Device for creating three-dimensional cell structures in a hydrogel, microsystem and / or microfluidic cell culture platform, and process for producing the device

Abstract

The invention relates to a device (10) for creating three-dimensional cell structures in a hydrogel, comprising at least one gel chamber (12) having an access opening (14) for filling of the gel chamber (12) with hydrogel, characterized in that the device (10) further comprises at least one first fluid chamber (16) for filling with fluid including at least one biological cell, at least one first channel (18) which connects the first fluid chamber (16) and the gel chamber (12) in a fluid-communicating manner, and at least one second channel (20) which is connected to the gel chamber (12) in a fluid-communicating manner, wherein the gel chamber (12) connects the first channel (18) to the second channel (20) in a fluid-communicating manner. The device (10) can be adapted to the respective requirements flexibly and with a low degree of complexity.

Description

[0001] Device for generating three-dimensional cell structures in a hydrogel, microsystem and / or microfluidic cell culture platform and method for producing the device

[0002] The invention relates to a device for producing three-dimensional cell structures in a hydrogel, a microsystem and / or microfluidic cell culture platform and a method for producing the device.

[0003] Recent technological advances and legislation are enabling regulatory authorities and pharmaceutical companies to use more and better in vitro models for preclinical studies during drug development. The aim is to ethically reduce the use of animal models and achieve lower drug failure rates during clinical trials, e.g. due to human-specific side effects that cannot be monitored using classic mouse models. The models are usually created on the basis of advanced biomimetic and physiologically relevant three-dimensional (3D) cell aggregates, including spheroids, organoids, assembloids and stem embryos, which are collectively referred to as three-dimensional cell structures below.Their use in robust high-throughput research, drug development, and clinical diagnostics remains very limited due to their high heterogeneity, low physiological complexity, lack of standardization, and insufficient accessibility to systematic and automated screening methods, e.g., multiomics and image analysis. To understand these biological

[0004] To solve these problems, three-dimensional cell structures have been integrated into microsystems and microfluidic cell culture platforms, which can also be called chips. The combination is then referred to as a microphysiological system or organ-on-chip.

[0005] However, the shape and structure of existing microphysiological systems or organ-on-chips are unsuitable for screening applications.

[0006] From Claire A . Dessalles et al 2022 Biofabrication 14 015003 , DOI 10 . 1088 / 1758 - 5090 / ac2baa , a microfluidic device is known which has a hydrogel block with a channel . One end of the channel opens into a first reservoir and the opposite end of the channel opens into a second reservoir . Fluid can be pumped from the first reservoir into the second reservoir, whereby a syringe acts as a pump or different fill levels in the reservoirs are proposed to generate the pressure. The channel is created by means of a needle which is pierced from the outside through one of the reservoirs into the other reservoir. The hydrogel is then deployed around the needle so that the channel forms in the hydrogel when the needle is withdrawn.

[0007] The object of the invention is to provide a device for producing three-dimensional cell structures in a hydrogel, which can be adapted flexibly and with little effort to the respective

[0008] requirements can be adapted.

[0009] The problem is solved by the features of the independent claims. Advantageous further developments are the subject of the dependent claims and the following description.

[0010] In a device for producing three-dimensional cell structures in a hydrogel, comprising at least one gel chamber with an access opening for filling the gel chamber with hydrogel, it is provided according to the invention that the device further comprises at least one first fluid chamber for filling with fluid which has at least one biological cell, at least one first channel which connects the first fluid chamber and the gel chamber in a fluid-communicating manner, and at least one second channel which is fluid-communicatingly connected to the gel chamber, wherein the gel chamber connects the first channel to the second channel in a fluid-communicating manner.

[0011] This provides a device which has a gel chamber from which at least two channels branch off. A first channel provides a fluid-communicating connection with a first fluid chamber. The second channel is fluid-connected to the first channel through the gel chamber. This means that in order for fluid to flow between the first channel and the second channel, it must flow through the gel chamber. The alignment of the channels can be chosen arbitrarily, so that, for example, the first channel and the second channel can enclose any angle to one another. In contrast to the prior art, it is not necessary for the first channel and the second channel to be aligned in a line with one another. Furthermore, neither the first channel nor the second channel has an extension along the connection direction between the gel chamber and the first fluid chamber or a further fluid chamber which opens into the external environment.This means that no needle has to be used to produce the channel, which is pierced from the outside, so that production can be carried out with comparatively little effort. This also increases the scalability of the device and the device can be flexibly adapted to the respective requirements with little effort. Furthermore, it is not necessary for the device to have an outer wall through which a needle would have to be pierced, but further devices can be arranged around the device. According to some embodiments, it is conceivable that the second channel can connect the gel chamber to the first fluid chamber in a fluid-communicating manner or that the second channel can connect the gel chamber to a second fluid chamber of the device in a fluid-communicating manner.According to some embodiments, a second fluid chamber can be provided which can be in fluid communication with the first fluid chamber. The fluid can then flow from the first fluid chamber through the first channel, the gel chamber, the second channel to the second fluid chamber and vice versa. The flow of the fluid can be carried out by tilting or inclining the device so that the level of the fluid in one of the two fluid chambers is higher than in the other fluid chamber. This can also happen reciprocally so that the fluid can flow back and forth alternately between the first fluid chamber and the second fluid chamber.

[0012] According to other embodiments, the second channel may extend from the gel chamber to the first fluid chamber. The device may then be tilted such that the fluid flows from the first fluid chamber through the first channel to the gel chamber into the second channel. The tilt angle of the device may then be changed such that the fluid flows back into the first fluid chamber through the second channel.

[0013] In this way, the fluid can flow by rocking or pivoting the device. This can be accomplished using simple means.

[0014] According to some embodiments, it is conceivable that the first channel can open into the gel chamber between the access opening and at least one first bottom section of the gel chamber opposite the access opening.

[0015] The opening of the first channel does not necessarily have to be directly adjacent to the first floor section of the gel chamber. Instead, the opening of the first channel can be at a distance from the first

[0016] bottom section. This allows hydrogel to be introduced into the gel chamber below the opening of the first channel.

[0017] For example, if a wire-like object protrudes from the first channel into the gel chamber, an opening can be created in the hydrogel when the hydrogel is introduced into the gel chamber.

[0018] According to some embodiments, it is conceivable that the second channel can open into the gel chamber between the access opening and at least one second bottom section of the gel chamber opposite the access opening.

[0019] The opening of the second channel does not necessarily have to open into the gel chamber directly above the second floor section of the gel chamber. Instead, the opening of the second channel can be spaced apart from the second floor section. In this way, hydrogel can be introduced into the gel chamber below the opening of the second channel. For example, if a wire-like object protrudes from the second channel into the gel chamber, an opening can be created in the hydrogel when the hydrogel is introduced into the gel chamber.

[0020] If the first channel is also at a distance from a first base section assigned to it, a wire-like body can extend through the gel chamber from the first channel to the second channel. When hydrogel is introduced into the gel chamber, a through-opening can be created in the hydrogel in this way if the wire-like body extends from the second channel through the gel chamber into the first channel or vice versa. According to some embodiments, it is conceivable that the first channel and / or the second channel can be designed as a microchannel, wherein the microchannel can have a diameter in the range between 0.1 pm and 1000 pm, preferably between 1 pm and 1000 pm, more preferably between 20 pm and 500 pm, most preferably between 50 pm and 250 pm.

[0021] The diameters of the channels can thus be freely selected within a wide range. This increases design freedom, allowing the device to be used flexibly in various embodiments for a variety of applications.

[0022] Due to the diameters in the micrometer range, the flow velocity of the fluid is correspondingly low, so that biological cells from the fluid can arrange themselves on the surfaces of the hydrogel.

[0023] According to some embodiments, it is conceivable that the gel chamber may have a cylindrical shape.

[0024] The cylinder can be designed such that the bottom surface of the gel chamber is circular, and the outer surface of the cylinder represents the side walls of the gel chamber into which the channels open. The access opening can also be circular and located opposite the bottom surface.

[0025] However, this does not exclude the possibility that the gel chamber may have a cuboid shape in some other embodiments.

[0026] According to some embodiments, it is conceivable that the device may further comprise a wire-like element which extends through the gel chamber and may be removable from the gel chamber and is preferably designed to be deformable.

