Apparatus for ensuring a sterile environment for the incubation of cell cultures - Patent Application 20070122997

Abstract

The present invention relates to an apparatus for ensuring a sterile environment for the incubation of cell cultures, comprising a heating chamber (100) with a heating element, temperature sensors, humidity sensors, and CO2 sensors, as well as a gas supply with gas connections inside. The apparatus also comprises a culture vessel (110) mounted in the heating chamber (100). The culture vessel has sealable channels (130) for each sensor, which are also sealable gas inlets and outlets (130) connected to gas connections inside the heating chamber (100). The sensors in the heating chamber (100) are positioned to fit into the associated channels (130) of the culture vessel (110). The apparatus also comprises a water reservoir (120) mounted in the culture vessel (110). These three elements, i.e., the heating chamber (100), the culture vessel (110), and the water reservoir (120), can be sterilized separately from each other using the method that is optimal in each case. Since the sensor is not part of the culture vessel (110), the latter can also be decontaminated by high temperature. The fact that the heating chamber (100) is sterilized ensures that the cell culture in the culture vessel (110) cannot be contaminated by the outside and cannot become a source of contamination in the laboratory.

Description

[Technical Field]

[0001] The present invention relates to a device for ensuring a sterile environment for the incubation of cell cultures. Such a device is primarily used for the cultivation of various cell cultures, mainly for medical and research purposes, in which the sterility of all elements that come into contact with the cell cultures and the avoidance of any contamination of these cell cultures are of paramount importance.

[0002] The invention of genetic scissors (known as CRISP / CAS) has opened up the possibility of targeted modification of genetic material. This has led various research groups to quickly develop new cell therapy treatments based on the manipulation of genetic material. In the field of human medical therapy, this will involve personalized medicine, in which patient material (i.e., the patient's living cells) are incubated in cell cultures before and after genetic manipulation. To strictly avoid contamination of this patient material, various devices are required to safely ensure a sterile environment for these cell cultures during the incubation phase. [Background technology]

[0003] Various cell cultures are cultivated in incubators in which a selectable constant temperature (usually 37° C.), an adjustable CO concentration (e.g., 5%), and high relative humidity must prevail. Ideal growth conditions are thus provided in the incubator for any type of cell, i.e., both desirable and undesirable cultures.

[0004] A particularly important component of an incubator is the water supply. Essentially, the water in an incubator has the role of ensuring a relatively high humidity level, close to 100%, so that the aqueous nutrient medium in which the cells of the cell culture are placed does not evaporate, or evaporates very little. To prevent the growth of cell biological contaminants in the water reservoir, substances that are toxic to protozoa are typically added to the water.

[0005] Two problems arise from the above: first, contamination can be carried in with the water and then multiply in the incubator, even if toxic substances are added to the water itself. Second, the combination of relatively high humidity and optimal temperatures for cell growth stimulates any contamination that may be present to an explosive growth that is amplified many times if moisture condenses in the incubator.

[0006] In fact, ideal growth conditions in an incubator not only result in the proliferation of patient cells, but also in the unintentional introduction of foreign cells, so-called impurities or contamination, which can lead to contamination of the actual cell culture, i.e., the patient material, which can then become unusable for treatment and, in the worst case, result in the patient's death as a result of the contamination.

[0007] In order to reduce contamination of the incubator with foreign cells, the interior of a conventional incubator must be sterilized (decontaminated) at regular intervals. For this purpose, the interior of the incubator is exposed to temperatures approaching 200°C for periods of up to several hours. In this way, the cell population present therein is largely killed, but not completely eliminated. It is common for contamination to be reduced, for example, by a factor of 1000.

[0008] Another approach to reducing the risk of contamination is to place samples in individual sealed containers. These containers are usually fitted with gas-permeable membranes, resulting in gas and moisture exchange with the atmosphere in the incubator. The membranes themselves are usually designed as sterile filters that primarily minimize the intrusion of biological contaminants into the interior of the container. However, such containers do not prevent contamination within the incubator itself, and especially do not prevent contamination on the outer surface of such containers. Specifically, contamination may preferentially accumulate on the outer surface of the sterile filter of such containers, and as a result, the introduction of this contamination into the interior of the container cannot be excluded.

