System for 3D cell culture

Abstract

Fluid supply module for a cell culture system for 3D in vitro perfusion cell culture, wherein the fluid supply module comprises a fluid drive mechanism configured to move the fluid over a cell culture area of ​​a cell culture module, and wherein the fluid supply module is configured to be assembled with the cell culture module to form a cell culture system for 3D in vitro perfusion cell culture that is fluidically isolated from the external environment.

Description

[0001] The present invention relates to a system for 3D in vitro perfusion cell culture.

[0002] With the rapid advancement of 3D cell culture models, our understanding of complex human diseases, particularly cancer, is changing. Unlike conventional 2D cultures, which lack the structural and functional complexity of in vivo tissue, 3D models mimic the physiological environment much more accurately and provide better insights into cell behavior, tissue mechanics, and disease progression.

[0003] To fully exploit the potential of 3D models, 3D in vitro perfusion cell culture systems, especially dynamic in vitro perfusion plates, are gaining importance as a key technology. These systems, with microfluidic channels and controlled fluid flow, lend in vitro studies a new level of realism by enabling continuous nutrient exchange, waste removal, and the simulation of physiological forces.

[0004] Furthermore, they allow the targeted introduction of immune cells for the investigation of immunological questions and immunotherapies, and permit the study of metastasis models.

[0005] Conventional 3D in vitro perfusion cell culture systems generally consist of a cell culture well plate, with each of the wells, or a desired subset thereof, equipped with a tube so that fluid (e.g. cell culture medium) can be supplied to the well via a pump.

[0006] Peristaltic and pneumatic pumps used in conventional in vitro perfusion cell culture systems have significant disadvantages in terms of user-friendliness and cost.

[0007] Peristaltic pumps, for example, are relatively easy to operate, but require frequent maintenance due to wear and tear on the tubing. Their construction is time-consuming, and the tubing needs to be replaced regularly, which complicates handling during operation.

[0008] Pneumatic pumps, on the other hand, often comprise more complex systems with additional components such as air compressors and pressure regulators, making setup and fine-tuning more complicated. This complexity can make it difficult for users to learn how to operate them and less user-friendly for routine applications.

[0009] In terms of cost, both pump types are relatively expensive to purchase and maintain. Pneumatic pumps, in particular, require specialized equipment and infrastructure, such as compressed air sources and sophisticated control systems, which significantly increases both the initial investment and ongoing costs.

[0010] Furthermore, in conventional in-vitro perfusion cell culture systems, consumables such as tubing (in the case of peristaltic pumps) or filters and controllers (in the case of pneumatic systems) contribute further to the overall costs, making these systems less affordable for many research laboratories.

[0011] It is also important to note that conventional in vitro perfusion cell culture systems, regardless of whether a pneumatic or peristaltic pump is used, make it difficult to scale up experiments.

[0012] If a perfusion plate system requires peristaltic pumps, for example, each well needs its own tubing. Even a simple experimental setup with n=3 per experimental condition and 2 conditions therefore requires 6 tubes.

[0013] Therefore, either a special incubator would have to be developed for this purpose, or 6 very long tubes would have to be inserted into and removed from a conventional universal laboratory incubator.

[0014] Larger experimental setups with slightly more than 48 wells would require 48 tubes in a conventional in-vitro perfusion cell culture system.

[0015] In addition to the limitations mentioned above, such as changing and sterilizing the tubing, this increases the overall complexity and makes it more difficult to establish perfusion plates for everyday use.

[0016] Against this background, the present invention aims to reduce or even completely eliminate the disadvantages of the prior art.

[0017] In particular, the present invention addresses the technical problem of providing a cost-effective, efficient and user-friendly solution for 3D in vitro perfusion cell culture.

[0018] This problem is solved by the subject matter of the independent claims. Advantageous embodiments of the invention are the subject matter of the dependent claims.