[0027] When introducing hydrogel into the gel chamber, the hydrogel cannot flow into the volume of the wire-like element. After the hydrogel has dried, the wire-like element can be removed from the hydrogel and from the gel chamber, creating a through-hole in the hydrogel. The through-hole can then have the shape of the section of the wire-like element that was enclosed by the hydrogel.

[0028] According to some embodiments, it is conceivable that the wire-like element can have a smaller diameter than the first channel and / or the second channel and can be arranged to be movable in the first channel and / or the second channel.

[0029] This allows the wire-like element to extend through the gel chamber to the first channel and / or the second channel. The through-opening created in the hydrogel after removal of the wire-like element is then fluidly connected to the first channel and / or the second channel.

[0030] According to some embodiments, it is conceivable that the wire-like element can extend between the first channel and the second channel.

[0031] The fluid can thus flow from the first fluid chamber through the first channel into the passage opening of the hydrogel. In this way, biological cells, for example, can be introduced into the passage opening of the hydrogel to accumulate there. The cells can form a particularly three-dimensional cell structure there.

[0032] According to some embodiments, it is conceivable that the wire-like element can have a surface with a polyamide material.

[0033] The use of the polyamide material can reduce friction between the hydrogel and the wire-like element. This can facilitate the removal of the wire-like element from the hydrogel and reduce the likelihood of damage to the hydrogel's through-hole.

[0034] According to some embodiments, it is conceivable that the gel chamber can be filled with a hydrogel, wherein the hydrogel can have at least one through-opening which can be fluidly connected to the first channel and / or the second channel.

[0035] As explained above, the through-hole can be created by inserting a wire-like element into the gel chamber before the hydrogel is inserted into the gel chamber. The through-hole can be created by removing the wire-like element. By providing a hydrogel with the through-hole, a three-dimensional cell structure can be created in the through-hole using the fluid, which can be used for other purposes.

[0036] According to some embodiments, it is conceivable that the gel chamber may have at least one insert piece, which may include a support region for supporting the hydrogel, which may extend into the hydrogel, and a holding region for holding the insert piece, which preferably extends outside the hydrogel.

[0037] The holding area can thus be arranged partly inside and partly outside the hydrogel. The insert can thus support the hydrogel, while the hydrogel can be removed from the gel chamber for further investigations after the three-dimensional cell structures have been created in the through-hole. The insert can be grasped by the holding area for this purpose.

[0038] According to some embodiments, it is conceivable that the hydrogel can have at least one biological cell at least at an edge region of the through-opening. The through-opening is understood in this disclosure to be channel-like. The walls of the channel, which are arranged within the hydrogel, can be supplied with the at least one biological cell by wetting with the fluid from the first fluid chamber. The through-opening preferably has a circular cross-section, so that the biological cells can accumulate along a cylindrical surface within the hydrogel and form a correspondingly shaped three-dimensional cell structure.

[0039] According to some embodiments, it is conceivable that at least the first fluid chamber can contain a fluid that can have at least one biological cell.

[0040] The fluid can thus be used to provide a transport medium for the at least one biological cell in order to transport the at least one biological cell, for example, into the passage opening.

[0041] According to some embodiments, it is conceivable that the first fluid chamber can have at least one inlet and / or outlet opening for filling and / or draining fluid.

[0042] In some embodiments, the inlet and / or outlet opening can be arranged opposite a bottom surface of the fluid chamber. The fluid can then be introduced into the first fluid chamber or removed from the first fluid chamber through the inlet and / or outlet opening, for example, using a pipette.

[0043] According to some embodiments, it is conceivable that the device can have at least one base body and a base body, wherein the base body can extend between the access opening and the base body. The base body can, for example, be placed on the base body and connected to it in a material-locking and / or form-fitting manner. In some other embodiments, the base body and the base body can be formed as a single piece.

[0044] According to some embodiments, it is conceivable that the base body can have at least one chamber wall section of the gel chamber and / or the fluid chamber.

[0045] The chamber wall section can, for example, be formed between the gel chamber and / or the fluid chamber. The base body can then provide at least some of the walls of the device's chambers. This can simplify the manufacture of the device if the base body is attached to the base body.

[0046] According to some embodiments, it is conceivable that the device can comprise at least one layer which can comprise at least one hole, the edge of which can delimit the first channel and / or the second channel and / or a portion of the gel chamber and / or a portion of the fluid chamber, wherein the at least one layer can preferably comprise a film material.

[0047] The device can therefore have at least one layer, so that the production of the first channel, the second channel, the section of the gel chamber and / or a section of the fluid chamber can be simplified. For example, a film can be provided which has corresponding holes. By using one film, a large number of holes can be provided in the film, so that the film can be used for the production of a large number of devices at the same time. The costs of producing the device can thus be reduced. Furthermore, by using the at least one layer, the number of devices can be scalable as required.

[0048] According to some embodiments, it is conceivable that the base body can have a base plate, wherein the at least one layer can be arranged between the base plate and the base body.

[0049] The base plate can be designed as a base plate for the base body. At least one layer described above can be arranged between the base plate and the base body. This means that the base plate can delimit the device at the bottom.

[0050] According to some embodiments, it is conceivable that the base plate can be transparent at least in some areas.

[0051] This allows, for example, a measurement of the hydrogel to be performed by irradiating it with light transmitted through the base plate. In this case, the light source can be located beneath the base plate, for example.

[0052] According to some embodiments, it is conceivable that the base plate can comprise a glass material and / or a plastic material, preferably an object carrier for a microscope and / or a microscope cover glass.

[0053] The base body can thus be arranged, for example, on a microscope slide or a microscope cover glass. Microscopy of the hydrogel in the at least one gel chamber of the

[0054] The device can thus be simplified.

[0055] According to some embodiments, it is conceivable that at least one first layer can be arranged between the base body and the first channel and / or the second channel and can have at least one first hole, the edge of which can delimit a first portion of the gel chamber, and at least one second hole, the edge of which can delimit a first portion of the fluid chamber.

[0056] The first layer can, for example, delimit the first channel and / or the second channel transversely to the layer plane, away from the first hole and / or the second hole. This means that a portion of the first layer can then form a channel wall for the first channel and / or the second channel. This can further simplify the manufacture of the device. Furthermore, costs can also be reduced.

[0057] According to some embodiments, it is conceivable that the first layer can be arranged between the base body and at least one second layer and the second layer can have at least one third hole, the edge of which delimits at least the first channel, the second channel and a second section of the gel chamber.

[0058] The second layer can, for example, provide wall sections of the first channel, the second channel, and the second section of the gel chamber with the edge of the third hole parallel to the layer plane. This can further simplify the manufacture of the device.

[0059] According to some embodiments, it is conceivable that the second layer can preferably be arranged between the first layer and at least one third layer, and the third layer has at least one fourth hole, which can have at least a third portion of the gel chamber.

[0060] The third layer can, for example, be directly connected to the

[0061] base body and provide a section of the gel chamber near the bottom, which can be referred to as the third section. The third section can provide a distance between the base body and the mouth of the channels, so that a hydrogel which is introduced into the gel chamber can be arranged between the mouths of the channels and the base body. This can ensure that when a wire-like body is arranged in the gel chamber and the channels, the through-opening to be produced is closed transversely to the direction of extension of the wire-like element. The layers explained above can, in some embodiments, be combined in one piece in various combinations. Furthermore, the layers can also be combined in one piece with the base body or the base body. For example, the first layer can be combined in one piece with the base body and / or the third layer can be combined in one piece with the base body.For the manufacture of the device, the second layer could then be provided between the base body and the main body. Alternatively or additionally, the second layer can be integral with the first layer and / or the main body. Further alternatively or additionally, the second layer can be integral with the third layer and / or the base body.

[0062] The invention further relates to a microsystem or a microfluidic cell culture platform, which is designed in particular as a microtiter plate or slide, comprising a large number of devices according to the preceding description, wherein the access openings of the devices can be arranged next to one another in a matrix-like manner. By providing the microsystem or the microfluidic cell culture platform, a large number of the devices explained above can be provided cost-effectively. Due to the large number of devices, a large number of different combinations of hydrogels and three-dimensional cell cultures can be used. Furthermore, a large number of different fluids with different biological cells can also be used simultaneously on one platform.