[0009] In order to monitor or regulate parameters such as temperature or CO2 concentration in an incubator, various sensors must be installed inside the incubator. However, many of these sensors are sensitive to high temperatures; for example, a typical CO2 sensor cannot withstand temperatures above 160°C. This makes decontamination of an incubator cumbersome, as at least some of the sensors must be removed from the incubator beforehand. In addition, although these sensors cannot be decontaminated themselves, they still must be reinstalled inside the incubator, which at least partially negates the effect of decontaminating the incubator.

[0010] For many current applications in cell biology and medicine, and especially for possible future applications, such as in the treatment of genetic diseases (e.g., tumors, leukemias or inherited diseases), a completely sterile environment in an incubator is essential.

[0011] WO 2015 / 172882 discloses a system intended to increase safety in laboratories and specifically to prevent sample mix-ups and possible contamination by means of mutually self-contained mini-incubators, each designed for only one sample carrier. However, this system does not guarantee complete sterility.

[0012] WO 2011 / 130865 describes an essentially conventionally designed incubator, intended to carry out as many processing steps as possible within the incubator mechanically, i.e., without human intervention, in order to prevent sample contamination. Special devices, such as robotic arms, are provided for this purpose. However, complete sterility cannot be guaranteed even in this case. [Prior art documents] [Patent documents]

[0013] [Patent Document 1] International Publication No. 2015 / 172882 [Patent Document 2] International Publication No. 2011 / 130865 Summary of the Invention [Problem to be solved by the invention]

[0014] It is an object of the present invention to provide a device that allows for improved sterility in the incubation of cell cultures. [Means for solving the problem]

[0015] This object is achieved by the subject matter of the independent claims. Advantageous further embodiments of the object of the independent claims are set out in the dependent claims. The language of all claims is hereby incorporated by reference into this description.

[0016] The use of the singular is not intended to exclude the plural, unless otherwise specified, and vice versa.

[0017] To this end, a device for ensuring a sterile environment for incubation of cell cultures is proposed. The device of the present invention has a sealable and / or sealed heating chamber with at least one heating element. The heating chamber has a temperature sensor, a humidity sensor, and / or a CO2 sensor, and at least one connection on the outside for a gas supply. The connection is connected to a conduit that brings the gas connection to the interior of the heating chamber. The gas supply is typically CO2, but other gases may be provided for specific cell cultures. The device of the present invention also has a sealable and / or sealed culture vessel installed in the heating chamber for holding the cell culture. The culture vessel does not have its own heating system. The culture vessel has a sealable flow path for each of the sensors, which is connected to the atmosphere in the culture vessel. The culture vessel also has at least one sealable gas inlet and / or gas outlet that can be connected to at least one gas connection on the interior of the heating chamber. The sensors of the heating chambers are positioned so that they are introduced into the respective sealable channels of the heating chambers when the culture vessels are inserted into the heating chambers. In this way, the sensors of the heating chambers can provide measurement data regarding conditions in the culture vessels. The device of the present invention further includes a water reservoir disposed in the culture tank.

[0018] With this device, the three elements, the heating chamber, the culture vessel, and the water reservoir, can be sterilized separately from each other using the optimal process for each, thereby achieving a significant improvement in the sterility situation while the cell culture is being incubated. Because the sensor is not installed in the culture vessel, the culture vessel can be decontaminated, for example, by high temperature (e.g., above 200°C) if required. The sterilized heating chamber ensures that the cell culture in the culture vessel cannot be contaminated from the outside and that contamination cannot be caused in the laboratory.

[0019] The sealable gas inlet and / or outlet can be connected to gas ports located inside the warming chamber, allowing for the maintenance of optimal conditions in the culture vessel, which can be controlled by sensors housed within the sealable diffuser(s).