[0019] A first aspect of the present invention relates to a fluid supply module for a cell culture system for 3D in vitro perfusion cell culture, wherein the fluid supply module comprises a fluid drive mechanism configured to move the fluid over a cell culture area of ​​a cell culture module, and wherein the fluid supply module is designed to be assembled with the cell culture module to form a cell culture system for 3D in vitro perfusion cell culture that is fluidically separated from the external environment.

[0020] Thus, a core concept of the present invention is to integrate the fluid supply into a cell culture system for 3D in vitro perfusion cell culture, instead of relying on external fluid sources and external tubing to direct the fluid from a pump to the cell culture area.

[0021] Instead, the cell culture system according to the present invention preferably includes its own liquid supply infrastructure (realized in the liquid supply module) and a cell culture area (realized in the cell culture module). The liquid supply module and the cell culture module are preferably detachably connected to one another, e.g., by stacking them, and thus form a portable cell culture system that can be placed in a conventional general-purpose incubator.

[0022] Due to the presence of the fluid supply module, the cell culture system according to the present invention is preferably independent of external fluid sources or pumps and can preferably be handled as a single unit. This makes it very easy to place the cell culture system in and out of the incubator without having to connect or disconnect any fluid lines.

[0023] In this sense, a cell culture system of the present invention can be considered as fluidically isolated from the external environment or “stand-alone”, since the system does not rely on an external fluid supply with pumps and hoses to pump fluid into the system.

[0024] In other words, a key element of a cell culture system of the present invention is the base plate (an example of a liquid supply module) of the cell culture system according to the present invention, as shown in the Fig. 1 and Fig. 2 shown.

[0025] A fluid supply module and / or cell culture system of the present invention offers advantages over conventional perfusion systems that rely on complex external devices such as perfusion and pneumatic pumps.

[0026] In contrast to these complex systems, the self-contained fluid supply modules and / or cell culture systems of the present invention are designed to operate fluidically independently. This eliminates the need for extensive tubing, large pumps, and complicated setups, which are both costly and time-consuming.

[0027] In other words, a fluid supply module and / or cell culture system of the present invention can be regarded as a fluidically closed or self-contained system.

[0028] This simplicity increases the accessibility and user-friendliness of the systems. Researchers can thus set up experiments quickly and reduce the risk of technical errors associated with complex equipment.

[0029] Furthermore, the standalone system according to the present invention is more compact and easier to transport than conventional systems. It can be more easily integrated into standard laboratory incubators and is better suited for high-throughput screenings.

[0030] Furthermore, its self-contained design minimizes potential contamination risks, as the entire system is isolated within a single plate and is therefore only minimally exposed to the outside environment.

[0031] Overall, a fluid supply module and / or cell culture system of the present invention offers a simplified, cost-effective and user-friendly alternative that meets the requirements of modern 3D cell culture applications, especially in daily use, high-content screening and drug discovery.

[0032] According to one embodiment, in a fluid supply module according to the present invention, the fluid drive mechanism is designed to temporarily close a fluid channel which is at least partially formed by a flexible material in order to generate a unidirectional fluid flow through the fluid channel and / or over the cell culture area.

[0033] A non-restrictive example of such a configuration is in Fig. 1 shown.

[0034] The fluid drive mechanism can include a circulating track or conveyor belt and a motor to drive the circulating track or conveyor belt.

[0035] The circulating track or conveyor belt can be equipped with at least one closing element that temporarily closes the fluid channel when the circulating track or conveyor belt is moved.

[0036] The at least one locking element can be arranged on the circulating track or conveyor belt via a flexible element, preferably a spring, thereby allowing the locking element a certain degree of mobility relative to the circulating track or conveyor belt.

[0037] The closure element may have a geometry that is at least partially rounded to ensure a secure closure without damaging the flexible material (e.g., a membrane).

[0038] The fluid supply module can further include a motor that drives the circulating track or conveyor belt, and at least one guide element that maintains the circulating track or conveyor belt at a constant, desired tension. For example, at least two guide elements in the form of rollers can be arranged at two opposite ends of the circulating track or conveyor belt to maintain tension on the belt.