[0063] According to some embodiments, it is conceivable that the microsystem or the microfluidic cell culture platform can have a number of devices in the range of 2 to 1152, preferably 2, 4, 8, 16, 32, 60, 128, 512 or 1152, most preferably 128 or 512.

[0064] The number of devices can thus be varied within a wide range. It cannot be ruled out that the number of devices could even be greater than 1152. If two fluid chambers are connected to the gel chamber via the two channels, the number of openings can be three times greater than the number of devices. For example, the microsystem can contain 6 to 3456 openings for chambers.

[0065] This can increase compatibility with current state-of-the-art systems.

[0066] Further advantages and effects, as well as further developments of the microsystem and / or the microfluidic cell culture platform, arise from the advantages and effects, as well as further developments of the device described above. To avoid repetition, reference is made to the preceding description in this regard.

[0067] The invention further relates to a method for producing a device according to the preceding description and / or a microsystem and / or a microfluidic cell culture platform according to the preceding description, comprising at least the following steps: providing a chamber part which comprises at least one gel chamber wall of a gel chamber with an access opening for filling the gel chamber with hydrogel, at least one fluid chamber wall of a first fluid chamber for filling with fluid which has at least one biological cell, at least one first channel wall of a first channel which connects the first fluid chamber and the gel chamber in a fluid-communicating manner, and at least one second channel wall of a second channel which is fluid-communicatingly connected to the gel chamber, wherein the gel chamber fluid-communicatingly connects the first channel to the second channel, and a base part,which has at least one third channel wall of the first channel and at least one fourth channel wall of the second channel; connecting the chamber part to the base part such that the first channel wall is arranged opposite the third channel wall and the second channel wall is arranged opposite the fourth channel wall.

[0068] The method thus describes an exemplary embodiment for producing a device as explained above. The base body explained above can, for example, be part of the chamber part or the entire chamber part. Analogously, the base body explained above can be part of the base part or the entire base part. The first channel wall and / or the second channel wall can extend in a first plane which can be aligned substantially parallel to the access opening. Accordingly, the third channel wall and / or the fourth channel wall are then also aligned substantially parallel to the access opening. However, the angle of the first channel wall and / or the second channel wall can be selected as desired. For example, a first channel wall and / or second channel wall can also be provided which can be aligned perpendicular to a plane of the access opening.The first channel wall and the second channel wall can have different angles to each other.

[0069] The connection between the chamber part and the base part can be materially bonded and / or form-fitting. According to some embodiments, it is conceivable that the method can further comprise at least the following step: inserting at least one wire-like body between the chamber part and the base part such that the wire-like body extends at least between the first chamber wall and the third chamber wall and the second chamber wall and the fourth chamber wall.

[0070] The wire-like body can therefore already be used during production of the device explained above, if, for example, the first channel and / or the second channel are not yet fully formed. For example, the first channel or the second channel can initially be open transversely to the corresponding direction of extension between the fluid chamber and the gel chamber. Due to the open design of the first channel to or the second channel, the wire-like body can be inserted into the corresponding channel. As the method continues, the first channel or the second channel can then be closed by connecting the chamber part to the base part. The wire-like body can then only be accessible through the gel chamber and / or the first and / or second fluid chamber.

[0071] Accordingly, if, for example, a hydrogel is introduced into the gel chamber, the wire-like body can be removed through the first and / or second fluid chamber.

[0072] According to some embodiments, it is conceivable that the method can further comprise at least the following step: fastening at least one first layer, which has at least one first hole, the edge of which delimits a first section of the gel chamber, and at least one second hole, the edge of which delimits a first section of the fluid chamber, to a base body comprising at least one chamber wall of the gel chamber such that the chamber wall continues at least part of the edge of the first hole, in order to provide the chamber part. By providing and fastening the first layer to the base body, a side wall of at least the first channel can be provided at the same time, which side wall can in particular be oriented substantially parallel to the access opening of the base body. With regard to the advantages and effects, reference is made below to the description of the device explained above.

[0073] According to some embodiments, it is conceivable that the method may further comprise at least the following step: attaching at least one third layer to the base body, which layer has at least one fourth hole, which has at least a third section of the gel chamber.

[0074] By providing and attaching the third layer to the base body, a further side wall of at least the first channel can be provided simultaneously, which can in particular be oriented substantially parallel to the access opening of the base body. Further, with regard to the advantages and effects, reference is made to the above-explained description of the device.

[0075] According to some embodiments, it is conceivable that the method may further comprise at least the following step: attaching at least one second layer, which has at least one third hole, the edge of which delimits at least the first channel, the second channel and a second section of the gel chamber, to the first layer in such a way that the first section of the gel chamber is connected to the second section of the gel chamber in a fluid-communicating manner and / or to the third layer in such a way that the second section of the

[0076] The gel chamber is fluidly connected to the third section of the gel chamber. The second layer can thus be attached either to the first layer or to the third layer or to the first layer of the third layer simultaneously. By forming the layers, the number of devices can be scaled as desired. This is especially true when the layers are made of foils.

[0077] According to some embodiments, it is conceivable that the chamber part and the base part can be connected by means of the second layer.

[0078] The chamber part and the base part can then be manufactured separately and connected to each other via the second layer.

[0079] According to some embodiments, it is conceivable that the method further comprises at least the following step: producing the chamber part and / or the base part by means of an additive manufacturing process, preferably a 3D printing process, a subtractive manufacturing process, in particular milling, laser and / or water jet cutting, or a casting process, preferably an injection molding process.

[0080] Using the manufacturing processes mentioned, the chamber part and / or the base part can be manufactured efficiently and cost-effectively.

[0081] According to some embodiments, it is conceivable that the method further comprises at least the following step: producing the first hole, the second hole, the third hole and / or the fourth hole by a cutting method, preferably laser cutting, by an etching method and / or by a punching method. The holes can thus be produced quickly, with high precision and cost-effectively. Furthermore, a plurality of holes can be provided in a short time using the methods mentioned.

[0082] According to some embodiments, it is conceivable that the second layer is attached to the third layer and / or the first layer is attached to the second layer by lamination. The lamination provides a material-to-material attachment between the second layer and the third layer or between the second layer and the first layer. The lamination can be carried out manually or mechanically. In the latter case, large areas can be laminated at once, so that the manufacturing costs for a large number of devices can be concentrated simultaneously. The individual device can thus be provided cost-effectively.

[0083] According to some embodiments, it is conceivable that the method further comprises at least the following step: fixing the third layer to a base plate after fixing the second layer to the third layer.

[0084] The base part can thus be constructed in a modular manner. The individual layers can first be provided with the corresponding holes and then fastened to one another. This simplifies the production process for the channels and the insertion of the wire-like body. According to some embodiments, it is conceivable that the method, after the base body has been fastened to the first layer, further comprises at least the following steps: inserting an insert piece which has a support region for supporting a hydrogel, which can extend into the hydrogel, and a holding region for holding the insert piece, which can extend outside the hydrogel, into the gel chamber in such a way that the support region extends at least into the second section of the gel chamber.

[0085] According to some embodiments, the support region can be formed from at least one strut that extends away from the holding region. The strut can have a free end portion that can be oriented away from the holding region. At least one hook element that can extend away from the end portion can be arranged on the end portion. In some embodiments, the at least one hook element can extend in a direction that is transverse to an extension direction of the strut between the holding region and the end portion.

[0086] The advantages and effects of the insert have already been explained above with regard to the device. To avoid repetition, reference is made to the above description.

[0087] According to some embodiments, it is conceivable that the method, after fastening the base body to the first layer, further comprises at least the following steps: filling at least one gel chamber with a hydrogel such that the hydrogel encloses the wire-like body at least within the gel chamber; and removing the wire-like body from the gel chamber and the first channel and / or the second channel. By means of these method steps, a through-opening can be produced in the hydrogel which was defined by the wire-like body. Further advantages and effects have already been explained above with regard to the device. To avoid repetition, reference is therefore made to the above description.According to some embodiments, it is conceivable that the method further comprises at least the following steps: filling at least one first fluid chamber with a fluid comprising at least one biological cell; tilting the device and / or the microtiter plate such that the fluid flows from the first fluid chamber through the first channel to the hydrogel.