[0020] Possible contamination of the cell culture in the culture vessel by the sensors can be prevented by covering each of said sensors in the warming chamber with a sterile filter.

[0021] Similarly, contamination by the gas supply can be avoided by the fact that at least one connection of the heating chamber for the gas supply, the line connected thereto, or the gas connection connected via this line and provided inside the heating chamber, has a sterile filter.

[0022] For inserting the culture vessel into the heating chamber, it is advantageous if the heating chamber has a sealing lid or door, the opening of which has a cross section greater than the width of the culture vessel.

[0023] Contamination by the sensors can also be prevented by ensuring that each sealable flow path leading to the culture vessel has a sterile filter. This design is particularly preferred. This design can be used as a replacement for or a complement to a sterile filter directly in contact with the sensor of the warming chamber.

[0024] Contamination by the gas supply can preferably be prevented by at least one sealable gas inlet and / or gas outlet in the fermentor with a sterile filter, which can consequently be used either directly as a replacement for a sterile filter in the gas connection of the heating chamber or as a complement thereto.

[0025] To ensure that the culture vessel is sterilized before use, it is advantageous if the culture vessel is a disposable element, which can already be manufactured under aseptic or clean room conditions.

[0026] The introduction of the sample carrier(s) containing the cell culture is facilitated if the culture vessel has a sealing lid or door.

[0027] It is particularly advantageous if the water reservoir is covered with a semi-permeable membrane. This membrane must be impermeable to water but allow water vapor to pass through. For example, nonwoven fabrics made from high-density polyethylene (PE-HD) have proven to be excellent for this purpose. In this way, water can be added as soon as the reservoir is made.

[0028] Premature evaporation of water from the reservoir can be prevented if the semi-permeable membrane of the reservoir is sealed before use with a film that is impermeable to water and water vapor, for example, a plastic film, which is removed for use.

[0029] Possible contamination of the cell culture via the water reservoir is particularly unlikely if the reservoir contains sterile, ultrapure water.

[0030] The sterility of the reservoir can be particularly well ensured if the reservoir is a disposable element.

[0031] The incubation conditions in the culture tank can be set particularly optimally if there is a control and / or regulation system that can influence the conditions in the culture tank based on data from sensors in the heating chamber. Data from the temperature sensor is used to control and / or regulate the heating element of the heating chamber, data from the CO2 sensor is used to control and / or regulate at least one gas supply. Data from the humidity sensor is used to output a warning message that can prompt the user to update the water reservoir, for example, if the humidity reaches a value that is too low.

[0032] The problem is further solved by a sealed and / or sealable culture vessel for holding a cell culture. The culture vessel has at least one sealable flow path for a sensor communicating with the atmosphere in the culture vessel. Furthermore, the culture vessel has at least one sealable gas inlet and / or gas outlet connectable to a gas port. Finally, a location for a water reservoir is defined inside the culture vessel.

[0033] Such a culture vessel can be easily decontaminated and / or even manufactured under sterile conditions. Therefore, contamination-sensitive samples can be placed in such a culture vessel, and the contamination-sensitive samples are then specially protected throughout the culture process. The culture vessel can then be placed in a heating chamber as described above. A sealable gas inlet and / or gas outlet can be connected to a gas connection provided inside the heating chamber, allowing optimal conditions to be maintained in the culture vessel. The optimal conditions can be controlled by a sensor located in the sealable channel(s).

[0034] This culture vessel can therefore be used in a device as described above.

[0035] Further details and features can be gleaned from the following description of preferred example embodiments together with the drawings. Each feature can be implemented individually or in combination with each other. The possibilities for solving the problem are not limited to the example embodiments. For example, range specifications always include all (unmentioned) intermediate values ​​and all conceivable subintervals. [Brief explanation of the drawings]

[0036] An example of one embodiment is illustrated diagrammatically in the drawings, in which the same reference numbers in the different figures indicate identical or functionally identical elements or elements that correspond to each other in terms of their functionality.