[0039] According to another embodiment, the fluid drive mechanism can include a screw conveyor, preferably an Archimedean screw, which moves fluid portions to generate a unidirectional fluid flow over the cell culture area.

[0040] A non-restrictive example of such a configuration is in Fig. 2 shown.

[0041] The fluid supply module may further include a motor and preferably a ball bearing and / or a gearbox for driving the screw of the screw conveyor. However, the ball bearing and / or gearbox are optional.

[0042] The screw of the screw conveyor can have adjustable wedge structures within its grooves, thus enabling an adjustment of the fluid conveying dynamics of the screw.

[0043] The geometry of the screw can be chosen as desired to achieve the desired fluid conveying properties of the screw conveyor.

[0044] In general, a fluid supply module according to the present invention can further comprise a control unit that controls the fluid drive mechanism (e.g., the conveyor belt or the Archimedes screw) to achieve a desired flow rate and direction. A user can specify the desired flow rate and direction or program the control unit to execute a desired culture protocol.

[0045] A liquid supply module according to the present invention is preferably designed as a reusable (preferably autoclavable) module. The liquid supply module can be used with a disposable cell culture module for single use.

[0046] A fluid supply module according to the present invention can be configured to be combined with a cell culture module in the form of a conventional cell culture plate, in particular a single-well plate or multi-well plate, to form a cell culture system for 3D in vitro perfusion cell culture, which is fluidically separated from the external environment.

[0047] This makes it possible to use a liquid supply module according to the present invention with conventional cell culture plates.

[0048] In general, within the scope of the present invention, the liquid supply module and the cell culture module can either be designed as two separable modules or as a single component.

[0049] The liquid supply module can be assembled into the cell culture system for 3D in vitro perfusion cell culture by stacking the cell culture plate on top of the liquid supply module with the conventional cell culture plate.

[0050] The simple stacking of the cell culture module onto the fluid supply module to form the 3D in vitro perfusion cell culture system results in a particularly compact and easy-to-assemble cell culture system.

[0051] Another aspect of the present invention relates to a cell culture system for 3D in vitro perfusion cell culture, comprising a fluid supply module according to the present invention and a cell culture module, preferably in the form of a conventional cell culture plate, in particular a single-well or multi-well plate. The fluid supply module and the cell culture module are preferably detachably connected to each other or designed as a single component.

[0052] The cell culture system is preferably fluidically separated from the external environment and / or independent of any external fluid source.

[0053] The cell culture system is preferably portable and, furthermore, preferably designed so that it can be used in any conventional cell culture incubator that does not have fluid lines to supply the cell culture system with fluid.

[0054] Since the cell culture system does not need to be connected to an external fluid supply via, for example, hoses, the cell culture system according to the present invention, preferably the entire system, is designed as a monolithic block, preferably in the form of a rectangular cuboid.

[0055] In other words, unlike a conventional cell culture system with a more distributed structure and a multitude of external fluidic auxiliary devices such as tubing and / or pumps, the cell culture system according to the present invention can be designed as a single “block” that can be easily moved with just one handle.

[0056] In a cell culture system according to the present invention, the cell culture module is preferably designed as a disposable module for single use.

[0057] A cell culture system according to the present invention can further comprise a flexible material, preferably in the form of a flexible membrane, which separates the cell culture module from the fluid supply module and at least partially forms a fluid channel that can be temporarily closed by the fluid drive mechanism of the fluid supply module.

[0058] Such a structure is particularly advantageous when a circulating belt or conveyor belt, as described above, is used for fluid supply. The flexible membrane can be part of the cell culture module, as exemplified in Fig. 1 shown.

[0059] Furthermore, a cell culture system according to the present invention can have at least two openings, preferably each containing a fluid-permeable barrier made of a porous material or a membrane, so that the fluid supplied by the fluid supply module can enter the cell culture area of ​​the cell culture module.

[0060] Thus, a cell culture system according to the present invention can be used for cultivating particularly sensitive cells. In conventional cell culture systems, fluid is often introduced directly into a cell culture well via a tube. The flow of the fluid causes shear stress on the cells.