[0088] In this way, by tilting the device, a flow of fluid from the first fluid chamber to the gel chamber can be effected. The fluid then flows through the first channel into the through-opening of the hydrogel. The biological cells that may be contained in the fluid can then attach themselves to the walls of the through-opening of the hydrogel and form three-dimensional cell structures. Further advantages and effects relating to these features have already been explained for the device mentioned above, so that in order to avoid repetition, reference is made to the description above. Further advantages and effects, as well as further developments of the method as a whole, arise from the advantages and effects, as well as further developments of the device described above, the microsystem and / or the microfluidic cell culture platform described above.To avoid repetition, reference is made to the previous description in this regard.

[0089] The invention is described below using an exemplary embodiment with the aid of the accompanying drawings. They show:

[0090] Figures 1a and 1b show a schematic representation of the device according to a first exemplary embodiment; Figure 2 shows a schematic representation of the device according to a second exemplary embodiment;

[0091] Figure 3a-d schematic representations of the layers;

[0092] Figure 4a-c shows a schematic representation of the device according to a third embodiment;

[0093] Figure 5a-c schematic representations of the insert;

[0094] Figure 6 is a schematic representation of a micro system;

[0095] Figure 7a-f shows a schematic representation of a

[0096] Embodiment of the manufacture of the device according to the second embodiment;

[0097] Figure 8 is a flow chart of the process for producing the

[0098] device; and

[0099] Figure 9a-d shows a schematic representation of the use of the

[0100] device .

[0101] The device for producing three-dimensional cell structures in a hydrogel is designated in its entirety by the reference numeral 10, as shown in Figure 1a.

[0102] According to Figure 1a, the device 10 has at least one gel chamber 12 having an access opening 14. Figure 1a shows a schematic plan view of the gel chamber 12 or the device 10.

[0103] Furthermore, the device 10 has at least one first fluid chamber (number 16) into which a fluid can be introduced. The fluid can contain at least one biological cell. Preferably, the fluid contains a plurality of biological cells. Furthermore, the fluid can be a nutrient solution for biological cells.

[0104] The first fluid chamber 16 is fluidly connected to the gel chamber 12 via a first channel 18. Fluid disposed in the first fluid chamber 16 can flow into the gel chamber 12 through the first channel 18. The device 10 has at least one second channel 20 fluidly connected to the gel chamber 12.

[0105] The first channel 18 and the second channel 20 are fluidly connected to one another via the gel chamber 12. This means that the second channel 20 and the first channel 18 are separated or spaced apart from one another by the gel chamber 12.

[0106] In some embodiments, the second channel 20 may be directly connected to the first fluid chamber 16 in a fluid-communicating manner. This means that the second channel 20 may extend from the gel chamber 12 to the first fluid chamber 16 and open into the first fluid chamber 16.

[0107] In some other embodiments, the device 10 may include a second fluid chamber 22 into which the second channel 20 may open. Therefore, in this embodiment, the second channel 20 may fluidly connect the gel chamber 12 to the second fluid chamber 22.

[0108] Figure 1b shows the latter embodiment in a cross-sectional view. The first channel 18 and / or second channel 20 are arranged along a straight line in this embodiment. In some other embodiments, the first channel 18 and the second channel 20 can also be arranged at an angle to each other that does not correspond to 180°. Furthermore, the first channel 18 and / or second channel 20 can also have a non-linear extension.

[0109] According to Figure 1b, the device 10 can have a base body 28 and a base body 30. The base body 18 can have the gel chamber 12 with the access opening 14. The chambers extend from an upper side of the base body 28 to a lower side of the base body 28. The base body 30 can have the first channel 18 and the second channel 20. Furthermore, the base body 30 can have sections of the gel chamber 12 and the first fluid chamber 16 and / or the second fluid chamber 22.

[0110] The first channel 18 can open into the gel chamber 12 at a position spaced from a first bottom portion 24 of the gel chamber 12. In this embodiment, the opening of the first channel 18 into the gel chamber 12 is arranged higher than the first bottom portion 24.

[0111] The second channel 20 can open into the gel chamber 12 at a position spaced from a second bottom portion 26 of the gel chamber 12. In this embodiment, the opening of the second channel 20 into the gel chamber 12 is arranged higher than the second bottom portion 26.

[0112] In this exemplary embodiment, the base body 30 can be formed in one piece and completely encompass the first channel 18 and the second channel 20. The base body 30 can be manufactured, for example, by a subtractive manufacturing process, such as milling, an additive manufacturing process, such as 3D printing, or a casting process. Figure 2 shows a further exemplary embodiment of the device 10.

[0113] In this exemplary embodiment, the base body 30 does not directly comprise the first and second channels 18, 20. Instead, at least one layer 32, 34, 36 is arranged between the base body 28 and the base body 30.

[0114] A first layer 32 may be attached to the base body 28. A third layer 36 may be attached to the base body 30. The first layer 32 and the third layer 36 may be connected to each other via a second layer 34. The second layer 34 may be the first

[0115] Attach layer 32 to the third layer 36 or vice versa.

[0116] The first channel 18 or second channel 20 can extend between the first layer 32 and the third layer 36. In this exemplary embodiment, a respective section of the first layer 32 can provide an upper channel wall for each of the first channel 18 and the second channel 20, in particular a first channel wall and a second channel wall. Furthermore, a respective section of the third layer 36 can provide a respective lower channel wall for each of the first channel 18 and the second channel 20, in particular a third channel wall and a fourth channel wall.

[0117] The base body 28 can form a chamber part 52 of the device with at least one layer 32, 34, 36. Furthermore, the base body 30 can form a base part 54 with at least one other of the layers 32, 34, 36. In particular, the second layer 34 can be part of either the chamber part 52 or the base part 54.

[0118] In Figures 3a to 3b, the first layer 32, the second layer 34 and the third layer 36 are shown schematically.

[0119] According to Figure 3a, the first layer 32 can have at least one first hole 38, the edge of which can delimit a first portion of the gel chamber 12. Furthermore, the first layer 32 can have at least one second hole 40, 42, the edge of which can delimit a first portion of the fluid chamber. In this exemplary embodiment, the edge of the hole 40 can delimit a first portion of the first fluid chamber 16. Furthermore, in this exemplary embodiment, an edge of the hole 42 can delimit a first portion of the second fluid chamber 22.

[0120] The first layer 32 can be aligned with the base body 28 such that the first hole 38 is fluidly connected to the sections of the gel chamber 12 formed in the base body 28.

[0121] Analogously, the second hole 40 in this embodiment can be arranged to be fluidly communicating with the sections of the first fluid chamber 16.

[0122] The second hole 40, 42 may have a smaller diameter than the portions of the first fluid chamber 16 and the second fluid chamber 22, respectively, which are arranged in the base body 28.

[0123] The diameter of the first hole 38 may correspond to the diameter of the sections of the gel chamber 12 arranged in the base body 28. The edge of the first hole 38 may thus be aligned with a chamber wall section of the gel chamber 12 provided by the base body 28.

[0124] Figure 3b shows a representation of the second layer 34. The second layer 34 can have a third hole 44, the edge of which delimits a second section of the gel chamber 12, the first channel 18, the second channel 20, and at least the first fluid chamber 16. Furthermore, it can be provided that the edge of the third hole 44 can also delimit a second section of the second fluid chamber 22. The second layer 34 can be arranged such that the second section of the gel chamber 12 is fluidly connected to the first section of the gel chamber 12, i.e., the first hole 38.

[0125] Preferably, the second portion of the gel chamber 12 is aligned with the first portion of the gel chamber 12.

[0126] Analogously, the second portion of the first fluid chamber 16 can also be arranged in fluid communication with the first portion of the fluid chamber 16. At least a portion of the edge of the hole 44 that defines the second portion of the first fluid chamber 16 can be aligned with a portion of the edge of the second hole 40 that defines the first portion of the fluid chamber 16.

[0127] Figure 3c shows a representation of the third layer 36. The third layer 36 can have a fourth hole 46, the edge of which can define a third section of the gel chamber 12. Furthermore, the third layer can have a fifth hole 48, 50, which can define a third section of the first fluid chamber 16 or a third section of the second fluid chamber 22, respectively.

[0128] The third layer 36 can be arranged such that the fourth hole 46 is arranged in fluid communication with the second section of the gel chamber 12. Similarly, the fifth hole 48, 50 can be arranged such that the third sections of the first fluid chamber 16 and the second fluid chamber 22 are arranged in fluid communication with the respective second sections.