[0037] [Figure 1] 1 is a schematic overview of a device according to the invention; [Figure 2] Schematic diagram of a warming chamber of a device according to the invention. [Figure 3] Schematic diagram of a culture vessel of a device according to the invention. [Figure 4] Schematic diagram of a reservoir of a device according to the invention. DETAILED DESCRIPTION OF THE INVENTION

[0038] 1 shows a device for ensuring a sterile environment for the incubation of cell cultures in an overall view so that the arrangement of the three main components in relation to one another can be seen. The heating chamber 100 is the outer element and, in normal practice, houses a culture vessel 110, which can be of various sizes depending on the culture to be cultivated. A design in which the culture vessel is able to accept a sample carrier (e.g., a microplate or a Petri dish) is preferred. However, larger designs are of course also possible.

[0039] Disposed within the culture vessel is a water reservoir 120. In selected embodiments, connections or elements 130 are disposed behind the culture vessel, connecting the culture vessel 110 to the warming chamber 100. These connections or elements are described in more detail below in conjunction with Figures 2 and 3.

[0040] 2 shows a heating chamber 100, which forms an outer shell that ensures the desired temperature. For this purpose, the heating chamber houses a heating element 210. Required sensors, for example, sensors for temperature, relative humidity, or CO2 content, are also installed in the heating chamber 100.

[0041] In Figure 2, for clarity, only CO2 sensor 220 is shown. The sensor is mounted in a holder 230, which positions the sensor so that it is in the correct place when the culture vessel is inserted. Furthermore, at least one connection for a gas supply is provided. This port is connected to a conduit that leads to a gas port 240 inside the warming chamber. Preferably, two such gas ports are provided, one of which provides, for example, a CO2 supply and the other a means for gas extraction.

[0042] Preferably, a door 250 designed to be airtight is provided opposite the sensor and gas connection. Alternatively, for example, a removable lid or the like may be provided. The opening of the lid or door 250 must have a cross section large enough to allow the culture vessel 110 to be inserted without difficulty.

[0043] To avoid contamination of the laboratory environment by bringing the warming chamber 100 into the laboratory, it is advantageous if the warming chamber is cleaned and placed in a packaging container that can sterilize the warming chamber before being brought into the laboratory. The packaging consists at least in part of a semi-permeable membrane that is gas permeable but has pores that are sufficiently fine that bacteria cannot pass through. Sterilization can then be carried out in a known manner, for example, by gassing with ethylene oxide.

[0044] FIG. 3 shows a culture vessel 110 that can be inserted into and removed from a warming chamber. The culture vessel has at least one sealable flow path 310 for a sensor, which is in communication with the atmosphere in the culture vessel. This flow path 310 is preferably lined on the inside with a sterile filter 320, thereby protecting the culture vessel from external contamination. Each sensor in the warming chamber is positioned to be inserted into a respective sealable flow path of the culture vessel 110 when the culture vessel 110 is inserted into the warming chamber. As a result, the center can provide data regarding the condition inside the culture vessel 110. The culture vessel also includes at least one sealable gas inlet and / or gas outlet 330 that is adapted to at least one gas port inside the warming chamber. Each of these gas inlets and / or outlets preferably has a sterile filter 340, thereby protecting the culture vessel 110 from external contamination but also preventing contamination from the culture vessel to the environment. This may be necessary, for example, if infectious agents are present in the cell culture. Preferably, a gas inlet and a gas outlet are provided in each case, which makes it easier to carry out gas exchange inside the culture vessel.

[0045] Furthermore, the culture vessel 110 has an opening through which, for example, a sample carrier containing a cell culture can be inserted into or removed from the culture vessel. This opening can be closed by a lid or door 350. The closure by the lid or door 350 should ensure that the culture vessel 110 is hermetically sealed against the environment. The interior of the culture vessel is further provided to receive a reservoir 120.

