Autonomous microfluidic cell culture platform

Abstract

A cell culture platform for integration into detection systems, in particular microscopy systems, wherein the cell culture platform comprises: a microfluidic system; at least one cell culture component; and a drive system at least comprising a micropump, a control unit and an energy supply unit.

Description

[0001] Autonomous microfluidic cell culture platform

[0002] The invention relates to an autonomous microfluidic cell culture platform.

[0003] background

[0004] In vitro cell culture systems are essential in science and industry for investigating, for example, fundamental cellular processes, diseases, or the effectiveness of medications. Instead of the usual static conditions, cells are cultured dynamically under an applied flow to reconstruct blood flow and create physiologically relevant conditions. The flow induces shear forces that lead to changes in cellular processes such as cell differentiation, membrane organization, cell orientation, etc. This impacts, among other things, the toxicity and efficacy of substances such as medications and materials for medical devices. Dynamic cell culture systems therefore show great potential in drug testing in the pharmaceutical industry, as they better reflect the pharmacology and pharmacokinetics of an active ingredient, such as its uptake into the cell.In this way, ethically questionable animal testing can be significantly reduced and drug discovery and development can be accelerated. Furthermore, the use of human cells within dynamically cultured cell culture systems could bring a solution to the problem of limited transferability from animals to humans. This also creates immense economic benefits, as drug candidates that are ineffective in humans can be eliminated early on and the failure of drug candidates after costly clinical phases can be reduced. In addition to drug research, dynamically cultured cell culture systems can also be used for the fundamental study of diseases. For example, they can be used to better investigate and understand complex infection processes caused by viruses or bacteria that are influenced by an adjacent current.

[0005] Despite the enormous potential of dynamically cultivated cell culture systems, they are by no means established and are mainly found in scientific research as proof-of-concept systems. They typically consist of so-called microfluidic systems and require precise pumps and microscopes with special incubators for operation, which lead to complex experimental setups and require a high level of expertise in this field. Access to these systems is difficult for research groups or companies that only have expertise in general cell culture but not in the use or even development of dynamically cultivated cell culture systems.

[0006] Monitoring and analysis of cell culture within cell culture systems is typically performed using standard microscopes in combination with top-mounted or on-stage incubators. For traditional, static cell culture in tissue culture plates in well-plate format and in the absence of flow, so-called live-cell imaging systems ("real-time cell imaging systems") are increasingly being used. These systems enable fully automated, continuous monitoring of cells, thus accelerating workflows and significantly increasing data yield and knowledge acquisition. Numerous different commercial systems are available, ranging from inexpensive light microscopes to expensive confocal fluorescence microscopes. All systems have in common that they can be operated either in a conventional incubator or serve as incubators themselves.However, the integration of dynamic cell culture systems into live-cell imaging systems is a major challenge, as tubes or cables connecting the cell culture system to peripherals such as pumps and control electronics are usually difficult or even impossible to integrate into live-cell imaging systems.

[0007] The present invention is therefore based on the object of providing an autonomously usable, portable cell culture platform for dynamic cell cultivation that is compatible with common microscopy systems.

[0008] Summary of the invention

[0009] The solution to this problem in a cell culture platform of the type mentioned above consists in particular in that the cell culture platform comprises: a microfluidic system, a cell culture component and a drive system comprising at least one micropump, a control unit and a power supply unit. The invention describes a preferably rechargeable or battery-operated and thus portable cell culture platform, the external dimensions of which are based on a standardized tissue culture plate in the so-called well-plate format and can thus be operated within live cell imaging systems. The system consists of a microfluidic chip which contains access points for, for example, four cell culture components. Each platform contains at least one media circuit, which is operated separately by a micropump which, as part of the drive system, is located directly on the microfluidic chip.The pumps are operated independently via a suitable controller and a replaceable battery, eliminating the need for tubing and cables. Unlike commercially available cell culture platforms, the platform allows for use in commercial live-cell imaging systems.