[0129] Figure 3b shows a schematic plan view in which the second layer 34 and the third layer 36 are connected to each other.

[0130] The third layer 36 can provide channel walls for the first channel 18 and the second channel 20. A wire-like body 56 can be arranged on these channel walls, which extends at least in

[0131] TI extends into the gel chamber 12. In this embodiment, the wire-like body 56 can extend from the second portion of the first fluid chamber 16 through the first channel 18 into the second portion of the gel chamber 12. From there, the wire-like body 56 can extend further into the second channel 22 and the second portion of the second fluid chamber 22.

[0132] The wire-like body 56 may be made of a polyamide material. Alternatively, the wire-like body 56 may have a coating of a polyamide material, so that a surface may be made of a polyamide material.

[0133] Figures 4a and 4b show a schematic cross-sectional view of the device 10 with the wire-like body 56.

[0134] According to Figure 4a, the access opening 14 can be oriented upwards, and the wire-like body 56 can be arranged below the access opening 14 in the gel chamber 12. A hydrogel 58 can be filled into the gel chamber 12. The hydrogel 58 can be introduced into the gel chamber 12 through the access opening 14 and can be distributed above and below the wire-like body 56 in the gel chamber 12.

[0135] Furthermore, the hydrogel 58 can also extend laterally of the wire-like body 58 and enclose the wire-like body 58 within the gel chamber 12 from all sides.

[0136] The wire-like body 56 can be removed from the device 10 via the first fluid chamber 16 or the second fluid chamber 22. In doing so, the wire-like body 56 can be pulled out along the first channel 18 or the second channel 20. The wire-like body 56 can be flexible so that it can be bent in order to be able to change direction between the direction of extension of the first channel 18 or the second channel 20 and the direction of extension of the first fluid chamber 16 or the second fluid chamber 22.

[0137] If the wire-like body 56 has a surface that has only a low coefficient of friction with the hydrogel 58, for example a polyamide surface, damage to the interfaces between the hydrogel 58 and the wire-like body 56 can be avoided.

[0138] After removing the wire-like body 56, a through-opening 59 is arranged in the hydrogel. The through-opening 59 can provide a fluid-communicating connection between the first channel 18 and the second channel 20.

[0139] This is schematically illustrated in the cross-sectional view of Figure 4c. Furthermore, an embodiment is shown here in which an insert 60 is inserted before the hydrogel 58 is introduced into the gel chamber 12. The insert 60 can be positioned above the first channel 18 or the second channel

[0140] 20 extend to the base sections 24, 26. The insert piece 60 is shown in more detail in Figures 5a to 5c.

[0141] According to Figure 5a, the individual piece 60 can have a holding area 62 that can be adapted to a shape of the gel chamber 12. In the embodiment according to Figure 5a, the holding area 62 has an angular shape, in particular a square shape.

[0142] A support region 64 may extend away from the holding region 62. The support region 64 may have support elements 65, which may be fastened at one end to the holding region 62. The opposite end of the support elements 65 may be formed from a free end portion. Figure 5b shows a further embodiment of the insert piece 60, in which the holding region 62 is round, in particular circular.

[0143] Figure 5c shows the insert 60 in the state schematically illustrated in cross-section in Figure 4c. At least the support region 64 is enclosed by hydrogel 58. The support elements 65 support the hydrogel 58. The holding region 62 can be arranged entirely outside the hydrogel 58 or at least partially within the hydrogel 58.

[0144] The support elements 65 or the support region 64 can be designed such that the through-opening 59 can extend between the support elements 65. This prevents the support region 64 or the support elements 65 from blocking the through-opening 59.

[0145] With the insert 60, the hydrogel 58 can be removed from the gel chamber 12 and handled for further examinations.

[0146] Figure 6 shows a microsystem 66 comprising a plurality of devices 10. The devices 10 are arranged in a matrix-like manner in the microsystem 66. In this exemplary embodiment of the microsystem 66, the devices 10 each comprise a wire-like body 56 that extends through the respective gel chamber 12. A hydrogel can thus be filled into the respective gel chambers 12.

[0147] Furthermore, a fluid containing at least one biological cell can be introduced into the fluid chambers 16, 22 in order to introduce biological cells into the through-opening 59 of the hydrogel 58 after the wire-like body 56 has been removed from the hydrogel 58. Instead of the microsystem 66, a microfluidic cell culture platform, in particular a microtiter plate or a slide with a plurality of devices 10, can also be used.

[0148] Figures 7a to 7f show an exemplary embodiment of the production of a device 10 or a microsystem 66. In this exemplary embodiment, according to Figure 7a, the second layer 34 can first be provided as a film into which the third hole 44 has been punched or otherwise inserted. The third layer 36 can then be provided, into which the fourth hole 46 and the fifth holes 48, 50 are introduced.

[0149] The second layer 34 can be bonded to the third layer 36, for example, by means of a material bond. This can be achieved, for example, by lamination.

[0150] After the second layer 34 has been attached to the third layer 36, as shown in Figure 7b, the third layer 36 can be connected to a base plate, which can form the base body 30. In this exemplary embodiment, the second layer 34, the third layer 36, and the base body 30 can form the base part 54.

[0151] The attachment of the third layer 36 to the base body 30 can be provided by a material connection, for example an adhesive connection.

[0152] According to Figure 7c, a wire-like body 56 can be inserted into the third hole 44 of the second layer 34. The wire-like body 56 can extend through the second section of the gel chamber 12. Furthermore, the wire-like body 56 can be arranged in the first channel 18 and / or the second channel 20.

[0153] According to Figure 7d, the first layer 32 can then be applied to the second

[0154] Layer 34 can be arranged and secured. The attachment can also be achieved by lamination. The first layer 32 and the third layer 36 can provide opposing channel walls of the first channel 18 and / or the second channel 20.

[0155] According to Figure 7e, a base body 28 can be attached to the first layer 32. The attachment can also be provided by a material-to-material connection, for example by an adhesive connection.

[0156] Figure 7 f shows the manufactured device 10 or the manufactured microsystem 66. The first fluid chambers 16, gel chambers 12, and second fluid chambers 22 provided by the base body 28 are fluidly connected to the respective first sections, second sections, and third sections provided by the first layer 32, the second layer 34, and the third layer 36.

[0157] The thickness of the layers 32, 34, 36 can be in the micrometer range. Accordingly, the first channel 18 and / or the second channel 20 can have a diameter in the range of 0.1 pm and 1000 pm, preferably between 1 pm and 1000 pm, more preferably between 20 pm and 500 pm, most preferably between 50 pm and 250 pm.

[0158] Figures 7a to 7f show the production of a single device 10. However, the components shown in Figures 7a to 7f can have a plurality of gel chambers 12, first fluid chambers 16 and second fluid chambers 22 as well as first channels 18 and second channels 20. The area of ​​the layers can therefore be chosen arbitrarily and the number of holes 38-50 can be adapted to the corresponding number of devices 10 to be produced. The number of chambers in the base body 28 can also be adapted analogously. Furthermore, the area of ​​the base body 30 can also be adapted to the size and number of devices 10.

[0159] Figure 8 shows a flowchart illustrating various embodiments of the method for manufacturing a device 10 or a microsystem 66 and / or a microfluidic cell culture platform according to the above description. According to a step 102, the chamber portion 52 and the base portion 54 are provided.

[0160] The chamber part 52 has at least one gel chamber wall of the gel chamber 12 and the access opening 14. Furthermore, the chamber part 52 has a fluid chamber wall of the first fluid chamber 16.

[0161] With respect to the first channel 18 and the second channel 20, the chamber portion 52 has at least a first channel wall and a second channel wall. The first channel wall may define one side of the first channel 18. The second channel wall may define one side of the second channel 18.

[0162] The bottom part 54 can have a third channel wall of the first channel 18 and a fourth channel wall of the second channel 20.

[0163] In some embodiments, the chamber part 52 can be formed in one piece. For this purpose, the chamber part 52 can be manufactured, for example, according to the optional sub-step 114 of step 102, using an additive manufacturing process. This can be, for example, an additive manufacturing process, a subtractive manufacturing process, or a casting process.