[0046] To achieve the goal of ensuring a sterile environment for the incubation of cell cultures, it is provided that the culture vessel 110 is cleaned and sterilized prior to its use. In this regard, it is advantageous if the culture vessel 110 is disposable. The elements that are particularly critical for contamination, namely the sensor flow path 310, the gas inlet or outlet 330, and the associated sterile filters 320 / 340, are part of the culture vessel. Therefore, they are included in the delivery, sterilized, and discarded.

[0047] It also makes sense to provide special packaging, for example, a semi-permeable membrane, for sterilizing the culture vessel 110. Various methods can be used for sterilization, for example, gamma irradiation, or gas treatment with ethylene oxide, or thermal methods can be used.

[0048] In order to use the culture vessel 110 in a clean room environment, it is likewise advantageous if the entire manufacture of the culture vessel, including the sterile packaging, is itself carried out in a clean room. The finished component (i.e., the culture vessel 110 in its sterile packaging) can then still be provided with a clean room compatible outer packaging (e.g., a plastic bag).

[0049] In Figure 4, a water reservoir 120 is shown diagrammatically as a further component of the device according to the invention. In fact, it does not necessarily have to be rectangular. For example, cylindrical or bubble-shaped designs are also conceivable. The water reservoir 120 preferably has a semi-permeable membrane 410, for example a Tyvek film or similar, on at least part of its surface. The semi-permeable membrane allows gaseous water vapor to escape from the reservoir to the atmosphere in the culture vessel, but prevents liquid water from leaking out of the reservoir.

[0050] Before the reservoir is inserted into the culture vessel, the semi-permeable membrane is preferably sealed with a film 420, for example, a watertight and airtight plastic film. The arrow in Figure 4 indicates that this sealing film 420 is removed before the reservoir is inserted.

[0051] It is particularly advantageous if the water reservoir 120 is designed as a separate disposable reservoir to be inserted into the culture vessel 110. In this case, the water reservoir 120 can be filled with water as soon as possible during manufacture. Preferably, sterile, ultrapure water is used.

[0052] To achieve the goal of providing a sterile environment for incubation, it is advantageous if the reservoir 120 is externally sterilized prior to its use. To this end, the reservoir can be placed in a sterilization packaging, for example, a packaging container with a semi-permeable membrane, in which the other components, i.e., the heating chamber 100 and the culture vessel 110, can be placed. Various methods can be used for sterilization, for example, gamma irradiation or gassing with ethylene oxide.

[0053] For use of the water reservoir 120 in a clean room environment, it is also advantageous if the entire manufacturing of the water reservoir, including the water and sterile packaging, is carried out in a clean room and the entire components (i.e., the water reservoir 120, the water, the water vapor permeable membrane 410, the seal 420, and the sterile packaging) are repackaged in a clean room suitable container (e.g., a plastic bag).

[0054] The above described device provides the following advantages over the prior art:

[0055] Contamination of the sterile filter from the area of ​​the warming chamber is not possible because the gas inlet of the culture vessel is fitted to the warming chamber in such a way that there is no gas exchange between the warming chamber and the culture vessel. The gas inlet or gas outlet only allows gas to pass from the gas connection to the culture tank.

[0056] Similarly, the gas outlet of the culture vessel is fitted to the warming chamber in such a way that gas exchange between the culture vessel and the warming chamber is prevented, in this way contamination of the warming chamber from the region of the culture vessel cannot occur.

[0057] Furthermore, the device according to the invention has the advantage that the sensor is placed outside the culture vessel and therefore cannot be contaminated by the sample in the culture vessel since it is separated from the culture vessel by a sterile filter.

[0058] Additionally, the use of sterile culture vessels prevents contamination of the sample, especially if the culture vessels are sterile, disposable items.

[0059] The use of ultrapure water also eliminates contamination of the introduced water because the ultrapure water is introduced to the sterile culture vessel in a sterile, disposable reservoir. This risk does not exist when the relative humidity is so high that water condenses in the culture vessel because all components are sterilized before use.

