Cell culture membrane insert, and device and method for detecting at least one electrochemical variable of a cell culture

Abstract

The invention relates to a cell culture membrane insert (120) and to a device (110) and a method (210) for detecting at least one electrochemical variable of a cell culture (112). The cell culture (112) can be at least one of any cell type of at least one species in a mono- or co-culture, particularly preferable are cell cultures which are exposed to at least one gas or aerosol. The cell culture membrane insert (120) comprises - an open support (128) which is designed to expose a cell culture (112) to at least one gas or aerosol (114), wherein a membrane (130) which has at least one surface (132) is introduced into the support (128), said surface being designed to receive the cell culture (112), and the membrane (130) has o pores (134) for diffusing a nutrient medium (136) through the membrane (130) and o an electrode assembly (130) on the at least one surface (132) designed to receive the cell culture (112), said electrode assembly (130) comprising at least two electrodes, and - at least two contact points (146, 146'). Each contact point (146, 146') is designed to produce an electric connection between one of the electrodes and a respective electric contact (144, 144') which is not encompassed by the cell culture membrane insert (120). In particular, the method and the device (110) allow a time-resolved, non-invasive, and marking-free examination and monitoring of cell cultures (112) which can be exposed to at least one gas or aerosol (114) in particular, said examination and monitoring being as close to reality as possible.

Description

[0001] Cell culture membrane insert and device and method for detecting at least one electrochemical parameter of a cell culture

[0002] Field of the invention

[0003] The invention relates to a cell culture membrane insert, as well as a device and a method for detecting at least one electrochemical parameter of a cell culture. The cell culture can comprise at least one cell type of at least one species in monoculture or coculture, with cell cultures exposed to at least one gas or aerosol being particularly preferred. However, the use of other types of cell culture is possible.

[0004] State of the art

[0005] A realistic in vitro testing of cell cultures, in particular of the lungs, bronchi, mouth and throat or skin, against exposure to at least one gas or aerosol at a gas-liquid interface is recommended by the OECD as the preferred method in the field of health protection, since in submerged cultivation the gas phase is lost and particles from the aerosol are suspended in the liquid, which can influence the agglomeration status and particle surfaces, up to and including a suppression of biological effects due to the presence of proteins.

[0006] In addition to known tests on repeatedly removed samples of the nutrient medium, the investigation of the biological effect of the gas or aerosol on the cell culture, particularly in the context of molecular biological or (immuno)histological analyses, can currently only be carried out as endpoints, as this requires the removal of cell samples from the exposure chamber and the analysis usually leads to the destruction of the cell culture. Therefore, such endpoint analyses only reflect the biological state of the cell culture at the end of exposure to the gas or aerosol. This allows exactly one state to be recorded at the end of the exposure, whereas the investigation of a time-dependent effect, such as can occur during recovery phases, requires a large number of experiments and cell cultures, whereby only a relative comparison of the results is possible.

[0007] Another known method is the analysis of cell cultures that form a barrier on microporous membranes by measuring the transepithelial electrical resistance (TEER) or by determining the impedance of the cell culture. A disadvantage of this method is that it requires culturing the cell culture under submerged conditions, i.e., with a permanently covering liquid medium, preventing direct exposure of the cells to the gas or aerosol at the gas-liquid interface. Layering the cells with the liquid medium is also necessary due to the use of immersed electrodes above and below the cell culture, with the liquid medium serving as an electrically conductive electrolyte.

[0008] In this context, an online dose measurement for particle deposition on the boundary layer is also known, see e.g. WO 2008 / 116540 Al or WO 2016 / 096137 Al.

[0009] EP 2 103 933 A1 discloses a device and a method for impedance measurement at the gas-liquid interface of organotypic cell cultures on microporous membranes. For this purpose, a first electrode in the form of a submerged electrode is used, which is positioned above the membrane, while a second electrode is arranged on the floor of the culture chamber below the membrane. The first electrode is moved along the gas-liquid interface in such a way that direct contact with the cell culture is achieved for measurements. However, direct contact is only possible with a sufficiently thick cell culture layer to avoid impairment or destruction of the cell culture, and is thus limited to tissue sections; measurement on single cell layers is not feasible. Another disadvantage is that the arrangement of the second electrode at a distance from the cell culture can lead to a loss of signal.

[0010] EP 3 722 807 A1 discloses a microfluidic device for determining the transepithelial electrical resistance (TEER) of a cell layer or a cell cluster or for determining the impedance of cells, a cell layer or a cell cluster, wherein the device comprises at least one microchannel having at least one lower and one upper compartment separated by at least one porous membrane, and optionally an inner compartment, wherein the lower compartment comprises a lower wall and side walls, the upper compartment comprises an upper wall and side walls, wherein the lower wall, the upper wall, the side walls and the at least one porous membrane define volumes, and wherein at least one porous membrane comprises a surface of at least one electrode.

[0011] EP 4 119 653 A1 discloses cells on a permeable layer in a culture area provided with electrodes for measuring electrical cell properties.

[0012] Sbarana, T. et al., Dual flow bioreactor with ultrathin microporous TEER sensing membrane for evaluation of nanoparticle toxicity, Sensors and Actuators B: Chemical (2016) 223: 440- 446, show a cell resistance measuring device (TEER bioreactor) with cells on a porous substrate for testing the toxicity of nanoparticles.

[0013] Moura, JA et al., "Alternative lung cell model systems for toxicology testing strategies: Current knowledge and future outlook." Semin. Cell Dev. Biol., published electronically on January 2, 2023, 147:70-82, demonstrate lung cell models in cell culture inserts for investigating the toxicology of air pollution and inhalation of toxins. Furthermore, an air-medium contact (ALI) cell culture system with a cell culture insert is presented.

[0014] Skardal, A. et al., Multitissue interactions in an integrated three-tissue organ-on-a-chip platform. Sci. Rep. (2017) 7, 8837, pages 1–16, demonstrate a microfluidic lung cell culture device with electrodes for TEER measurement for testing toxic airborne particles or drug aerosols.

[0015] Le Prieur, E. et al., Toxicity of diesel engine exhausts in an in vitro model of lung slices in biphasic organotypic culture: Induction of a proinflammatory and apoptotic response. Arch. Toxicol. (2000) 7 4, 460-466, show an exposure chamber with a rotating culture vessel and a metal mesh for cell culture of test lung tissue.

[0016] Object of the invention

[0017] Based on this, the object of the present invention is to provide a cell culture membrane insert as well as a device and a method for detecting at least one electrochemical parameter of a cell culture, which at least partially overcome the disadvantages and limitations of the prior art mentioned.