[0061] In a cell culture system according to the present invention, the fluid can reach the cells in a more indirect and controlled manner by passing through the fluid-permeable barriers (preferably made of a porous material or a membrane). This reduces the shear stress on the cells.

[0062] Another aspect of the present invention relates to the use of a fluid supply module according to the present invention for 3D in vitro perfusion cell culture, namely for supplying fluid to a cell culture area of ​​a cell culture module.

[0063] The fluid supply module is preferably part of a cell culture system according to the present invention.

[0064] Naturally, all features described in connection with the liquid supply module and the cell culture system of the present invention also apply to the aforementioned use of the liquid supply module.

[0065] In other words, the features of the present invention can be summarized as follows.

[0066] The present invention relates to a self-contained dynamic perfusion plate system (an example of a cell culture system) for in vitro 3D cell culture. It comprises an upper plate (an example of a cell culture module) with porous recesses (an example of openings with fluid-permeable barriers), a fluid transport mechanism (an example of a fluid supply module), and a motorized drive unit for controlling the fluid movement across the culture surface without the need for external tubing or pumps.

[0067] The perfusion plate system can be designed to replicate physiological perfusion conditions within an in vitro 3D cell culture model. The system's compact, modular structure, which contains the fluid flow within a self-contained unit, facilitates integration into standard laboratory incubators.

[0068] According to one embodiment of the perfusion plate system, the fluid transport mechanism comprises a circumferential track with attached wedges (an example of closure elements). When driven by motorized rotary devices, these wedges press against an elastic material and create fluid pockets, thereby enabling controlled and uniform fluid movement across the growing area.

[0069] This embodiment is exemplified in Fig. 1 shown.

[0070] The circulating track may incorporate a series of belt guides (an example of guide elements) and mechanisms positioned along the base plate (an example of a liquid supply module) to maintain consistent belt tension and support uninterrupted liquid flow over extended culture periods.

[0071] The perfusion plate system may further include a fluid drive (which may include a control unit), preferably arranged at the base of the rotating track, to control the flow rate and flow direction, thus providing adaptable perfusion conditions for different cell culture requirements.

[0072] According to another embodiment of the perfusion plate system, the fluid transport mechanism includes an Archimedean screw below the upper plate. This screw traps fluid in its helical grooves and, as it rotates, moves it across the culture surface, thereby simulating physiologically relevant flow patterns.

[0073] This embodiment is exemplified in Fig. 2 shown.

[0074] The Archimedean screw can be driven by a gearbox 8 and a motor 9. Ball bearings 10 ensure low-friction rotation, enabling precise control of fluid supply and nutrient gradients in the growing environment.

[0075] The Archimedean screw can have adjustable wedge structures in its grooves, allowing the fluid transport dynamics to be adapted as needed.

[0076] In a perfusion plate system according to the present invention, the upper plate (example of a cell culture module) can be designed as a disposable component, while the lower plate (example of a fluid supply module), which contains the motor and the fluid transport mechanism, is reusable. This ensures cost-efficiency and sterility for repeated use in the laboratory.

[0077] The perfusion plate system can also be configured to support high-throughput screening applications. Its self-contained design minimizes the risk of contamination and simplifies setup for routine use in drug discovery, cancer research, and tissue-specific studies.

[0078] It should be noted here that the terms "ein" and "eine" do not necessarily mean exactly one element—although this is a possible interpretation—but can also encompass several elements. Likewise, the use of the plural includes the presence of the element in the singular, and conversely, the singular also includes multiple instances of the element in question.

[0079] Furthermore, all features of the invention described herein can be combined with one another as desired or, as required, claimed individually.

[0080] Further advantages, features, and effects of the present invention will become apparent from the following description of preferred embodiments with reference to the figures. The figures show: Fig. 1: a cell culture system according to a first embodiment of the invention, and Fig. 2: a cell culture system according to a second embodiment of the invention.

