protrusion (14).

6. A cell culture apparatus blank (19) according to any one of the preceding claims, wherein the inner storage wall structure (5) includes one or more support structures (15, 16) that laterally surround the hydrogel compartment (2) or at least a portion of the hydrogel compartment (2), wherein preferably the one or more support structures (15, 16) protrude from the inner protrusion (14).

7. A cell culture apparatus blank (19) according to any one of the preceding claims, wherein the inner storage wall structure (5) separates the opposing lateral storage compartments (8, 9, 10, 11) from the inner storage compartment (4), particularly such that the opposing lateral storage compartments and the inner storage compartment are fluidly connected only indirectly via the hydrogel compartment (2).

8. A cell culture apparatus blank (19) according to any one of the preceding claims, wherein the hydrogel compartment (2) is arranged between two opposing lateral storage compartments (8, 9, 10, 11); and / or wherein the hydrogel compartment (2) is surrounded by the inner storage compartment (4) and the opposing lateral storage compartments (8, 9, 10, 11) and / or is fluidly connected to the inner storage compartment and the opposing lateral storage compartments.

9. The cell culture apparatus blank (19) according to any one of the preceding claims, the cell culture apparatus blank (19) further comprising a hydrogel delivery orifice (17) open toward the hydrogel compartment (2) and configured for delivering a hydrogel or hydrogel precursor into the hydrogel compartment (2), wherein preferably the hydrogel delivery orifice (17) has an orifice inlet (27) and is arranged such that the hydrogel or hydrogel precursor can be delivered directly into the hydrogel compartment from the orifice inlet (27).

10. A cell culture apparatus blank (19) according to any one of the preceding claims, wherein the cell culture apparatus blank (19) includes an additional pair of opposing lateral storage compartments (8, 9, 10, 11), the additional pair of opposing lateral storage compartments surrounding the inner storage compartment (4) and each communicating with the hydrogel compartment (2), such that the hydrogel compartment (2) is in fluid communication with each of the opposing lateral storage compartments (8, 9, 10, 11) in the additional pair of opposing lateral storage compartments (8, 9, 10, 11).

11. A cell culture apparatus blank (19) according to any one of the preceding claims, wherein the cell culture apparatus blank further comprises a hydrogel buffer compartment (34) that opens to the hydrogel compartment (2) and is particularly spaced apart from the hydrogel delivery orifice (17).

12. A cell culture apparatus (1), the cell culture apparatus comprising: - Hydrogel compartment (2), the hydrogel compartment containing hydrogel (3); - An inner storage compartment (4), defined by an inner storage wall structure (5), the inner storage compartment (4) including a top opening (6) and a bottom opening (7) oppositely arranged, wherein the bottom opening (7) leads to the hydrogel compartment (2), and wherein the top opening (6) and the bottom opening (7) are configured such that the hydrogel (3) can be accessed from the top opening (6); - At least one pair of opposing lateral storage compartments (8, 9, 10, 11), the opposing lateral storage compartments surrounding the inner storage compartment (4) and each opening to the hydrogel compartment (2), such that the hydrogel (3) is in fluid communication with each lateral storage compartment (8, 9, 10, 11); The hydrogel (3) includes a surface facing the inner storage compartment (4) and / or exposed toward the inner storage compartment, particularly via the bottom opening (7).

13. The cell culture apparatus (1) according to claim 12, wherein the surface comprises a micromorphology having a plurality of cavities (12, 13), wherein the micromorphology faces the inner storage compartment (4) and / or is exposed toward the inner storage compartment, particularly via the bottom opening (7).

14. The cell culture apparatus (1) according to claim 12 or 13, wherein the opposing lateral storage compartments (8, 9) of the pair of opposing lateral storage compartments (8, 9) are lateral passive storage compartments (8, 9), wherein the hydrogel (3) does not have a channel to either of the lateral passive storage compartments (8, 9), such that compounds can be exchanged between the lateral passive storage compartments only by diffusion through the hydrogel.

15. The cell culture apparatus (1) according to claim 12 or 13, wherein the opposing lateral storage compartments (10, 11) of the pair of opposing lateral storage compartments (10, 11) are lateral active storage compartments (10, 11), wherein the hydrogel (3) defines one or more channels (18) extending between the two lateral active storage compartments (10, 11) and opening in each of the two lateral active storage compartments (10, 11), wherein the one or more channels (18) are configured for active culture medium exchange between the two lateral active storage compartments (10, 11).

16. The cell culture apparatus (1) according to claim 15, wherein the cell culture apparatus (1) additionally includes a pair of opposing lateral passive storage compartments (8, 9) surrounding the inner storage compartment (4) and each opening to the hydrogel compartment (2) such that the hydrogel (3) is in fluid communication with each of the lateral passive storage compartments (8, 9), and wherein the hydrogel (3) does not have a channel to any of the lateral passive storage compartments (8, 9) such that compounds can be exchanged between the lateral passive storage compartments (8, 9) only by diffusion through the hydrogel (3).