[0018] In particular, the cell culture membrane insert, the device and the method are intended to enable a time-resolved, non-invasive and label-free realistic investigation and monitoring of cell cultures, which in particular can be exposed to at least one gas or aerosol.

[0019] Furthermore, the cell culture membrane insert, the device and the method should make it possible to obtain additional information about the general condition of the cell culture, in particular about the density or vitality of the cells contained therein.

[0020] Disclosure of the invention

[0021] This object is achieved by a cell culture membrane insert as well as a device and a method for detecting at least one electrochemical parameter of a cell culture according to the features of the independent claims. Advantageous further developments, which can be implemented individually or in any combination, are presented in the description and the dependent claims.

[0022] In a first aspect, the present invention relates to a cell culture membrane insert. The term "cell culture membrane insert" refers to a device designed to accommodate a cell culture. Within the scope of the present invention, the cell culture membrane insert serves in particular for use in an investigation of the cell culture accommodated thereby, preferably by detecting at least one electrochemical parameter of the cell culture. Furthermore, however, the cell culture membrane insert can also be used to obtain additional information about a general condition of the cell culture, in particular about a density or vitality of the cells comprised by the cell culture.

[0023] The proposed cell culture membrane insert includes:

[0024] - an open carrier which is designed to expose a cell culture to at least one gas or aerosol, wherein a membrane is introduced into the carrier which has at least one surface which is designed to receive a cell culture, wherein the membrane o has pores for the diffusion of nutrient medium through the membrane, and o an electrode arrangement on the at least one surface designed to receive the cell culture, wherein the electrode arrangement comprises at least two electrodes, and - at least two contact points, wherein each contact point is designed to establish an electrical connection between one of the electrodes and a respective electrical contact not encompassed by the cell culture membrane insert.

[0025] The term "cell culture" refers to a cell, a cell layer or a cell cluster that comprises at least one cell type of at least one species in mono- or co-culture. Cultures of eukaryotic cells are particularly preferred. Cell cultures that are particularly preferred within the scope of the present invention can be exposed to at least one gas or aerosol, in particular cell cultures of vertebrates, in particular of mammals, for example human cell cultures, cell cultures of the mouse (Mus musculus) or rat (Rattus norvegicus), furthermore particularly preferred are cell cultures of the lungs, the bronchi, the mouth and throat or the skin. The cell culture used within the scope of the present invention can be primary cells, a section through a cell tissue, cells grown on a membrane or the culture of a known cell line, such ashuman lung cell lines A549, BEAS-2B, HBE, in monocultures or in co-culture systems of two or more cell types, e.g., one of the aforementioned epithelia in combination with THP-1-induced macrophages. However, the use of a different cell culture is possible. In particular, the cell culture may be suitable as a biosensor for at least one gas or aerosol that is classified or suspected of being an air pollutant for the cell culture.

[0026] The term "gas" refers to a substance or a mixture of at least two substances, where the substance comprises atomic or molecular particles that move freely apart from one another and fill an available space. In contrast, the term "aerosol" refers to a mixture of at least one gas and particles or fibers. The term "particles" refers to particles that can be liquid or solid, whereby these have a so-called "aspect ratio", i.e. a ratio of the length to the diameter of the particle, of at most 3:1; whereas the term "fibers" refers to particles that have an aspect ratio higher than 3:1.

[0027] The cell culture membrane insert proposed herein comprises an open support configured to expose a cell culture to at least one gas or aerosol. The term "support" refers to a mechanically stable device configured to temporarily or permanently accommodate at least one element. The term "open" refers to external accessibility of an internal volume of an element, in this case the support, in particular in order to apply the at least one cell culture to the membrane or to allow it to grow there and to expose it to the gas or aerosol. For this purpose, the proposed cell culture membrane insert can have at least an inner side and an outer edge. For this purpose, the cell culture membrane insert can preferably be designed in the form of a cup with an inner side and an outer edge; however, another design form is conceivable.

[0028] A membrane is introduced into the carrier, preferably into the inner side of the carrier, of the cell culture membrane insert. The term "membrane" here refers to a flat, extended semipermeable element, whereby the expression "flat, extended" expresses that the membrane has a membrane body with a diameter and a layer thickness that is preferably constant across the diameter. A biocompatible plastic is preferably suitable as a material for the membranes, whereby the plastic can preferably be selected from polycarbonate, polyethylene terephthalate, polytetrafluoroethylene, polystyrene, cellulose or cellulose mixed esters; however, the use of another plastic or a combination of at least two plastics is possible. In particular, to improve cell adhesion, at least one additional treatment of the membrane can be carried out or at least one coating can be applied.The diameter of the membrane can exceed the layer thickness of the membrane by at least a factor of 10, preferably by a factor of 100, particularly preferably by at least a factor of 1000. The membrane can preferably have a layer thickness of 2 μm to 100 μm, particularly preferably from 2 μm to 50 μm, in particular from 5 μm to 20 μm, for example from 10 μm. The membrane can preferably have a diameter of 2 mm to 100 mm, particularly preferably from 5 mm to 50 mm, in particular from 10 mm to 30 mm. A particularly preferred diameter for the membrane can be a standard size, preferably selected from 6 mm, 12 mm, 24 mm, 14" (6.35 mm), 14" (12.7 mm), or 1" (25.4 mm), in particular to enable the use of standard formats and compatibility with other devices, especially for further studies on the same cell culture. However, other values ​​for layer thickness and membrane diameter are possible.

[0029] The membrane has at least one surface designed to accommodate a cell culture. The term “surface” here refers to an area of ​​the membrane that is arranged perpendicular to the layer thickness along the diameter of the membrane. The at least one surface of the membrane can be uncoated or have a coating. The membrane can particularly preferably be designed as a disc-shaped element that comprises two opposing surfaces; however, another design of the membrane is conceivable. The term “accommodation” refers to the ability of at least one of the surfaces of the membrane to provide a support that is suitable for the growth and adherent cultivation of at least one type of cell culture.In a preferred embodiment, one surface of the membrane can serve to completely accommodate the cell culture, while at least one other surface of the membrane can remain free of the cell culture. In an alternative embodiment, at least two surfaces of the membrane can be configured to accommodate the cell culture. For example, the membrane can be configured such that epithelia can be applied to a first surface of the membrane, while an endothelium or fibroblasts can form on a second surface of the membrane, which is preferably arranged opposite the first surface of the membrane. Further embodiments of the membrane are conceivable.