[0010] Preferably, the microfluidic system, the at least one cell culture component, and the drive system are coupled to the cell culture platform in a form-fitting and / or force-fitting manner. A form-fitting coupling describes, in particular, that all components are functional components. A force-fitting coupling has the advantage that the individual components can be retrofitted or plugged in. A form-fitting and force-fitting coupling occurs, for example, when the platform is first cast during production and then, for example, the drive system is plugged onto it and then soldered.

[0011] Preferably, the dimensions of the cell culture platform correspond to the size standard according to the ANSI / SBS 1-2004 standard for microtiter plates. In particular, the cell culture platform has a length of 120-130 mm, a width of 80-90 mm, and a height of 50 mm. In one embodiment, these dimensions are maximum dimensions. This makes the cell culture platform compatible with standardized systems, such as automated imaging systems, incubators, and robotic systems.

[0012] In one embodiment, the microfluidic system is designed to be detachably connected to the drive system. This allows, for example, the microfluidic system to be sterilized separately from the drive system. This also makes the microfluidic system interchangeable and suitable for single-use applications.

[0013] In one embodiment, the cell culture component is part of the microfluidic system, wherein, in particular, the cell culture component is a cell culture chamber with an inlet and outlet that is sealed when coupled. This enables simple fabrication of the microfluidic system together with the predefined cell culture component.

[0014] In a further embodiment, the cell culture component comprises a device, in particular a connector, for cell culture inserts. External cell culture inserts can thus be connected to the device without the need for tubes, further simplifying the autonomous operation of the cell culture platform and making it modular in its use.

[0015] In one embodiment, the cell culture platform comprises at least one medium reservoir (reservoir for cell culture medium), which is arranged, in particular, laterally adjacent to the cell culture component on the cell culture platform and / or which is arranged, in particular, vertically adjacent to the cell culture component. This increases the compactness of the system and thus its throughput and operating time.

[0016] In one embodiment, the drive system is designed such that the cell culture platform can be operated for at least 24 hours, in particular at least 72 hours, and more particularly at least 100 hours. This allows cell culture studies to be conducted uninterrupted over a long period of time. Furthermore, the battery module (in particular the first one) can be easily replaced after, for example, one day.

[0017] In one embodiment, the drive system is covered by a lid. In a further embodiment, the lid can be connected to the cell culture platform by means of a screw connection, plug connection, and / or magnetic connection. The screw connection, plug connection, and / or magnetic connection can be positively connected to the cell culture platform by a seal. This allows the drive system, in particular electronic components of the drive system, to be protected from liquids, for example, cell culture medium or rinsing medium. Furthermore, the drive system, in particular electronic components of the drive system, can be sealed against the high humidity present in incubators, for example, 95% or more, and can be protected from corrosion.

[0018] In one embodiment, the at least one micropump of the drive system is operated by direct current. This avoids the extensive electronics required for AC-powered pumps and achieves the necessary compactness.

[0019] In a further embodiment, the control electronics comprise communication modules for wireless communication with the platform. This enables wireless operation in closed imaging systems, preferably for an operating time of 1 to 3 days.

[0020] In one embodiment, the power supply comprises a replaceable and rechargeable first battery module and a second battery module permanently connected to the cell culture platform. Preferably, the first battery module has a greater storage capacity than the second battery module. Further preferably, the first battery module can be used for the long-term power supply of the system, and the second battery module can be used to operate the cell culture platform when the first battery module is replaced. This allows the time required to replace the first battery module with the second battery module to be bridged.

[0021] A second aspect of the present invention describes a method for implementation in a detection system, in particular in a microscopy system. In the method, an examination of a cell culture is carried out using an autonomously operable cell culture platform, in particular the cell culture platform described above. The method allows the examination of cell cultures in common detection and microscopy systems without the need for special accessories. This is made possible in particular by the autonomous operation of the cell culture platform.

[0022] A third aspect of the present invention describes the use of a drive system on a cell culture platform, in particular the cell culture platform described above, wherein the drive system is designed such that the cell culture platform can be operated autonomously by means of the drive system. This allows the cell culture platform to be used in common detection systems, such as microscopy systems, without the need for special, complex and often expensive connection systems or external drive systems.