[0164] An additive manufacturing process can be, for example, a 3D printing process. A subtractive manufacturing process can be, for example, a milling process, in particular using a CNC milling machine, or a cutting process using laser and / or

[0165] A water jet could be used. A casting process could, for example, be an injection molding process.

[0166] According to a further embodiment, the chamber part 52 and the base part 54 can be formed in several parts.

[0167] The layers 32, 34, 36 are formed as foils. The foils can then be attached to the base body 28 or the base body 30.

[0168] According to an optional step 116, the first hole 38, the second hole 40, 42, the third hole 44, the fourth hole 46, and / or the fifth hole 48 can be produced in the layers 32, 34, 36 by a cutting process, an etching process, and / or a punching process. An example of a cutting process is preferably laser cutting. Step 116 can be performed simultaneously with sub-step 114 and / or step 102.

[0169] According to the optional step 112, the second layer 34 can be attached to the first layer 32 and / or to the third layer 36.

[0170] The second layer 34 can then be attached to the first layer 32 such that the second section of the gel chamber 12 is fluidly connected to the first section of the gel chamber 12. In this case, an edge of the hole 44 that delimits the second section can expediently be aligned with a section of the edge of the hole 38 that delimits the first section. The second layer 34 can then alternatively or additionally be attached to the third layer 36 such that the second section of the gel chamber 12, which is provided by the second layer 34, is fluidly connected to the third section of the gel chamber 12, which is provided by the third layer 36. Ideally, an edge of the third hole 44, which is arranged on the second section, is aligned with a section of the edge of the hole 46 that delimits the third section.In some other embodiments, the chamber part 52 can comprise the base body 28 and the first layer 32. For this purpose, according to the optional step 108, the at least one first layer 32 can be attached to the base body 28. The first layer 32 can be attached such that an edge of the first hole 38 of the first layer 32 and the at least one chamber wall of the gel chamber 12 are aligned. The chamber wall of the gel chamber 12 thus continues the edge of the first hole 38.

[0171] According to a further optional step 110, the at least one third layer 36 can be attached to the base body 30.

[0172] The first layer 32 or the third layer 36 can be attached by lamination.

[0173] Furthermore, the base part 54 can optionally have the base body 30 and the third layer 36.

[0174] The chamber part 52 and / or the base part 54 can comprise the second layer 32. The second layer 32 can also be divided equally between the chamber part 52 and the base part 54.

[0175] It is conceivable that the second layer 32 can be divided into at least two layers, for example, parallel to the layer plane, wherein at least one sub-layer can be arranged on the chamber part 52 and the other layer on the base part 54. The two sub-layers can both have the same surface area as the second layer 32.

[0176] Alternatively or additionally, the second layer 32 can also be divided into at least two sections perpendicular to the layer plane. The sections can each have a smaller surface area than the second layer 32. A first section can be arranged on the chamber part 52, and a second section can be arranged on the base part 54.

[0177] A mixed form of the two embodiments of the second layer 32 explained above is also conceivable.

[0178] The step 110 may further comprise the optional sub-step 118, in which the third layer is attached to a base plate. The base plate may be part of the base body 30. The step 118 may be carried out after the second layer has been attached to the third layer. Alternatively, the third layer may also be attached without prior attachment of the second layer to the third

[0179] layer to be attached to the base plate.

[0180] Furthermore, according to an optional step 106, it can be provided to arrange at least one wire-like body between the chamber part 52 and the base part 54. The wire-like body 56 can, for example, be inserted between the first channel wall or third channel wall and / or between the second channel wall and the fourth channel wall. For this purpose, the wire-like body 56 can, for example, be placed on the third channel wall and / or on the fourth channel wall. In some other exemplary embodiments, the wire-like body 56 can, for this purpose, be placed on the first channel wall and / or on the second channel wall. In step 104, the chamber part 52 can be connected to the base part 54. Further, the first channel wall is arranged opposite the third channel wall. Further, the second channel wall is arranged opposite the fourth channel wall. The first channel wall and the second channel wall then delimit the first channel 18 on two opposite sides.The third channel wall and the fourth channel wall delimit the second channel 18 on two opposite sides.

[0181] The fastening of the chamber part 52 to the base part 54 can be carried out by means of the second layer 34, as already explained above.

[0182] According to the optional step 120, inserts can be inserted into the gel chamber 12. The insert is inserted into the gel chamber 12 such that the support region of the insert is arranged closer to the base part than the holding region. In a further step 122, hydrogel can be filled into the gel chamber 12. The hydrogel can enclose a wire-like body 56 arranged in the gel chamber 12, at least within the gel chamber 12.

[0183] The insert can be inserted before or after the insertion of a hydrogel into the gel chamber 12. At least a portion of the holding region of the insert can be arranged outside the filled hydrogel. According to a further optional step 124, the wire-like body 56 can be removed from the gel chamber 12. Furthermore, the wire-like body 56 can be removed from the first channel 18 or the second channel 20. In this way, the through-opening 59 can be produced in the hydrogel 58.

[0184] According to optional step 126, the first fluid chamber 16 can be filled with a fluid containing at least one biological cell. Conveniently, the fluid contains a plurality of biological cells.

[0185] According to the optional step 128, the fluid can flow from the first fluid chamber 16 through the first channel 18 to the hydrogel by tilting the device 10 and / or the microtiter plate. This is shown in more detail in Figures 9a to 9d.

[0186] In Figure 9a, the fluid 68 has been filled into the first fluid chamber 16. Due to the small diameter of the first channel 18, the fluid 68 flows very slowly through the first channel 18. As a result, the level of the fluid 68 in the first fluid chamber 16 is higher than in the second fluid chamber 22. The fluid 68 therefore flows to the second fluid chamber 22 by gravity.

[0187] This effect can be enhanced by tilting the device 10 or the microsystem 66. The flow continues until the fluid levels in the first fluid chamber 16 and the second fluid chamber 22 have reached the same level.

[0188] This is shown as an example in Figure 9b.

[0189] The inclination of the device 10 can then be changed so that the flow of the fluid 68 is reversed. This is illustrated in Figures 9c and 9d.

[0190] Alternatively, the second channel 20 can be provided to open into the first fluid chamber 16. In this case, a second fluid chamber 22 can be omitted. The inclination of the device 10 or the microsystem 66 can then be performed in such a way that the flow of the fluid 68 through the second channel back into the first fluid chamber 16 is promoted.

[0191] The example described above does not limit the invention in any way. Rather, the invention can be modified in many ways. All of the features of the invention described above can be essential to the invention alone or in combination with one another.

[0192] The present invention may also be characterized by the following embodiments:

[0193] 1. Device (10) for producing three-dimensional cell structures in a hydrogel, comprising at least one gel chamber (12) with an access opening (14) for filling the gel chamber (12) with hydrogel, characterized in that the device (10) further comprises at least one first fluid chamber (16) for filling with fluid which has at least one biological cell, at least one first channel (18) which connects the first fluid chamber (16) and the gel chamber (12) in a fluid-communicating manner, and at least one second channel (20) which is fluid-communicatingly connected to the gel chamber (12), wherein the gel chamber (12) fluid-communicatingly connects the first channel (18) to the second channel (20).

[0194] 2. Device (10) according to embodiment 1, characterized in that the second channel (20) fluidly connects the gel chamber (12) to the first fluid chamber (16) or that the second channel (20) fluidly connects the gel chamber (12) to a second fluid chamber (22) of the device (10).

[0195] 3. Device (10) according to one of the preceding embodiments, characterized in that the first channel (18) opens into the gel chamber (12) between the access opening (14) and at least one first bottom section (24) of the gel chamber (12) opposite the access opening (14) and / or the second channel (20) opens into the gel chamber (12) between the access opening (14) and at least one second bottom section (26) of the gel chamber (12) opposite the access opening (14). Device (10) according to one of the preceding embodiments, characterized in that the first channel (18) and / or the second channel (20) is designed as a microchannel, wherein the microchannel has a diameter in the range between 0.1 pm and 1000 pm, preferably between 1 pm and 1000 pm, more preferably between 20 pm and 500 pm, most preferably between 50 pm and 250 pm, and / or that the gel chamber (12) has a cylindrical shape.Device (10) according to one of the preceding embodiments, characterized in that the device (10) further comprises a wire-like element which extends through the gel chamber (12) and out of the gel chamber.