[0030] This distinguishes the present invention in particular from the disclosure in EP 3 722 807 A1, according to which the membrane and the associated support have microchannels with microfluidic dimensions of at most 1.5 mm. Due to the narrow microchannels, the device of EP 3 722 807 A1 can only determine the influence of at least one gas on the cell culture, since exposure to at least one aerosol containing liquid and / or solid particles can easily lead to blockage of the microchannels. In addition, a linear deposition of particles occurs in the microchannels over successively flowed over areas, thus generating a gradient which prevents the simultaneous exposure of different areas in a cell culture to the aerosol and thus a determination of the effect on the cell culture with the same local dose of the aerosol.On the one hand, the gradient in the microchannels forms due to the changing particle concentration in the aerosol phase, and on the other hand, because the rate of aerosol deposition depends on the size of the particles it contains. Compared to EP 3 722 807 A1, the present invention is thus suitable for the simultaneous exposure of different areas of a cell culture to a gas or aerosol and thus for determining the effect on the cell culture at the same local dose within the cell culture.

[0031] According to the above definition, the "membrane" refers to a semipermeable element, where the term "semipermeable" expresses that the membrane is designed such that the membrane can be traversed by at least one first substance acting on the membrane, while the membrane can retain at least one other substance acting on the membrane. In the case of the present invention, the membrane has pores which are designed for the diffusion of nutrient medium through the membrane. Here, the term "pores" refers to a plurality of openings which penetrate the membrane body, preferably perpendicular to the surface. In this way, a cell culture which forms on at least one of the surfaces of the membrane which is designed to accommodate the cell culture can be supplied with nutrient medium.The term "nutrient medium" refers to a medium that is itself designed to supply the cell culture and / or that contains a substance designed to supply the cell culture. In particular, the term refers to media that are suitable for the growth and / or cultivation of the cell cultures or cell lines mentioned here. Furthermore, the nutrient medium can serve, in particular, to promote cell metabolism and to balance osmotic pressure. A particularly suitable nutrient medium can be RPMI-1640 medium, Minimum Essential Medium, Hank's Medium, or Dulbecco's Modified Eagle's Medium, both with and without additives such as serum, especially fetal bovine serum. Buffer substances such as HEPES (2-(4-(2-hydroxyethyl)-l-piperazinyl)ethanesulfonic acid) can be used as further alternatives to nutrient medium.

[0032] According to the present invention, the membrane further comprises an electrode arrangement attached to the at least one surface of the membrane designed to accommodate the cell culture. The term "electrode arrangement" refers here to a configuration of electrically conductive elements referred to as "electrodes", wherein the configuration is designed to provide an electrical voltage and / or an electrical current in order to thereby be able to measure an electrical current and / or an electrical voltage. At least one geometric parameter of the electrode arrangement, in particular the size, spacing, layer thickness and shape of the included electrodes, can be selected such that sensitive detection and / or changes in the state of the cells in the cell culture are possible despite an electrically insulating gas-liquid interface directly above the cell culture.

[0033] A suitable material for the electrode array is a metal, graphite, or conductive polymer, with the metal preferably being selected from gold, platinum, titanium, or iridium. The use of another biocompatible metal or a biocompatible alloy of at least two metals or conductive metal oxides or metal nitrides is also possible. The material for the electrode array can be applied to the membrane surface designed to receive the cell culture using a known coating process, in particular by vapor deposition, sputtering, or printing of the material. At least two electrodes are required to perform such a measurement.As explained in more detail below, the electrode arrangement is designed to detect at least one electrochemical quantity, in particular to determine an impedance or a transepithelial electrical resistance, of the cell culture.

[0034] In a particularly preferred embodiment, the electrode arrangement can be configured for spatially resolved detection of at least one electrochemical parameter of the cell culture when an electrical voltage and / or an electrical current is applied to the at least two electrodes. The term "spatially resolved" refers to the assignment of a recorded value for an electrical voltage and / or an electrical current to a location where the recorded value for the electrical voltage and / or the electrical current is present.

[0035] Furthermore, the cell culture membrane insert comprises at least two contact points. Each of the at least two contact points is designed to establish an electrical connection between an electrode on the at least one surface of the membrane of the cell culture membrane insert and an electrical contact that is not encompassed by the cell culture membrane insert. The term “contact point” refers here to a device that is designed to directly or indirectly establish contact with an external device for detecting at least one electrochemical parameter of a cell culture. In this way, external control and / or regulation of the electrode arrangement located in the cell culture membrane insert can be carried out in order to be able to carry out the desired measurement of an electrical current and / or an electrical voltage in order to thereby detect the at least one electrochemical parameter of the cell culture.

[0036] In a particular embodiment, the at least two contact points comprised by the cell culture membrane insert can be designed as inductive contacts that can establish contact with the device for detecting at least one electrochemical variable of a cell culture through contactless transmission by means of induction. Alternatively or additionally, the at least two contact points can be designed as electrically conductive contacts that can establish contact with the device for detecting the at least one electrochemical variable of the cell culture by means of direct contact. The term “direct contact” here refers to a direct electrically conductive connection that can be established in particular by means of clamping, plugging or coupling. In a particular embodiment, the at least two contact points can be attached directly to the electrode arrangement located on the membrane of the cell culture membrane insert.In a particularly preferred embodiment, the at least two contact points can each be connected to the electrodes located on the membrane by means of a conductor arrangement. The term "conductor arrangement" refers to an arrangement of an electrical conductor designed to establish an electrically conductive connection between two points. The same material as the electrode arrangement can be suitable for the conductor arrangement, in particular a metal or graphite.In this embodiment, the inner side of the carrier can preferably comprise at least two electrical conductor arrangements, each of which is designed to establish a continuous electrical connection between one of the electrodes and the respective electrical contact on the device not encompassed by the cell culture membrane insert, by establishing an electrical connection from each electrode in the electrode arrangement via the respective associated electrical conductor arrangement and the respective associated contact point to the respective associated contact on the device. In this case, the at least two contact points can preferably be attached to the outer edge of the carrier, in particular in such a way that an electrical connection can be formed in each case to a respective associated contact, which can be mounted in at least one cover of the device, described in more detail below.However, further designs of the at least two contact points included in the cell culture membrane insert are conceivable.

[0037] In a further aspect, the present invention relates to a device for detecting at least one electrochemical parameter of a cell culture exposed to at least one gas or aerosol. The proposed device comprises:

[0038] - an open housing configured to receive o at least one cell culture membrane insert disclosed herein, o at least one nutrient medium for the cell culture, and o at least one supply for the at least one gas or aerosol;

[0039] - at least two contacts, each of which is designed to establish an electrical connection to the at least two contact points comprised by the cell culture membrane insert; and

[0040] - at least one connection to a measuring and control device.