[0023] In one embodiment, the drive system comprises at least one micropump, a control unit and a power supply unit.

[0024] In a further embodiment, the cell culture platform can be operated by the drive system for a running time of at least 24 hours, in particular at least 72 hours, and more particularly at least 100 hours. This enables the uninterrupted examination of cell cultures over long periods of time.

[0025] The invention is described in more detail below using several preferred embodiments.

[0026] It shows:

[0027] Fig. 1 a cell culture platform;

[0028] Fig. 2 a microfluidic system with direct integration of the cell culture component in side view;

[0029] Fig. 3 a microfluidic system with direct integration of the cell culture component in top view;

[0030] Fig. 4 a microfluidic system with indirect integration of the cell culture component in side view;

[0031] Fig. 5 a microfluidic circuit with several cell culture chambers (shown here as similar);

[0032] Fig. 6 shows a section of a cell culture platform with a medium reservoir arranged vertically to the cell culture component in a side view;

[0033] Fig. 7 shows a section of a cell culture platform with a medium reservoir arranged vertically to the cell culture component in plan view;

[0034] Fig. 8-10 Mounting options for a housing of the drive system to a cell culture platform; Fig. 11 shows a drive system on a cell culture platform.

[0035] Identical or similar features are provided with the same reference symbols.

[0036] The following examples serve to illustrate the invention without limiting its scope, but may themselves represent specific embodiments of the invention.

[0037] Cell cultures are understood to mean biological cells of any kind, including eukaryotic cells, but also microorganisms and viruses, for example.

[0038] Figure 1 shows an autonomously operable cell culture platform 100 according to the invention with eight cell culture components 104, each of which is connected to the drive system 106 via a microfluidic system 102 with, in this case, eight medium reservoirs 114. The drive system 106 comprises, by way of example, eight micropumps 108, a control unit 110, a first battery module 118, and a second battery module 120. The microfluidic system 102 connects, in particular, each of the eight cell culture components 104 to a micropump 108 via a medium reservoir 114. Each micropump 108 is communicatively connected to the control unit 110 by means of a cable connection inside the drive system 106. The two battery modules 118, 120 supply the control unit 110 and the micropumps 108 with energy.

[0039] The following describes exemplary, preferred embodiments of the individual components of the cell culture platform 100. Unless otherwise noted, all components can be combined with one another. Preferred component combinations are marked as such.

[0040] The microfluidic system 102 comprises a channel system and connects the cell culture components 104 to the medium reservoir(s) 114 and the drive system 106, in particular to the micropumps 108. The microfluidic system 102 thus comprises connection elements or interfaces to the drive system 106. Optionally, the microfluidic system 102 can additionally comprise valves and / or sensors. The microfluidic system 102 contains the entire cell culture medium for the dynamic cultivation of cells in one or more medium circuits.

[0041] The microfluidic system 102 can be manufactured, for example, by milling the microchannels, injection molding plastics, or by subsequent sealing by plastic welding. Alternatively, the microfluidic system 102 can be manufactured using lithographic processes or additive manufacturing or 3D printing.

[0042] Preferably, the microfluidic system 102 is separable from the drive system 106, so that in particular a separate sterilization of the microfluidic system 102 is possible.

[0043] The microfluidic system 102 consists of one or more types of plastic. The material is preferably biocompatible with animal cells, for example, according to ISO EN 10993 (version 2023). This enables use in animal cell culture. The microfluidic system 102 is preferably optically transparent and non-absorbent or only weakly absorbent for wavelengths between 300 nm and 900 nm, and exhibits low autofluorescence (at least in the range of the cell culture(s)) to allow examination of the cells using various microscopy techniques.

[0044] Preferably, the microfluidic system 102 is sterilizable by gamma or beta irradiation. Alternatively, the microfluidic system 102 can be sterilized by heat sterilization (dry or steam) or chemical sterilization (ethylene oxide, hydrogen peroxide, chlorine dioxide, peracetic acid, nitrogen dioxide). Combinations of several methods are possible.