[0196] (12) is removable and is preferably designed to be deformable, wherein the wire-like element preferably has a smaller diameter than the first channel (18) and / or second channel (20) and is movably arranged in the first channel (18) and / or the second channel (20), and / or the wire-like element further preferably extends between the first channel (18) and the second channel (20) and / or the wire-like element further preferably has a surface with a polyamide material.Device (10) according to one of the preceding embodiments, characterized in that the gel chamber (12) is filled with a hydrogel, wherein the hydrogel has at least one through-opening which is fluidly connected to the first channel (18) and / or the second channel (20) and / or the gel chamber (12) preferably has at least one insert piece which comprises a support region for supporting the hydrogel, which extends into the hydrogel, and a holding region for holding the insert piece, which preferably extends outside the hydrogel, and / or the hydrogel further preferably has at least one biological cell at least at an edge region of the through-opening.Device (10) according to one of the preceding embodiments, characterized in that at least the first fluid chamber (16) contains a fluid which has at least one biological cell, wherein the first fluid chamber (16) preferably has at least one inlet and / or outlet opening for filling and / or draining fluid. Device (10) according to one of the preceding embodiments, characterized in that the device (10) has at least one base body (28) and a bottom body.

[0197] (30), wherein the base body (28) extends between the access opening (14) and the base body (30), wherein the base body (28) preferably has at least one chamber wall section of the gel chamber (12) and / or the fluid chamber and / or the device (10) preferably has at least one layer which has at least one hole, the edge of which delimits the first channel (18) and / or the second channel (20) and / or a section of the gel chamber (12) and / or a section of the fluid chamber, wherein the at least one layer preferably comprises a film material, wherein the

[0198] The base body (30) further preferably has a base plate, wherein the at least one layer is arranged between the base plate and the base body (28) and / or the base plate is further preferably transparent at least in some areas and / or the base plate comprises a glass material and / or a plastic material, preferably a microscope slide and / or a microscope cover glass, and / or at least one first layer (32) is arranged between the base body (28) and the first channel (18) and / or the second channel (20) and has at least one first hole (38), the edge of which delimits a first section of the gel chamber (12), and at least one second hole (40, 42), the edge of which delimits a first section of the fluid chamber, wherein the first layer (32) is preferably arranged between the base body (28) and at least one second layer (34) and the second layer (34) has at least one third hole (44),the edge of which delimits at least the first channel (18), the second channel (20) and a second portion of the gel chamber (12), wherein further the second layer (34) is preferably arranged between the first (32) layer and at least one third layer (36), and the third layer (36) has at least one fourth hole (46) which has at least a third portion of the gel chamber (12). Microsystem (66) and / or microfluidic cell culture platform, in particular microtiter plate or slide, comprising a plurality of devices (10) according to one of the preceding embodiments, wherein the access openings (14) of the devices (10) are arranged next to one another in a matrix-like manner, wherein the microsystem and / or microfluidic cell culture platform preferably has a number of devices (10) in the range from 2 to 1152, preferably 2, 4, 8, 16, 32, 60, 128, 512 or 1152, most preferably 128 or 512,A method (100) for producing a device (10) according to one of embodiments 1 to 8 and / or a microsystem and / or microfluidic cell culture platform according to the preceding embodiment, comprising at least the following steps:

[0199] Providing (102) a chamber part (52) which has at least one gel chamber wall of a gel chamber (12) with an access opening (14) for filling the gel chamber (12) with hydrogel, at least one fluid chamber wall of a first fluid chamber (16) for filling with fluid which has at least one biological cell, at least one first channel wall of a first channel (18) which connects the first fluid chamber (16) and the gel chamber (12) in a fluid-communicating manner, and at least one second channel

[0200] Wall of a second channel (20) which is fluidly connected to the gel chamber (12), wherein the gel chamber (12) fluidly connects the first channel (18) to the second channel (20), and a base part (54) which has at least a third channel wall of the first channel (18) and at least a fourth channel wall of the second channel (20); connecting (104) the chamber part (52) to the base part (54) such that the first channel wall is arranged opposite the third channel wall and the second channel wall is arranged opposite the fourth channel wall.

[0201] 11. Method (100) according to the preceding embodiment, characterized in that the method (100) further comprises at least the following step:

[0202] Inserting (106) at least one wire-like body between the chamber part (52) and the base part (54) such that the wire-like body extends at least between the first chamber wall and the third chamber wall and the second chamber wall and the fourth chamber wall; and / or - Attaching (108) at least one first layer (32), which has at least one first hole (38), the edge of which delimits a first portion of the gel chamber (12), and at least one second hole (40, 42), the edge of which delimits a first portion of the fluid chamber, to a base body (28) comprising at least one chamber wall of the gel chamber (12) such that the chamber wall continues at least a part of the edge of the first hole (38), to provide the chamber part (52); and / or

[0203] Attaching (110) at least one third layer (36) to a base body, which has at least one fourth hole (46) which has at least a third section of the gel chamber (12), wherein the method (100) preferably further comprises at least the following step: - Attaching (112) at least one second layer (34), which has at least one third hole (44), the edge of which delimits at least the first channel (18), the second channel (20) and a second section of the gel chamber (12), to the first layer (32) in such a way that the first section of the gel chamber (12) is fluidly connected to the second section of the gel chamber (12) and / or to the third layer (36) in such a way that the second section of the gel chamber (12) is fluidly connected to the third section of the gel chamber (12), wherein the chamber part (52) and the base part (54) are preferably connected by means of the second layer (34) be connected.

[0204] 12. Method (100) according to embodiment 10 or 11, characterized in that the method (100) further comprises at least one of the following steps: producing (114) the chamber part (52) and / or the base part (54) by means of an additive manufacturing process, preferably a 3D printing process, a subtractive manufacturing process, in particular milling, laser and / or water jet cutting, or a casting process, preferably an injection molding process; and / or

[0205] Producing (116) the first hole (38), the second hole (40, 42), the third hole (44) and / or the fourth hole (46) by a cutting process, preferably laser cutting, by an etching process and / or by a punching process.

[0206] 13. The method (100) according to any one of embodiments 10 to 12, characterized in that the second layer (34) is attached to the third layer (36) and / or the first layer (32) is attached to the second layer (34) by lamination. 14. The method (100) according to any one of embodiments 10 to 13, characterized in that the method (100) further comprises at least the following step: attaching (118) the third layer (36) to a base plate, after attaching the second layer (34) to the third layer (36); and / or that the method (100) further comprises, after fastening the base body (28) on the first layer (32), at least one of the following steps: inserting (120) an insert piece (60) having a support region (64) for supporting a hydrogel (58) which can extend into the hydrogel (58), and a holding region (62) for holding the

[0207] Inserting an insert piece (60), which can extend outside the hydrogel (58), into the gel chamber (12) such that the support region (64) extends at least into the second section of the gel chamber (12); and / or filling (122) at least one gel chamber (12) with a hydrogel (58) such that the hydrogel (58) encloses the wire-like body (56) at least within the gel chamber (12); and removing (124) the wire-like body (56) from the gel chamber (12) and the first channel (18) and / or the second channel (20). Method (100) according to embodiment 14, characterized in that the method (100) further comprises at least the following steps:

[0208] Filling (126) at least one first fluid chamber (16) with a fluid (68) comprising at least one biological cell;

[0209] Tilting (128) the device (10) and / or the microtiter plate such that the fluid (68) flows from the first fluid chamber (16) through the first channel (18) to the hydrogel (58).

Claims

Claims 1. Device (10) for producing three-dimensional cell structures in a hydrogel, comprising at least one gel chamber (12) with an access opening (14) for filling the gel chamber (12) with hydrogel, characterized in that the device (10) further comprises at least one first fluid chamber (16) for filling with fluid which has at least one biological cell, at least one first channel (18) which connects the first fluid chamber (16) and the gel chamber (12) in a fluid-communicating manner, and at least one second channel (20) which is fluid-communicatingly connected to the gel chamber (12), wherein the gel chamber (12) fluid-communicatingly connects the first channel (18) to the second channel (20).

2. Device (10) according to claim 1, characterized in that the second channel (20) connects the gel chamber (12) to the first fluid chamber (16) in a fluid-communicating manner or that the second channel (20) connects the gel chamber (12) to a second fluid chamber (22) of the device (10) in a fluid-communicating manner.