[0041] The device can be designed in one piece or in multiple parts, with the one-piece design comprising the housing, the at least two contacts, and the at least one connection to the measuring and control device being preferred. Furthermore, the present device can comprise at least one further element, which may or may not be mentioned herein. As already mentioned above, the term "open" refers to the accessibility of an internal volume of an element, here the housing, from the outside, in particular in order to introduce the at least one cell culture membrane insert and the at least one nutrient medium for the at least one cell culture and to supply the at least one gas or aerosol for acting on the at least one cell culture. Other types of design of the device are conceivable.

[0042] As mentioned above, the device comprises a housing. The term “housing” refers to a mechanical device that is designed to receive and store at least one further element. In the case of the present invention, this is the at least one cell culture membrane insert, the at least one nutrient medium for the at least one cell culture, the at least one supply for the at least one gas or aerosol and, preferably, at least one discharge for the at least one gas or aerosol. In a preferred embodiment, the housing can

[0043] - for simultaneously accommodating a plurality of cell culture membrane inserts;

[0044] - to supply the cell cultures in the majority of the cell culture membrane inserts with the same or a different nutrient medium; and

[0045] - be configured to expose the cell cultures in the majority of the cell culture membrane inserts to the same or a different gas or aerosol. When exposing the cell cultures to the same gas or aerosol, a change in the concentration of the gas or aerosol can occur, preferably in the form of a dilution series. The majority of cell culture membrane inserts can, in particular, comprise 2, 3, 4, 6, 12, 24, 48, or 96 cell culture membrane inserts. However, other housing designs are possible.

[0046] To accommodate the at least one cell culture membrane insert, the housing can preferably have at least one holding element configured for this purpose. In a particular embodiment, the at least one holding element can be designed in the form of a uniform, preferably annular holder; however, other configurations of the at least one holding element are possible. For example, to accommodate 2, 3, 4, 6, 12, 24, 48, or 96 cell culture membrane inserts, a correspondingly designed multi-well plate can be used. In a particularly preferred embodiment, the at least one cell culture membrane insert can be detachably attached to the at least one holding element.Advantageously, the at least one cell culture membrane insert can thus be interchangeable—particularly in contrast to the disclosure in EP 3 722 807 A1—to enable consecutive use of the device for multiple identical or different cell cultures, each of which is housed in a different cell culture membrane insert. This difference from the disclosure in EP 3 722 807 A1 can, in particular, also allow flexible installation and removal of the membrane, thus enabling sequential addition or removal of different cell types at different times after the respective cell attachment.Further advantages of this difference include the possibility of using standard formats and compatibility with other devices, particularly for further studies on the same cell culture, thus enabling industrial suitability and / or the investigation of additional cell parameters in standard systems. Furthermore, it is advantageous if only the cell culture membrane insert is designed as a disposable product, while the device can be used over a long period of time. Further advantages are conceivable.

[0047] To supply the cell cultures in the at least one cell culture membrane insert with at least one nutrient medium, at least a portion of the housing, preferably at least up to the height of the at least one holding element for receiving the at least one cell culture membrane insert, can be designed as a vessel configured to receive and store a liquid, in this case the at least one nutrient medium. Alternatively or additionally, the housing can have at least one device for supplying and / or discharging the at least one nutrient medium.

[0048] In order to supply the at least one cell culture with the at least one gas or aerosol, the housing comprises at least one supply for the at least one gas or aerosol. In a particular embodiment, the housing can have at least one opening for this purpose, preferably above the height of the at least one holding element for receiving the at least one cell culture membrane insert, which opening is designed at least for supplying the at least one gas or aerosol. Alternatively or additionally, the device can comprise at least one cover. The term “cover” here refers to an element that is designed to close the at least one opening of the housing above the height of the at least one holding element for receiving the at least one cell culture membrane insert.The cover can preferably be attached to the housing in a detachable or foldable manner to ensure accessibility to the interior volume of the housing from the outside, in particular to enable insertion of the at least one cell culture membrane insert into the housing. In a particularly preferred embodiment, the at least one cover can have at least one opening configured at least for supplying the at least one gas or aerosol, as well as at least one further opening configured at least for discharging the at least one gas or aerosol. However, other embodiments of the at least one cover are possible.

[0049] As further mentioned above, the device comprises at least two contacts, which are ultimately configured to establish an electrical connection to the at least two electrodes comprised by the electrode arrangement of the cell culture membrane insert. In a preferred embodiment, the at least two contacts can be attached to the housing. Alternatively or additionally, the at least two contacts can be attached to the at least one cover. In a particular embodiment, the at least two contacts can be designed as inductive contacts, which can establish contact with the at least two contact points comprised by the cell culture membrane insert through contactless transmission by means of induction.Alternatively or additionally, the at least two contacts can be designed as electrically conductive contacts that can establish contact with the at least two contact points encompassed by the cell culture membrane insert by means of direct contact. As already mentioned above, the term "direct contact" refers to a direct electrically conductive connection, which can be established in particular by means of clamping, plugging, or coupling.

[0050] In a preferred embodiment, the device can be designed to accommodate the cell culture membrane insert in the device as positively as possible. Particularly preferably, both the cell culture membrane insert, in particular the membrane and electrode arrangement comprised thereby, and the device, in particular the at least one holding element, the at least one inlet, and the at least one outlet for the at least one gas or aerosol, can be designed to be rotationally symmetrical. However, other embodiments of both the device and the cell culture membrane insert intended for accommodation by the device are possible.

[0051] The present device is designed to detect at least one electrochemical parameter of a cell culture, in particular while at least one surface of the cell culture facing away from the membrane is at least temporarily exposed to the at least one gas or aerosol and remains free of the nutrient medium during this time. The term “electrochemical parameter” refers to a property of an object that can be measured using devices and methods known from electrochemistry, the object here being at least one cell culture that is applied to at least one surface of a membrane encompassed by the cell culture membrane insert. As explained in more detail above, the detection of the at least one electrochemical parameter takes place by means of an electrode arrangement that is attached to the at least one surface of the membrane designed to accommodate the cell culture.In this way, a time-resolved, non-invasive and label-free realistic investigation and monitoring of cell cultures, which may in particular be exposed to at least one gas or aerosol, is enabled.

[0052] In a particularly preferred embodiment, the electrochemical quantity comprises an impedance, which is determined by means of the electrode arrangement across the cell culture or, preferably, across a region of the cell culture. The term "impedance" refers to a complex electrical resistance in an alternating current circuit, which is provided by the electrode arrangement. Alternatively or additionally, the electrochemical quantity can be a so-called "transepithelial electrical resistance", or "TEER" for short, which indicates the electrical resistance through a cell culture. The present invention enables, in particular, the detection of the impedance of the cell cultures adherently cultivated on the membrane, preferably over a range of at least 1 Hz, at least 5 Hz, at least 10 Hz, up to a maximum of 100 MHz, a maximum of 500 MHz, and a maximum of 1000 MHz.While the determination of transepithelial electrical resistance, which is preferably used to determine a cell barrier, usually requires a sufficiently dense cell layer or a cell barrier, this is not necessary for the measurement of impedance.