[0045] Preferably, the microfluidic system 102 is implemented as a replaceable, single-use component. This enables simple and rapid deployment of the cell culture platform 100. Furthermore, the risk of damage to the cell culture platform 100 during cleaning is avoided. Furthermore, potential residues and contamination from previous cell investigations are avoided, allowing for greater reproducibility of experimental results. A channel system connects all subcomponents of the microfluidic system 102 and thus serves to distribute cell medium and additives. The channel system consists of at least one closed circuit that connects a cell culture chamber or cell culture component 104, a medium reservoir 114, and the interface(s) to a micropump 108, as shown, for example, in Figure 2 or 3.Alternatively, a circuit can also connect multiple cell culture components 104 on a chip, as shown in Figure 5. The multiple cell culture components 104 can be of the same or different types. Such a circuit serves to parallelize or enable the investigation of interactions between different cell culture components 104.

[0046] The cell culture components 104 contain the cells that are cultivated and examined under flow and the influence of shear forces. The cell culture components 104 are preferably arranged according to the dimensions of standardized cell culture plates. The positioning and arrangement of the cell culture components 104 preferably follows commercially available 6-, 12-, 24-, 48-, 96-, or 384-well plate systems. This ensures integration of the cell culture platform 100 for use in automated microscopy using standardized microscopes.

[0047] The cell culture component 104 consists of a cell culture chamber connected to one or more inlets and outlets of the channel system of the microfluidic system 102. The cell culture component 104 can be integrated directly or indirectly into the microfluidic system 102. Figures 2 and 3 show exemplary arrangements for direct integration of the cell culture component 104 as part of the microfluidic system 102 in a side view and a top view. The cell culture component 104 is configured as a sealed cell culture chamber with an inlet and outlet. This embodiment enables simple production of the microfluidic system 102 together with a predefined cell culture component 104.

[0048] In an alternative embodiment, the cell culture component 104 can be designed to be indirectly connectable to the microfluidic system 102, as shown in Figure 4. In this embodiment, the cell culture component 104 contains a connector or device for connecting external cell culture inserts. These external cell culture inserts can then be connected to the device, for example, without tubing. In this embodiment, a seal is located between the inlets and outlets of the microfluidic system 102 and the cell culture insert.

[0049] Preferably, the cell culture platform 100 comprises at least one medium reservoir 114. The medium reservoir 114 serves as an access point for adding or removing the medium from the microfluidic system 102. Such a medium reservoir 114 consists of at least one outlet and inlet for connecting to the channel system, as well as an upwardly directed opening for adding or removing the medium. The at least one medium reservoir 114 is arranged according to the dimensions of standardized cell culture plates, with arrangements of 6, 12, 24, 48, 96, or 384 cell plates being possible. This enables the automation of the medium change by pipetting robots.

[0050] The at least one medium reservoir 114 can be adjacent to the cell culture component 104 or arranged vertically above it. A vertical arrangement of a medium reservoir 114 above a cell culture component 104 is shown in Figures 6 and 7 in side and top views. Here, the bottom of the reservoir is not directly open to the cell culture component 104, but is connected to the cell culture component 104 via the channel system. In this case, the medium reservoir 114, like the cell culture component 104, fulfills all (in particular, those mentioned above) material requirements for microscopy (transparency, low autofluorescence). In this case, the medium reservoirs 114 and the cell culture components 104 can, for example, be manufactured from two different injection-molded parts and subsequently joined by plastic welding.

[0051] The drive system 106 of the cell culture platform 100 according to the invention comprises at least one micropump 108, a control unit 110, and at least one power supply unit 112, as shown in Figure 1. Additionally, the drive system 106 may contain sensors and / or valves and corresponding interfaces to these components.