[0081] The in Fig. The cell culture system shown is designed to enable controlled fluid movement across a 3D cell culture environment. It utilizes a circulating belt mechanism 3 that dynamically regulates the flow of nutrients and waste products.

[0082] This design comprises a top plate with porous depressions 1 through which fluid can pass and come into direct contact with the cell cultures. Below the top plate, an elastic material 2 forms a flexible interface that adapts to the movement of a series of wedges (examples of closure elements) mounted on the surrounding band 3.

[0083] These wedges are strategically positioned along the belt to create separate fluid pockets by pressing against the elastic material 2 (membrane). Each pocket is moved along the plate by motor rotators 4, which drive the belt 3 in an endless loop, thus enabling a continuous and uniform fluid distribution.

[0084] The base plate contains belt guides 5 and spring mechanisms that provide structural support and help maintain constant belt tension to ensure smooth and uninterrupted operation. The wedges may each have a spring 6 at their base.

[0085] In addition, a fluid drive system 7 at the base drives the entire system and enables precise control of flow rate and flow direction.

[0086] The compact, self-contained design of this system eliminates the need for external hoses or pumps, significantly reduces the risk of contamination, and is compatible with common laboratory incubators.

[0087] By replicating natural perfusion dynamics, this system creates more physiologically relevant culture conditions - especially for studies that require continuous nutrient exchange and waste removal.

[0088] The in Fig. The configuration shown is based on a circumferential band with wedges that press against an elastic material to form fluid pockets that can be moved across the plate.

[0089] Various wedge designs are possible. Separate gears can be used to rotate the wedges that press against the elastic material or membrane.

[0090] The embodiment of the Fig. 2 uses an Archimedean screw mechanism 11 to facilitate fluid movement across the 3D cell culture environment. This utilizes the principle of screw-based fluid transport.

[0091] In this configuration, the upper plate contains porous depressions that allow direct fluid exchange with the cultured cells.

[0092] A rotating Archimedean screw 11 captures the fluid in controlled portions. As it rotates, the screw's spiral grooves create a series of small fluid chambers. Each groove of the screw 11 is designed to trap fluid and transport it forward with the rotation, thus achieving a uniform, steady flow across the cell culture area.

[0093] The base plate houses a gearbox 8 and a motor 9, which drive the Archimedean screw 11, as well as ball bearings 10, which ensure low-friction, smooth movement.

[0094] By adjusting the motor speed, the flow rate and fluid supply pattern can be finely controlled. This allows for the precise simulation of nutrient gradients and waste removal over extended cultivation periods. The gearbox 8 and / or the ball bearings 10 are not strictly necessary due to the indirect magnetic drive.

[0095] The fluid conveyed by the Archimedean screw 11 flows through an opening with a porous barrier 12 to reach the cell culture area of ​​the upper plate.

[0096] Furthermore, the screw design can be modified to integrate different wedge patterns, allowing the fluid dynamics to be adapted to specific requirements.

[0097] This system is highly modular and offers the possibility of combining a disposable top plate with a reusable bottom plate that can be autoclaved to maintain sterility.

[0098] Similar to the embodiment of the Fig. 1. This self-contained setup also requires no external tubing or pumps. This increases compatibility with conventional laboratory equipment and reduces setup complexity.

[0099] The screw design 11 enables a sustained, physiologically relevant fluid flow. This makes it particularly suitable for high-throughput screenings as well as applications in cancer research, drug testing, and studies of tissue-specific responses under dynamic conditions.

[0100] Various wedge configurations can be used on the screw. Furthermore, instead of separate upper (single-use) and lower (autoclavable) plates, it is conceivable to use a single (disposable) plate that integrates the functions of both the upper and lower plates.

[0101] The in the Fig. 1 and Fig. The two cell culture systems shown are self-contained, dynamic in-vitro perfusion systems designed for 3D cell culture applications.

[0102] By integrating microfluidic channels, a motorized fluid drive, and user-friendly modularity, these systems enable a continuous flow of media to simulate, for example, controlled gradients of signaling molecules, mechanical forces, and the remodeling of the extracellular matrix (ECM). This provides insights into how diseased cells adapt and proliferate in different tissue environments.