17. A method for producing a cell culture apparatus (1), particularly a cell culture apparatus (1) according to any one of claims 12 to 16, the method comprising the following steps: - Provides a cell culture device blank (19), particularly a cell culture device blank (1) according to any one of claims 1 to 11, the cell culture device blank comprising a hydrogel compartment (2), an inner storage compartment (4) defined by an inner storage wall structure (5), and at least a pair of opposing lateral storage compartments (8, 9, 10, 11) surrounding the inner storage compartment (4) and each opening to the hydrogel compartment (2), wherein the inner storage compartment (4) comprises a top opening (6) and opposing bottom openings (7), wherein the bottom opening (7) opens to the hydrogel compartment (2), and wherein the top opening (6) and the bottom opening (7) are configured such that the hydrogel compartment (2) is accessible from the top opening (6); - A casting insert (20) including a casting surface is introduced through the top opening (6) into the inner storage compartment (4) such that the casting insert (20) covers the bottom opening (7) and the casting surface is exposed to the hydrogel compartment (2); - Fill the hydrogel precursor into the hydrogel compartment (2) such that the hydrogel precursor contacts the casting surface of the casting insert (20); - The hydrogel precursor is cured to form a hydrogel (3), the hydrogel including a surface facing the inner storage compartment (4) and / or exposed toward the inner storage compartment, particularly via the bottom opening (7); - The casting insert (20) is removed through the top opening (6), thereby providing the cell culture device (1).

18. A method for culturing cells using the cell culture apparatus (1) according to any one of claims 12 to 16, the method comprising the following steps: - A cell suspension containing cells (23) and liquid culture medium (24) is provided in the inner storage compartment (4), particularly via the top opening (6); - Allow the cells (23) to settle onto the surface of the hydrogel (3), particularly into the plurality of cavities (12, 13) of the micromorphology of the hydrogel, and to culture the cells (23).

19. A method for culturing cells using the cell culture apparatus (1) according to any one of claims 15 or 16, the method comprising the steps of: - A cell suspension containing cells and liquid culture medium is provided in at least one of the lateral active storage compartments (10, 11) of the pair of opposing lateral storage compartments (10, 11) and to the one or more channels (18), particularly by tilting the cell culture device (1) or by applying a pressure gradient between the lateral active storage compartments (10, 11) to which the one or more channels (18) lead; - The cells are cultured inside one or more channels (18).

20. A method for culturing cells using the cell culture apparatus (1) according to any one of claims 12 to 16, the method comprising the steps of: - A cell suspension containing cells and liquid culture medium is provided in at least one of the opposite lateral storage compartments (8, 9, 10, 11); - Allow the cells to settle to the bottom section (25) of the at least one lateral storage compartment (8, 9, 10, 11); - The cells are cultured on the bottom section (25) of the at least one lateral storage compartment (8, 9, 10, 11).

21. A method for culturing cells using the cell culture apparatus (1) according to any one of claims 12 to 16, the method comprising the steps of: - A cell suspension containing hydrogel precursors and cells is provided in at least one of the opposite lateral storage compartments (8, 9, 10, 11); -The hydrogel precursor is solidified to form a hydrogel (3); - The cells are cultured inside the solidified hydrogel (3).

22. A cell chip (26) for the purpose of analyzing the interaction between cultured cells (23) and a target compound, said target compound being, in particular, a virus, a microorganism such as bacteria or fungi, or a drug such as a protein, peptide, antibody, nucleic acid, small molecule or antibody-drug conjugate, said cell chip (26) comprising a cell culture apparatus (1) according to any one of claims 12 to 16, said cell chip (26) further comprising cells (23) cultured in one or more of the following: in said hydrogel (3); on said surface of said hydrogel; in said cavity (12, 13) of said micromorphology of said hydrogel (3); in said inner storage compartment (4); in said opposing lateral storage compartments (8, 9, 10, 11), particularly said lateral passive storage compartments (8, 9) and / or said lateral active storage compartments (10, 11); and in said one or more channels (18).

23. A method for producing a cell culture apparatus blank (19) according to any one of claims 1 to 11, the method comprising the following steps -Provide mold components; - Inject molten material into the mold assembly; - Solidify the injected molten material to form the blank of the cell culture device.

24. The method of claim 23, wherein the mold assembly includes a bottom portion (36) and a top portion (37), wherein the top portion (37) includes a mold top protrusion (40) configured to form the inner storage compartment of the cell culture device blank; and wherein the bottom portion includes at least a pair of opposing lateral mold bottom protrusions (38, 39) configured to form the at least one pair of opposing lateral storage compartments of the cell culture device blank.

25. A mold assembly that forms a molded cavity having the shape of a cell culture apparatus preform according to any one of claims 1 to 11.

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