[0053] In a particular embodiment, the device can further comprise at least one immersion electrode. The terms "immersion electrode" or "immersion electrode" refer here to an electrically conductive element referred to as an "electrode," which is designed to be inserted, in particular spaced apart from a wall, into a liquid, preferably an electrically or ionically conductive electrolyte. Within the scope of the present invention, the at least one immersion electrode can be designed both for insertion into the at least one nutrient medium for the cell culture and for detecting at least one further electrochemical parameter of the cell culture.The at least one further electrochemical parameter of the cell culture may preferably be a density or vitality of the cells comprised in the cell culture or a determination of a redox tracer or a chamber potential; however, the detection of another further electrochemical parameter of the cell culture is conceivable.

[0054] For further details regarding the present device, reference is made to the description of the cell culture membrane insert. In a further aspect, the present invention relates to a method for detecting at least one electrochemical parameter of a cell culture exposed to at least one gas or aerosol. The present method comprises at least the following method steps, which can be performed sequentially in the specified order or, preferably, at least partially simultaneously.The method steps are at least: a) providing at least one cell culture membrane insert disclosed herein; b) applying the cell culture to at least one surface of a membrane of the cell culture membrane insert; c) providing at least one nutrient medium for the cell culture; d) exposing the cell culture to at least one gas or aerosol; e) detecting at least one electrochemical parameter of the cell culture by means of at least one device disclosed herein.

[0055] According to step a), at least one cell culture membrane insert, described in more detail above and below, is provided, which is designed to accommodate a cell culture. The cell culture membrane insert comprises a membrane, wherein, according to step b), the cell culture is applied to at least one surface of the membrane. According to step c), at least one nutrient medium for the cell culture is further provided.

[0056] According to step e), at least one electrochemical parameter of the cell culture is detected by means of at least one device described in more detail above and below for detecting the at least one electrochemical parameter of the cell culture, which is exposed to at least one gas or aerosol according to step d). In a particularly preferred embodiment, at least one surface of the cell culture facing away from the membrane is at least temporarily exposed to the at least one gas or aerosol during the detection of the at least one electrochemical parameter of the cell culture according to step e), and remains free of the nutrient medium during this time.In this embodiment, the at least one surface of the cell culture facing away from the membrane can also be covered with nutrient medium, at least outside of the detection of the at least one electrochemical variable of the cell culture according to step e), in particular to enable the cultivation of cells that are particularly sensitive to at least one gas or aerosol. Alternatively or additionally, the cell culture membrane insert can be removed from the device, in particular during a period between two consecutive measurements, and stored in another vessel, preferably filled with nutrient medium. The "cultivation of the cell culture" refers in particular to the cultivation of cells or cell cultures in a suitable nutrient medium under conditions that allow the growth of the respective cells or cell cultures.Cultivation can be understood in particular as the cultivation of cell cultures at a suitable air humidity, a suitable CO2 content and a suitable temperature. Cultivation preferably takes place at an air humidity of more than 90% relative humidity, more preferably at approximately 95% relative humidity, a CO2 content of 0 to 10 vol.%, preferably approximately 0 or 5 vol.%, and a temperature of at least 20 °C to a maximum of 45 °C; cultivation at a temperature of approximately 37 °C is particularly suitable. In addition, other substances, in particular active ingredients for manipulating the cell culture, can be added to the nutrient medium in order to be able to influence the behavior of the cell culture after exposure to particles, such as viruses.

[0057] For further details regarding the present method, reference is made to the description of the cell culture membrane insert and the device according to the invention.

[0058] In a further aspect, the present invention relates to the use of a device for detecting at least one electrochemical parameter of a cell culture exposed to at least one gas or aerosol. The cell culture may comprise at least one cell type of at least one species in monoculture or coculture, with cell cultures exposed to at least one gas or aerosol being particularly preferred. However, the use of a different type of cell culture is possible.

[0059] In a further aspect, the present invention relates to a use of a cell culture membrane insert for detecting at least one electrochemical variable of a cell culture exposed to at least one gas or aerosol.

[0060] For further details regarding the present uses, reference is made to the description of the cell culture membrane insert, the device according to the invention and the method according to the invention.

[0061] Herein, the terms "have," "have," "comprise," or "include," or any grammatical variations thereof, are used non-exclusively. Accordingly, these terms can refer both to situations in which, besides the features introduced by these terms, no further features are present, or to situations in which one or more further features are present. For example, the expression "A has B," "A has B," "A comprises B," or "A includes B" can refer both to the situation in which, apart from B, no further element is present in A (i.e., a situation in which A consists exclusively of B), and to the situation in which, in addition to B, one or more further elements are present in A, for example, element C, elements C and D, or even further elements.

[0062] Furthermore, it should be noted that the terms "at least one" and "one or more," as well as grammatical variations of these terms, when used in connection with one or more elements or features and intended to express that the element or feature may be provided singly or multiple times, are generally used only once, for example when the feature or element is first introduced. When the feature or element is subsequently mentioned again, the corresponding term "at least one" or "one or more" is generally no longer used, without this limiting the possibility that the feature or element may be provided singly or multiple times.

[0063] Furthermore, the terms "preferably", "in particular", "for example" or similar terms are used herein in connection with optional features, without limiting alternative embodiments. Thus, features introduced by these terms are optional features, and these features are not intended to limit the scope of the claims and in particular the independent claims. Thus, as those skilled in the art will recognize, the invention can also be carried out using other embodiments. Similarly, features introduced by "in one embodiment of the invention" or by "in an embodiment of the invention" are understood to be optional features, without limiting alternative embodiments or the scope of the independent claims.Furthermore, these introductory expressions are intended to leave untouched all possibilities of combining the features introduced thereby with other features, be they optional or non-optional.

[0064] Brief Description of the Figures Further details and features of the present invention will become apparent from the following description of preferred embodiments, particularly in conjunction with the dependent claims. The respective features can be implemented individually or in combination with one another. However, the invention is not limited to the embodiments. The embodiments are schematically illustrated in the following figures. Like reference numerals in the figures denote like or functionally identical elements, or elements that correspond to one another in terms of their functions. In detail:

[0065] Figure 1 is a schematic representation of a preferred embodiment of a device according to the invention for detecting at least one electrochemical variable of a cell culture exposed to at least one gas or aerosol;

[0066] Figure 2 is a schematic representation of a further preferred embodiment of the device according to the invention;

[0067] Figure 3 shows schematic representations of three preferred embodiments of an electrode arrangement; and

[0068] Figure 4 is a schematic representation of a preferred embodiment of a method according to the invention for detecting at least one electrochemical variable of a cell culture exposed to at least one gas or aerosol.