[0103] Such systems also allow researchers to investigate interactions between organs – for example, in metastasis studies, where they model how tumor cells from a primary tumor interact with distant organ environments, as well as the inclusion of immune cells. The simplicity of these systems eliminates the need for external pumps, thus increasing accessibility and ease of use in everyday laboratory practice.

[0104] Thus, the following advantages can be achieved with a cell culture system for 3D in vitro perfusion cell culture - in particular according to the present invention: Increased physiological relevance: Dynamic in vitro perfusion plates offer a significant advantage because they replicate key aspects of tissue-level perfusion as they occur in the human body. These plates allow for the simulation of blood flow and interstitial fluid movement, which are crucial for maintaining cell viability and functionality over extended culture periods. This continuous flow more accurately mimics the in vivo microenvironment than static cultures, allowing cells in 3D models to experience realistic gradients of oxygen, nutrients, and growth factors. Consequently, perfusion plates enable more physiologically relevant investigations of tissue-specific phenomena such as tumor progression, fibrosis, and immune cell infiltration, making them ideal for translational research.

[0105] Improved Investigation of Tumor Microenvironments: Perfusion plates are particularly useful for cancer research because tumor microenvironments play a central role in disease progression and treatment response. Tumors in vivo are influenced by dynamic interactions between cancer cells, stromal cells, immune cells, and the ECM—all within a constantly changing biochemical and biomechanical environment. Dynamic in vitro perfusion systems can simulate these interactions by allowing controlled gradients of signaling molecules, mechanical forces, and ECM remodeling. This provides insights into how cancer cells adapt and spread in different tissue environments. Such systems also allow researchers to investigate organ-organ communication in metastasis studies by modeling how tumor cells from a primary tumor interact with distant organ environments (such as the liver or lungs).

[0106] Accelerated drug development and screening: The limitations of traditional in vitro models are well-known in drug development. Promising candidates often fail in clinical trials due to lack of efficacy or unexpected toxicity that went undetected in early tests. Dynamic perfusion plates offer a more realistic platform and can thus increase the predictive accuracy of preclinical drug screenings. They support high-content screening methods that allow real-time assessments of cellular responses under physiologically relevant conditions (e.g., drug concentration gradients and prolonged exposure). This can significantly reduce the failure rate in drug development, as effective therapies can be identified more accurately before being transferred to costly in vivo or clinical trials.

[0107] Improved research into fibrosis and inflammatory diseases: Dynamic perfusion plates offer valuable applications beyond cancer research in the investigation of fibrosis and inflammatory diseases. Fibrosis is a key feature of numerous chronic diseases and is characterized by excessive ECM deposition and tissue stiffness. Static 3D cultures often fail to adequately represent the dynamic remodeling processes of fibrosis. Perfusion plates, on the other hand, allow a continuous flow of cytokines, ECM components, and other factors, thus facilitating studies of how cellular responses to ECM stiffness and composition change over time. This approach is particularly relevant for understanding active fibrosis in organs such as the liver, heart, and lungs, where complex cell-ECM interactions govern disease progression.

[0108] Bridging the translational gap: Dynamic perfusion systems in 3D culture models bridge the gap between preclinical in vitro research and in vivo studies, thus closing a crucial gap in translation. By more accurately replicating the complexity of living tissues, these systems reduce the discrepancy between laboratory and patient outcomes and increase the likelihood that in vitro findings will translate into clinical success. This is particularly promising for personalized medicine, as perfusion-based 3D cultures can be adapted to individual patient conditions and represent a powerful tool for testing patient-specific responses to therapies.