[0052] The service life of the cell culture platform 100 according to the invention is at least 24

[0053] hours, preferably at least 72 hours, particularly preferably at least 100

[0054] Hours. The service life depends on the capacity of the energy supply unit 112, in particular the capacity of the first battery module 118 described in more detail below, and the energy consumption of the cell culture platform 100. The consumption of the cell culture platform 100 is primarily determined by the energy consumption of the micropumps 108. Furthermore, the control unit 110 or communication unit or other sensors, for example, can contribute to the energy consumption. The service life can be approximately determined from the product of the operating time of the energy supply unit 112 and the number of micropumps 108. Due to the compact design of all electronic components and the arrangement on the microfluidic system 102, the product of the autonomous operating time and the number of continuously and independently operated micropumps 108 can, in one embodiment, be at least 100 hours.For example, a cell culture platform 100 with four independent, continuously operating micropumps 108 can be operated for a period of at least 25 hours. This ensures a sufficiently long cultivation period within closed systems without an external power supply.

[0055] The drive system 106 additionally comprises a housing 116, which, once attached to the microfluidic system 102, protects all internal parts of the drive system 106 from moisture, as shown in Figures 8-10. The housing 116 serves to protect the electronics from corrosion caused by high humidity in an incubator. For this purpose, a seal 122 (Figure 8) is located at the contact points 124 between the housing 116 and the cell culture platform 100. Other elements of the drive system 106 that protrude through the housing 116 to the outside are sealed by gluing or caulking. Examples of such elements are buttons, displays, and / or LEDs. The housing 116 can be placed, plugged, or slid onto the cell culture platform 100, for example, using a screw connection (Figure 9) or a magnetic connection (Figure 10), or can be removed from the cell culture platform 100.This enables safe and easy attachment of the drive system 106 after sterilization of the microfluidic system 102.

[0056] The drive system 106 further comprises at least one micropump 108. Such a micropump 108 is in direct contact with the microfluidic system 102. In a preferred embodiment, the at least one micropump 108 can be operated by direct current. In this way, the extensive electronics required for AC-operated pumps are avoided and the necessary compactness is achieved. The pump(s) can induce flow within a circuit via a flexible element of the microfluidic system 102, such as integrated hoses or membranes. The micropumps 108 can be designed, for example, as peristaltic pumps, ball-bearing squeeze pumps, or diaphragm pumps. The micropumps 108 can be operated continuously and enable an uninterrupted media flow.In other words, the micropumps 108 can be operated, for example, during microscopy in detection systems, during robotic transport, or during medium exchange. As a result, the cells in the cell culture component 104 are exposed to physiological flows at all times. Stopping the flow, as is necessary in other systems, can have an unwanted negative impact on the cultivation conditions and experimental results and can be avoided in the cell culture platform 100 according to the invention.

[0057] The drive system 106 also includes a control unit 110. Such a control unit 110 includes a controller with a microprocessor and integrated memory, which acts as a computer, driver modules for controlling micropumps 108, valves and / or sensors, and communication modules for wireless communication with the cell culture platform 100. The controller ensures the processing and output of commands via firmware stored on it, as well as the control and power supply of all electronic or mechatronic components of the drive system. Communication takes place, for example, via Bluetooth or wireless LAN (WLAN) and is used to send commands to the cell culture platform 100 and to read data from the cell culture platform 100. Alternatively, a predefined program can be loaded onto the platform, started, and executed, or the data can be subsequently downloaded from the platform.

[0058] The drive system 106 comprises a power supply unit 112. The power supply unit 112 consists of at least one (preferably separable) energy storage device that supplies the electronic components of the drive system 106 with power. The power supply unit 112 preferably consists of a replaceable and rechargeable first, larger battery module 118 for the long-term power supply of the system and a second, smaller, permanently installed battery module 120 that bridges the operation of the platform for a short time when the larger battery module 118 is replaced. The first, larger battery module 118 can be attached to the drive system 106 while the drive system 106 is already mounted on the microfluidic system 102 and is in operation, as shown in Figure 11. The battery module 118, 120 can be plugged or slid on, or magnetically attached and secured by snapping into place.The battery module 118, 120 is connected to the drive system 106 via contacts for power transmission. The battery module 118, 120 and the drive system 106 are either sealed separately or the seal is created upon attachment to ensure protection against moisture. The battery module 118, 120 has a separate power connection for charging separately from the platform. Preferably, several larger battery modules 118 are used so that an empty battery module can be replaced with a charged one. This allows the battery to be replaced after a certain period of time to extend the overall cultivation time. The second, smaller battery module 120 consists of a small battery, which is always charged by the larger battery module 118 and is only discharged when the larger battery module 118 is replaced. The terms "larger" and "smaller" battery module refer to a larger or smaller energy storage capacity, respectively.