3. Device (10) according to one of the preceding claims, characterized in that the first channel (18) opens into the gel chamber (12) between the access opening (14) and at least one first bottom section (24) of the gel chamber (12) opposite the access opening (14) and / or the second channel (20) opens into the gel chamber (12) between the access opening (14) and at least one second bottom section (26) of the gel chamber (12) opposite the access opening (14).

4. Device (10) according to one of the preceding claims, characterized in that the first channel (18) and / or the second channel (20) is designed as a microchannel, wherein the microchannel has a diameter in the range between 0.1 pm and 1000 pm, preferably between 1 pm and 1000 pm, more preferably between 20 pm and 500 pm, most preferably between 50 pm and 250 pm, and / or that the gel chamber (12) has a cylindrical shape.

5. Device (10) according to one of the preceding claims, characterized in that the device (10) further comprises a wire-like element which extends through the gel chamber (12) and is removable from the gel chamber (12) and is preferably designed to be deformable.

6. Device (10) according to claim 5, characterized in that the wire-like element has a smaller diameter than the first channel (18) and / or second channel (20) and is movably arranged in the first channel (18) and / or the second channel (20).

7. Device (10) according to claim 5 or 6, characterized in that the wire-like element extends between the first channel (18) and the second channel (20).

8. Device (10) according to one of claims 5 to 7, characterized in that the wire-like element has a surface with a polyamide material.

9. Device (10) according to one of the preceding claims, characterized in that the gel chamber (12) is filled with a hydrogel, wherein the hydrogel has at least one through-opening which is fluidically connected to the first channel (18) and / or the second channel (20) and / or the gel chamber (12) preferably has at least one insert piece which has a support area for supporting the Hydrogel, which extends into the hydrogel, and a holding region for holding the insert, which preferably extends outside the hydrogel, and / or the hydrogel further preferably has at least one biological cell at least at an edge region of the passage opening.

10. Device (10) according to one of the preceding claims, characterized in that at least the first fluid chamber (16) contains a fluid which has at least one biological cell, wherein the first fluid chamber (16) preferably has at least one inlet and / or outlet opening for filling and / or draining fluid.

11. Device (10) according to one of the preceding claims, characterized in that the device (10) has at least one base body (28) and a base body (30), wherein the base body (28) extends between the access opening (14) and the base body (30), wherein the base body (28) preferably has at least one chamber wall section of the gel chamber (12) and / or the fluid chamber and / or the device (10) preferably has at least one layer which has at least one hole, the edge of which delimits the first channel (18) and / or the second channel (20) and / or a section of the gel chamber (12) and / or a section of the fluid chamber, wherein the at least one layer preferably comprises a film material, wherein the base body (30) further preferably has a base plate,wherein the at least one layer is arranged between the base plate and the base body (28) and / or the base plate is further preferably transparent at least in some areas and / or the base plate comprises a glass material and / or a plastic material, preferably a microscope slide and / or a microscope cover glass, and / or at least one first layer (32) between the base body (28) and the first channel, (18) and / or the second channel (20) and has at least one first hole (38), the edge of which delimits a first section of the gel chamber (12), and at least one second hole (40, 42), the edge of which delimits a first section of the fluid chamber, wherein the first layer (32) is preferably arranged between the base body (28) and at least one second layer (34), and the second layer (34) has at least one third hole (44), the edge of which delimits at least the first channel (18), the second channel (20), and a second section of the gel chamber (12), wherein furthermore the second layer (34) is preferably arranged between the first (32) layer and at least one third layer (36), and the third layer (36) has at least one fourth hole (46) which defines at least a third section of the gel chamber (12).

12. Microsystem (66) or microfluidic cell culture platform, which is designed in particular as a microtiter plate or slide, comprising a plurality of devices (10) according to one of the preceding claims, wherein the access openings (14) of the devices (10) are arranged next to one another in a matrix-like manner, wherein the microsystem and / or microfluidic cell culture platform preferably has a number of devices (10) in the range from 2 to 1152, preferably 2, 4, 8, 16, 32, 60, 128, 512 or 1152, most preferably 128 or 512.

13. Method (100) for producing a device (10) according to one of claims 1 to 11 or a microsystem or a microfluidic cell culture platform according to the preceding claim, comprising at least the following steps: Providing (102) a chamber part (52) which has at least one gel chamber wall of a gel chamber (12) with an access opening (14) for filling the gel chamber (12) with hydrogel, at least a fluid chamber wall of a first fluid chamber (16) for filling with fluid containing at least one biological cell, at least one first channel wall of a first channel (18) fluidly connecting the first fluid chamber (16) and the gel chamber (12), and at least one second channel wall of a second channel (20) fluidly connecting the gel chamber (12), wherein the gel chamber (12) fluidly connects the first channel (18) to the second channel (20), and a base part (54) having at least one third channel wall of the first channel (18) and at least one fourth channel wall of the second channel (20); connecting (104) the chamber part (52) to the base part (54) such that the first channel wall is arranged opposite the third channel wall and the second channel wall is arranged opposite the fourth channel wall.

14. The method (100) according to the preceding claim, characterized in that the method (100) further comprises at least the following step: inserting (106) at least one wire-like body between the chamber part (52) and the base part (54) such that the wire-like body extends at least between the first chamber wall and the third chamber wall and the second chamber wall and the fourth chamber wall; and / or attaching (108) at least one first layer (32), which has at least one first hole (38), the edge of which delimits a first portion of the gel chamber (12), and at least one second hole (40, 42), the edge of which delimits a first portion of the fluid chamber, to a base body (28) comprising at least one chamber wall of the gel chamber (12) such that the chamber wall continues at least part of the edge of the first hole (38), to provide the chamber part (52); and / or Attaching (110) at least a third layer (36) to a Base body having at least a fourth hole (46) which defines at least a third section of the gel chamber (12), wherein the method (100) preferably further comprises at least the following step: attaching (112) at least one second layer (34) having at least a third hole (44), the edge of which delimits at least the first channel (18), the second channel (20) and a second section of the gel chamber (12), to the first layer (32) in such a way that the first section of the gel chamber (12) is fluidly connected to the second section of the gel chamber (12) and / or to the third layer (36) in such a way that the second section of the gel chamber (12) is fluidly connected to the third section of the gel chamber (12), wherein the chamber part (52) and the base part (54) are preferably connected by means of the second layer (34).

15. The method (100) according to claim 13 or 14, characterized in that the method (100) further comprises at least one of the following steps: producing (114) the chamber part (52) and / or the base part (54) by means of an additive manufacturing method, preferably a 3D printing method, a subtractive manufacturing method, in particular milling, laser and / or water jet cutting, or a casting method, preferably an injection molding method; and / or producing (116) the first hole (38), the second hole (40, 42), the third hole (44) and / or the fourth hole (46) by a cutting method, preferably laser cutting, by an etching method and / or by a punching method.

16. Method (100) according to one of claims 13 to 15, characterized in that the fixing of the second layer (34) on the third layer (36) and / or the attachment of the first layer (32) to the second layer (34) is carried out by lamination.

17. Method (100) according to one of claims 13 to 15, characterized in that the method (100) further comprises at least the following step: Fastening (118) the third layer (36) to a base plate after fastening the second layer (34) to the third layer (36); and / or that the method (100) further comprises at least one of the following steps after fastening the base body (28) to the first layer (32): inserting (120) an insert (60) having a support region (64) for supporting a hydrogel (58), which can extend into the hydrogel (58), and a holding region (62) for holding the insert (60), which can extend outside the hydrogel (58), into the gel chamber (12) such that the support region (64) extends at least into the second section of the gel chamber (12); and / or filling (122) at least one gel chamber (12) with a hydrogel (58) such that the hydrogel (58) encloses the wire-like body (56) at least within the gel chamber (12);and removing (124) the wire-like body (56) from the gel chamber (12) and the first channel (18) and / or the second channel (20); 18. Method (100) according to claim 17, characterized in that the method (100) further comprises at least the following steps: Filling (126) at least one first fluid chamber (16) with a fluid (68) comprising at least one biological cell; Tilting (128) the device (10) and / or the microtiter plate such that the fluid (68) flows from the first fluid chamber (16) through the first channel (18) to the hydrogel (58).

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