Claims

[1] Fluid supply module for a cell culture system for 3D in vitro perfusion cell culture, wherein the fluid supply module comprises a fluid drive mechanism configured to move the fluid over a cell culture area of ​​a cell culture module, and wherein the fluid supply module is configured to be assembled with the cell culture module to form a cell culture system for 3D in vitro perfusion cell culture which is fluidically isolated from the external environment. [2] Fluid supply module according to claim 1, wherein the fluid drive mechanism is configured to temporarily close a fluid channel which is at least partially formed by a flexible material in order to generate a unidirectional fluid flow through the fluid channel and / or over the cell culture area. [3] Fluid supply module according to claim 2, wherein the fluid drive mechanism comprises a circulating track or conveyor belt and a motor (9) for driving the circulating track or conveyor belt, wherein the circulating track or conveyor belt is equipped with at least one closing element which temporarily closes the fluid channel when the circulating track or conveyor belt is moved. [4] Liquid supply module according to claim 3, wherein the at least one closure element is arranged on the circulating track or conveyor belt via a flexible element, preferably a spring (6), thereby enabling the at least one closure element to have a certain degree of mobility relative to the circulating track or conveyor belt. [5] Liquid supply module according to claim 3 or 4, further comprising a motor (9) that drives the circulating track or conveyor belt, and at least one guide element that holds the circulating track or conveyor belt with a constant desired tension. [6] Fluid supply module according to claim 1, wherein the fluid drive mechanism comprises a screw conveyor, preferably an Archimedean screw, which moves fluid portions to generate a unidirectional fluid flow over the cell culture area. [7] Liquid supply module according to claim 6, further comprising a motor and preferably a ball bearing (10) and / or a gearbox (8) for driving the screw conveyor. [8] Fluid supply module according to claim 6 or 7, wherein the screw of the screw conveyor has adjustable wedge structures within the grooves of the screw, thereby enabling an adjustment of the fluid conveying dynamics of the screw. [9] Fluid supply module according to any of the preceding claims, further comprising a control unit configured to control the fluid drive mechanism to effect a desired flow rate and flow direction. [10] Liquid supply module according to one of the preceding claims, wherein the liquid supply module is preferably designed as an autoclavable reusable module. [11] Fluid supply module according to one of the preceding claims, wherein the fluid supply module is configured to be assembled with a cell culture module in the form of a conventional cell culture plate, in particular a single-well plate or multi-well plate, to form a cell culture system for 3D in vitro perfusion cell culture which is fluidically separated from the external environment. [12] Liquid supply module according to claim 11, wherein the cell culture plate is stacked on the liquid supply module to form the cell culture system. [13] Cell culture system for 3D in vitro perfusion cell culture, comprising a fluid supply module according to one of the preceding claims and a cell culture module, preferably in the form of a conventional cell culture plate, in particular a single-well plate or multi-well plate. [14] Cell culture system according to claim 13, wherein the cell culture system is fluidically separated from the external environment and independent of any external fluid source. [15] Cell culture system according to claim 13 or 14, wherein the cell culture system is portable and preferably designed to be used with any conventional cell culture incubator that does not have fluid lines for supplying fluid to the cell culture system. [16] Cell culture system according to any one of claims 13 to 15, wherein the cell culture system, preferably the entire cell culture system, is designed as a monolithic block, preferably in the form of a rectangular cuboid. [17] Cell culture system according to any one of claims 13 to 16, wherein the cell culture module is designed as a disposable module for single use. [18] Cell culture system according to one of claims 13 to 17, further comprising a flexible material, preferably in the form of a flexible membrane, which separates the cell culture module from the fluid supply module and at least partially forms a fluid channel which is temporarily closed by the fluid drive mechanism of the fluid supply module. [19] Cell culture system according to one of claims 13 to 18, further comprising at least two openings, each preferably containing a fluid-permeable barrier made of a porous material or a membrane, so that fluid supplied by the fluid supply module can enter the cell culture area of ​​the cell culture module. [20] Use of a fluid supply module according to one of claims 1 to 12 for 3D in vitro perfusion cell culture, namely for supplying fluid to a cell culture area of ​​a cell culture module. [21] Use according to claim 20, wherein the liquid supply module is part of a cell culture system according to any one of claims 13 to 19.

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