Claims

Claims 1. A cell culture platform (100) for integration into detection systems, in particular into microscopy systems, the cell culture platform (100) comprising: a microfluidic system (102); at least one cell culture component (104); and a drive system (106) comprising at least one micropump (108), a control unit (110), and a power supply unit (112).

2. Cell culture platform (100) according to claim 1, wherein the microfluidic system (102), the at least one cell culture component (104) and the drive system (106) are positively and / or non-positively coupled to the cell culture platform (100).

3. Cell culture platform (100) according to claim 1 or 2, wherein the dimensions of the cell culture platform (100) correspond to the size standard according to the ANSI / SBS 1-2004 standard for microtiter plates, in particular wherein the cell culture platform (100) has a length of 120-130 mm, a width of 80-90 mm and a height of 50 mm.

4. Cell culture platform (100) according to one of the preceding claims, wherein the microfluidic system (102) is detachably connectable to the drive system (106).

5. Cell culture platform (100) according to one of the preceding claims, wherein the at least one cell culture component (104) is part of the microfluidic system (102), in particular wherein the at least one cell culture component (104) is a cell culture chamber with an inlet and an outlet that is closed in the coupled state.

6. Cell culture platform (100) according to one of the preceding claims, wherein the at least one cell culture component (104) comprises a device, in particular a connection, for cell culture inserts.

7. Cell culture platform (100) according to one of the preceding claims, additionally comprising at least one medium reservoir (114), which is arranged in particular laterally adjacent to the at least one cell culture component (104) on the cell culture platform (100) and / or which is arranged in particular vertically adjacent to the at least one cell culture component (104).

8. Cell culture platform (100) according to one of the preceding claims, wherein the The drive system (106) is designed such that the cell culture platform can be operated for at least 24 hours, in particular at least 72 hours, further in particular at least 100 hours 9. Cell culture platform (100) according to one of the preceding claims, wherein the drive system (106) is covered by a housing (116) and / or wherein the housing (116) is connectable to the cell culture platform (100) by means of a screw connection, plug connection and / or magnetic connection and / or wherein the screw connection, plug connection and / or magnetic connection is connectable in a form-fitting manner by a seal (122) on the cell culture platform (100).

10. Cell culture platform (100) according to one of the preceding claims, wherein the at least one micropump (108) is operable by direct current.

11. Cell culture platform (100) according to one of the preceding claims, wherein the control unit (110) comprises a communication module for wireless communication between the cell culture platform (100) and a computer system.

12. Cell culture platform (100) according to one of the preceding claims, wherein the energy supply unit (112) comprises a replaceable and rechargeable first battery module (118) and a second battery module (120) fixedly connected to the cell culture platform (100).

13. Cell culture platform (100) according to claim 12, wherein the first battery module (118) has a greater storage capacity than the second battery module (120) and / or wherein the first battery module (118) can be used for the long-term supply of the cell culture platform (100) and the second battery module (120) can be used for the operation of the cell culture platform (100) when the first battery module (118) is replaced.

14. Method for execution in a detection system, in particular in a microscopy system, wherein an examination of a cell culture is carried out by means of an autonomously operable cell culture platform (100), in particular the cell culture platform (100) of claims 1-13.

15. Use of a drive system (106) on a cell culture platform (100), in particular the cell culture platform (100) from claims 1-13, wherein the drive system (106) is designed such that the cell culture platform (100) can be operated autonomously by means of the drive system (106).

16. Use of the drive system (106) of claim 15, wherein the drive system (106) comprises at least one micropump (108), a control unit (110) and a power supply unit (112).

17. Use of the drive system (106) of claim 15 or 16, wherein the Cell culture platform (100) can be operated by the drive system (106) for a running time of at least 24 hours, in particular at least 72 hours, further in particular at least 100 hours.

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