[0069] Description of the embodiments

[0070] Figure 1 shows a schematic cross-sectional representation of a preferred embodiment of a device 110 according to the invention for detecting an electrochemical parameter of a cell culture 112, which in this case is exposed to an aerosol 114. The aerosol 114 shown as an example in Figure 1 comprises a gas in which gas-borne particles 116 and fibers 118 are located; however, the use of a gas without particles or a different type of aerosol 114 is possible. The gas can be, for example, air, in which the gas-borne particles 116 and the fibers 118 are considered to be air pollutants that are classified or suspected of being harmful to the cell culture 112. Alternatively, the particles 116 can also transport active substances or be of biological origin, such as viruses or bacteria.

[0071] Figure 1 further shows a cell culture membrane insert 120, which is inserted into an open housing 122 encompassed by the device 110. The housing 122 is designed to accommodate the cell culture membrane insert 120, in particular by having a sufficiently large opening 124 and retaining elements 126, 126', each in the form of a support, which can releasably receive the cell culture membrane insert 120. Thus, the cell culture membrane insert 120 can advantageously be exchangeable, thus enabling the device 110 to be used successively for several identical or different cell cultures 112, each of which is housed in a different cell culture membrane insert 120.In particular, the cell culture membrane insert 120 can also allow flexible installation and removal of the membrane 130, so as to enable sequential addition or removal of different cell types at different times after the cells have attached to the membrane 130.

[0072] The cell culture membrane insert 120 shown as an example in Figure 1 comprises an open carrier 128 which is designed to expose the cell culture 112 to the aerosol 114. A membrane 130 is introduced into the carrier 128 of the cell culture membrane insert 120, which membrane has at least one surface 132 designed to receive the cell culture 112, which is applied here, for example, in the form of a monocellular layer on the surface 132 of the membrane 130. The membrane 130 can preferably have a layer thickness of 2 μm to 100 μm, particularly preferably of 2 μm to 50 μm, in particular of 5 μm to 20 μm, for example of 10 μm, and a diameter of 2 mm to 100 mm, particularly preferably of 5 mm to 50 mm, in particular of 10 mm to 30 mm. As mentioned above, a particularly preferred diameter for the membrane 130 may be selected from 6 mm, 12 mm, 24 mm (6.35 mm), 'A' (12.7 mm), or 1" (25.4 mm).However, other values ​​for layer thickness and diameter of the membrane 130 are possible.

[0073] As mentioned above, the cell culture 112 can in particular be a cell, a cell layer, or a cell cluster comprising at least one arbitrary cell type of at least one species in monoculture or coculture. Particularly preferred cell cultures 112 can be exposed to at least one gas or aerosol, in particular cell cultures of the lungs, bronchi, mouth and throat, or skin. The cell culture 112 can preferably be primary cells, a section through a cell tissue, cells grown on a membrane, or the culture of a known cell line. In particular, the cell culture 112 can be suitable as a biosensor for the aerosol 114.

[0074] The membrane 130 is preferably a biocompatible plastic membrane into which a plurality of pores 134 are introduced, preferably with a constant pore density across the membrane 130. The pores 134 are configured for the diffusion of nutrient medium 136 through the membrane 130 to the cell culture 112. The nutrient medium can represent at least one substance that is itself suitable for supplying the cell culture 112 or, alternatively or additionally, can comprise at least one substance that is suitable for supplying the cell culture 112. In particular, for a better illustration of the membrane 130 and the pores 134 located therein, reference is made to the enlarged section 138 in Figure 1.

[0075] As Figure 1 schematically shows, the housing 122 can be configured to accommodate a single cell culture membrane insert 120. In an alternative embodiment (not shown), however, the housing 122 can be configured to accommodate a plurality, for example, 2, 3, 4, 6, 12, 24, 48, or 96, cell culture membrane inserts 120. In this case, the cell cultures 112 in the plurality of cell culture membrane inserts 112 can be supplied with the same and / or a different nutrient medium 136. Likewise, the cell cultures 112 in the plurality of cell culture membrane inserts 120 can be exposed to the same and / or a different aerosol 114. However, other embodiments of the cell culture membrane insert 120 and / or the housing 122 of the device 110 configured to accommodate the cell culture membrane insert 120 are conceivable.

[0076] According to the invention, the membrane 130 has an electrode arrangement 140 on the at least one surface 132 configured to receive the cell culture. The electrode arrangement 140 denotes a configuration of at least two electrodes configured to provide an electrical voltage and / or an electrical current in order to thereby enable a measurement of an electrical current and / or an electrical voltage. A particularly suitable material for the electrodes is metal, preferably gold, or graphite, although the use of another biocompatible metal or a biocompatible alloy is also possible. The electrode arrangement 140 schematically illustrated in Figure 1 is configured to detect at least one electrochemical variable, in particular to determine an impedance or a transepithelial electrical resistance (TEER), of the cell culture 112.The determination of the TEER can preferably be used to determine a cell barrier. The determination of the impedance in a range of at least 1 Hz, at least 5 Hz, at least 10 Hz, up to a maximum of 100 MHz, at most 500 MHz, at most 1000 MHz, can be used in particular to determine information about a metabolism of the cell culture 112; however, other uses are conceivable. The electrode arrangement 140 can in particular be designed such that it enables a spatially resolved detection of the at least one electrochemical variable of the cell culture 112. As schematically illustrated in Figure 3, the electrode arrangement 140 can for this purpose be designed in particular in the form of electrode fingers or spirals running parallel to one another; however, further embodiments of the electrode arrangement 140 are conceivable.

[0077] The housing 122 of the device 110 is further configured to accommodate the nutrient medium 136 for the cell culture 112. As schematically illustrated in Figure 1, the nutrient medium 136 can be introduced into the housing 122 through the opening 124. Alternatively or additionally (not illustrated), the housing 122 can have a supply line and / or outlet for the nutrient medium 136. The height of the nutrient medium 136 in the housing 122 can in principle be adjusted as desired; however, it is particularly advantageous if at least one surface of the cell culture 112 facing away from the membrane 130 remains at least temporarily free of the nutrient medium 136, while the detection of the at least one electrochemical parameter of the cell culture 112 is carried out by the aerosol 114.At least outside of the detection of the at least one electrochemical parameter of the cell culture 112, the surface of the cell culture 112 facing away from the membrane 130 can also be covered with the nutrient medium 136, in particular to enable better cultivation of the cell culture 112. In an alternative embodiment, the cell culture membrane insert 120 can be removed from the device 110, in particular between two measurements, in particular to be stored in another vessel, preferably filled with nutrient medium, and / or to be subjected to at least one further examination.

[0078] The housing 122 of the device 110 is further configured such that it has at least one inlet 142 for the aerosol 114. In the exemplary embodiment according to Figure 1, the opening 124 of the housing 122 can serve this purpose; however, further embodiments are possible; see in particular the exemplary embodiment according to Figure 2. In this way, in particular, exposure of the cell culture 112 located in the cell culture membrane insert 120 to the gas or aerosol 114 is enabled. In addition to sedimentation of an aerosol cloud as a collective on the cell culture 112, the present cell culture membrane insert 120 can also enable diffusion and / or electrostatic interaction of the particles 116 from the aerosol 114 with the cell culture 112. In a particularly preferred embodiment, the cell culture membrane insert 120, the membrane 130, the electrode arrangement 140 and the feed 142 for the aerosol 114 can be designed rotationally symmetrically.In this embodiment, the holding elements 126, 126' can preferably be designed as a uniform, annular holder. The exemplary embodiment of the device 110 shown schematically in Figure 1 further comprises two contacts 144, 144', each of which is configured to establish an electrical connection to the electrodes comprised by the electrode arrangement 140 of the cell culture membrane insert 120. To establish this electrical connection, the cell culture membrane insert 120 shown in Figure 1 has two contact points 146, 146'. In the exemplary embodiment according to Figure 1, the contact points 146, 146' comprised by the cell culture membrane insert 120 are designed as inductive contacts, which enable contactless transmission by means of induction to the contacts 144, 144' comprised by the device 110.Alternatively or additionally, further embodiments are possible; see in particular the embodiment according to Figure 2, in which the contact points 146, 146' are designed as electrically conductive contacts that can establish the electrical connection to the contacts 144, 144' comprised by the device 110 by means of direct contact.

[0079] The exemplary embodiment of the device 110 schematically illustrated in Figure 1 further comprises a connection 148 to a measuring and control device 150. The measuring and control device 150 itself can be controlled by means of an electronic communications device, preferably as a smartphone 152, tablet, or laptop. Alternatively or additionally (not illustrated), an embodiment of the measuring and control device 150 as an electronic communications device, preferably as a smartphone 152, tablet, or laptop, is also possible. The electronic communications device, preferably the smartphone 152, tablet, or laptop, can have a screen 154 on which a diagram 156 schematically illustrates a profile of the at least one electrochemical variable, in particular the impedance or the transepithelial electrical resistance, of the cell culture 112. Other embodiments are, however, possible.

[0080] Figure 2 shows a schematic cross-sectional view of another, likewise preferred embodiment of the device 110 according to the invention for detecting an electrochemical parameter of the cell culture 112 exposed to the aerosol 114. The aerosol 114 shown as an example in Figure 2 also comprises a gas in which the gas-borne particles 116 and the fibers 118 are located.

[0081] The exemplary device 110 according to Figure 2 differs from the exemplary device 110 according to Figure 1 in that the volume formed by the housing 122 is provided with a cover 160. As schematically shown in Figure 2, the cover 160 has the inlet 142 for the aerosol 114 and an outlet 162 for the aerosol 114. In this way, a controlled exposure of the cell culture 112 located in the cell culture membrane insert 120 is enabled, in particular when the cell culture 112 is exposed to a directed laminar flow 164. This can be supported in particular by the fact that the cell culture membrane insert 120, the membrane 130, the electrode arrangement 140, the inlet 142 and the outlet 162 for the aerosol 114 can be designed to be rotationally symmetrical.

[0082] The exemplary device 110 according to Figure 2 further differs from the exemplary device 110 according to Figure 1 in that the contacts 144, 144', which are configured to establish the electrical connection with the contact points 146, 146' of the cell culture membrane insert 120, are attached to the cover 160. The electrical connection between the contact points 146, 146' of the cell culture membrane insert 120 and the contacts 144, 144' of the device 110 can be established, in particular, by means of clamping, plugging, or coupling; however, other types of contacting are possible.

[0083] To establish an electrical connection between each of the electrodes in the electrode arrangement 140 and the contact points 146, 146' of the cell culture membrane insert 120 with the contacts 144, 144' attached to the cover 160, a respective electrical conductor arrangement 166, 166' is further applied to the inside of the carrier 128. In this way, a continuous electrical connection can be established from each electrode in the electrode arrangement 140 via the respective electrical conductor arrangement 166, 166' and the respective contact point 146, 146 to the respective contact 144, 144' on the device 110.

[0084] The exemplary device 110 according to Figure 2 further differs from the exemplary device 110 according to Figure 1 in that it has a submerged electrode 168 that is introduced into the nutrient medium 136. The submerged electrode 168 can be configured, in particular, to detect at least one further electrochemical parameter of the cell culture 112, preferably a density or vitality of the cells comprised by the cell culture 112, or to determine a redox tracer or a chamber potential; however, the detection of another further electrochemical parameter of the cell culture 112 is conceivable.

[0085] For further details regarding the device 110 according to Figure 2, reference can be made to the description of the device 110 according to Figure 1. Furthermore, it should be noted that further embodiments of the device 110 are conceivable, in particular any combination of features from the exemplary embodiments according to Figures 1 and 2.

[0086] Figure 4 shows a schematic representation of a preferred embodiment of a method 210 according to the invention for detecting the at least one electrochemical variable of the cell culture 110 exposed to the aerosol 114, which comprises, for example, the gas-borne particles 116 and the fibers 118.

[0087] In a provision step 212 according to step a), the at least one cell culture membrane insert 120, which is configured to receive the cell culture 112, is provided. The cell culture membrane insert 112 comprises the membrane 130, wherein in an application step 214 according to step b), the cell culture 112 is applied to the at least one surface 132 of the membrane 130. In a further provision step 216 according to step c), the nutrient medium 236 for the cell culture 112 is provided.

[0088] In an exposure step 218 according to step d), the cell culture 112 is exposed to the aerosol 114. In this case, the exposure of the cell culture 112 located in the cell culture membrane insert 120 can take place in particular according to the embodiment according to Figure 1 by means of an aerosol cloud that sediments as a collective onto the cell culture 112, or according to the embodiment according to Figure 2 by means of a directed laminar flow 164.

[0089] In a detection step 220 according to step e), the at least one electrochemical variable of the cell culture 112 is detected by means of the device 110 for detecting the at least one electrochemical variable of the cell culture 112, which is exposed to the aerosol 114 according to the exposure step 218. It is particularly preferred if the at least one surface of the cell culture 112 facing away from the membrane is at least temporarily exposed to the aerosol 114 during the detection of the at least one electrochemical variable of the cell culture 112 in the detection step 220, and remains free of the nutrient medium 136 during this time.

[0090] In this case, the detection of at least one electrochemical variable of the cell culture 112 during the detection step 220 can be carried out, in particular for monitoring the cell cultures 112 over a longer period of time, during which the type, composition, and / or concentration of the aerosol 114 can change during the exposure step 218. For further details regarding the present method 210, reference is made to the description of the device according to the invention in the embodiments according to Figures 1 and 2. List of Reference Symbols

[0091] 110 Device for detecting an electrochemical quantity of a cell culture

[0092] 112 Cell culture

[0093] 114 Aerosol

[0094] 116 gas-borne particles

[0095] 118 gas-borne fibers

[0096] 120 cell culture membrane insert

[0097] 122 open case

[0098] 124 Opening

[0099] 126, 126' holding element

[0100] 128 open carrier

[0101] 130 membrane

[0102] 132 Surface

[0103] 134 pores

[0104] 136 Culture medium

[0105] 138 enlarged section

[0106] 140 Electrode arrangement

[0107] 142 Feed

[0108] 144, 144' Contact

[0109] 146, 146' contact point

[0110] 148 connection

[0111] 150 measuring and control equipment

[0112] 152 smartphones

[0113] 154 screen

[0114] 156 Diagram

[0115] 160 Cover

[0116] 162 Derivation

[0117] 164 laminar flow

[0118] 166 electrical conductor arrangement

[0119] 168 submersible electrode

[0120] 210 Method for determining an electrochemical quantity of a cell culture

[0121] 212 Provision step Application step Further provision step Suspension step Recording step

Claims

Patent claims 1. Cell culture membrane insert (120), comprising - an open carrier (128) which is designed to expose a cell culture (112) to at least one gas or aerosol (114), wherein a membrane (130) is introduced into the carrier (128), which membrane has at least one surface (132) which is designed to receive the cell culture (112), wherein the membrane (130) o has pores (134) for the diffusion of nutrient medium (136) through the membrane (130), and o has an electrode arrangement (140) on the at least one surface (132) designed to receive the cell culture (112), wherein the electrode arrangement (140) comprises at least two electrodes, and - at least two contact points (146, 146'), each contact point (146, 146') being configured to establish an electrical connection between one of the electrodes and a respective electrical contact (144, 144') not encompassed by the cell culture membrane insert (120).

2. Cell culture membrane insert (120) according to the preceding claim, wherein the membrane (130) is introduced into an inner side of the carrier (128), wherein the inner side of the carrier (128) further comprises at least two electrical conductor arrangements (166, 166') which are designed to establish, on the outer edge of the carrier (128), the electrical connection between one of the electrodes via the associated electrical conductor arrangement (166, 166') and the associated contact point (146, 146) to the respectively associated electrical contact (144, 144') not encompassed by the cell culture membrane insert (120).

3. Cell culture membrane insert (120) according to one of the preceding claims, wherein the electrode arrangement (140) is configured for spatially resolved detection of at least one electrochemical variable of the cell culture (112) upon application of an electrical voltage or an electrical current to the at least two electrodes.

4. Cell culture membrane insert (120) according to one of the preceding claims, wherein the membrane (130) has a diameter of 2 mm to 100 mm.

5. A device (110) for detecting at least one electrochemical parameter of a cell culture (112) exposed to at least one gas or aerosol (114), comprising: - an open housing (122) which is designed to receive o at least one cell culture membrane insert (120) according to one of the preceding claims, o at least one nutrient medium (136) for the cell culture (112), and o at least one supply (142) for the at least one gas or aerosol (114); - at least two electrical contacts (144, 144'), each of which is configured to establish an electrical connection to the at least two contact points (146, 146') comprised by the cell culture membrane insert (120); and - at least one connection (148) to a measuring and control device (150).

6. Device (110) according to the preceding claim, further comprising at least one discharge line (162) for the at least one gas or aerosol (114).

7. Device (110) according to the preceding claim, further comprising at least one cover (160), wherein the at least two electrical contacts (144, 144') are attached to the at least one cover (160), which are configured to establish the electrical connection with the at least two contact points (146, 146') of the cell culture membrane insert (120).

8. Device (110) according to the preceding claim, wherein the at least one cover (160) further comprises the at least one supply (142) and the at least one discharge line (162) for the at least one gas or aerosol.

9. Device (110) according to one of the preceding claims relating to the device (110), wherein the at least one cell culture membrane insert (120) is replaceable.

10. Device (110) according to one of the preceding claims relating to the device (110), wherein the housing (122) - for simultaneously accommodating a plurality of cell culture membrane inserts (120); - for supplying the cell cultures (112) in the majority of the cell culture membrane inserts (120) with the same or a different nutrient medium (136); and - is arranged to expose the cell cultures (112) in the majority of the cell culture membrane inserts (120) to the same or a different gas or aerosol (H4).

11. Device (110) according to one of the preceding claims relating to the device (110), wherein the at least one electrochemical quantity is selected from an impedance of the cell culture (112) or a transepithelial electrical resistance through the cell culture (112).

12. Device (110) according to one of the preceding claims relating to the device (110), further comprising at least one immersion electrode (168), wherein the at least one immersion electrode (168) is for - introduction into the at least one nutrient medium (136) for the cell culture (112); and - detection of at least one further electrochemical parameter of the cell culture (H2) is set up.

13. A method (210) for detecting at least one electrochemical variable of a cell culture (112) exposed to at least one gas or aerosol (114), comprising the steps: a) (212) providing at least one cell culture membrane insert (120) according to one of the preceding claims relating to the cell culture membrane insert (120); b) (214) applying the cell culture (112) to at least one surface (132) of a membrane (130) of the cell culture membrane insert (120); c) (216) providing at least one nutrient medium (136) for the cell culture (112); d) (218) exposing the cell culture (112) to the at least one gas or aerosol (H4); e) (220) detecting at least one electrochemical variable of the cell culture (112) by means of at least one device (110) according to one of the preceding claims relating to the device (110).

14. The method (210) according to the preceding claim, wherein at least one surface of the cell culture (112) facing away from the membrane (130) is exposed at least temporarily to the at least one gas or aerosol (114) during the detection of the at least one electrochemical variable of the cell culture (112) and remains free of the nutrient medium (136) during this time.

15. Use of a cell culture membrane insert (120) according to one of the preceding claims relating to the cell culture membrane insert (120) for detecting at least one electrochemical variable of a cell culture (112) exposed to at least one gas or aerosol (114).

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