[0060] term incubator An incubator is a device used in biology to create and maintain controlled outdoor conditions for various developmental and growth processes. Incubators are used to create and maintain a microclimate in which humidity and temperature conditions are tightly controlled. Incubators are equipped with timers and temperature controllers, and also with settings to regulate the supply of fresh air under certain circumstances. The set temperature is adjusted to the optimum temperature for the microorganisms to be incubated. CO2 incubators are used for the cultivation of animal cells. (according to de.wikipedia.org / wiki / Incubator_(Biology))

[0061] Microplates A microwell plate (or microplate) is a multi-sample carrier. Usually rectangular, microplates are typically made from plastic, although glass is also available for highly specialized applications. They contain between 6 (2 x 3) and 1536 (32 x 48) isolated wells arranged in rows and columns. Their exact dimensions (length x width x height) are 127.76 x 85.48 x 14.35 mm, according to the ANSI standard based on recommendations from the Society for Biomolecular Screening (SBS). Microplates are used for a wide variety of microbiological procedures. Typical applications are cell culture or industrial bioreaction screening. Due to the large number of wells and the uniformity of the wells, microplates are suitable for the parallel incubation and testing of a large number of different samples. Due to their standardized size, nearly all procedures can be automated using suitable robots. (From de.wikipedia.org / wiki / Microtiterplate)

[0062] Sample Carrier Microbiological samples or cell cultures are stored in containers referred to herein as sample carriers. Depending on the type of sample, the sample carrier can be a wide variety of containers. Typically, however, Petri dishes or microplates are used, and sometimes Erlenmeyer flasks or the like are also used. Multiple sample carriers (e.g., microplates) are also referred to as sample carrier systems.

[0063] Sterile Filter In sterile filtration, microorganisms are separated from the material to be sterilized by filtration. Membranes with pore sizes between 0.1 μm and 0.22 μm are usually used as filters. Sterile filtration is often used to sterilize heat-sensitive solutions, for example, tissue culture fluids containing serum. The main applications are the sterile filtration of aqueous solutions, heat-sensitive nutrient solutions, vitamin solutions, serum, viral vaccines, plasma fractions, and protein solutions. (From https: / / de.wikipedia.org / wiki / Sterilisation#Sterilfiltration) [Explanation of symbols]

[0064] 100 Heating Chamber 110 Culture vessel 120 Water Tank 130 Connection or connecting element 210 Heating element 220 CO2 sensor Holder for 230 sensors 240 Gas Connection 250 Heating chamber door 310 Flow path for sensor Sterile filter for 320 sensors 330 Gas inlet or gas outlet 340 Sterile filters for gas inlet or gas outlet 350 Culture tank door 410 Semi-permeable membrane 420 film

[0065] References International Publication No. 2015 / 172882 International Publication No. 2011 / 130865

Claims

1. 1. A device for providing a sterile environment for incubation of cell cultures, comprising: Sealed heating chamber (100) however, The heating chamber (100) includes at least one heating element (210); The heating chamber (100) includes a temperature sensor and / or a humidity sensor and / or a CO2 sensor (220); The heating chamber (100) has at least one connection on the outside for a gas supply, said connection being connected to a line that brings a gas connection (130) into the interior of the heating chamber; a sealed culture vessel (110) for holding said cell culture; however, The culture vessel (110) is placed in the heating chamber (100); The incubation vessel (110) has a sealable flow path (310) for each of the sensors in the heating chamber, the flow path communicating with the atmosphere in the incubation vessel; The culture vessel (110) includes at least one sealable gas inlet and / or gas outlet (330) connectable to at least one gas port (240) inside the warming chamber; The sensors of the heating chamber (100) are positioned so as to be introduced into the sealable channels (310) of each of the culture vessels (110) when the culture vessels (110) are inserted into the heating chamber (100); and Water tank (120) however, The reservoir (120) is disposed in the culture vessel (110); Devices containing:

2. Each of the sensors in the heating chamber (100) is covered by a sterile filter.

2. The device according to claim 1, characterized in that:

3. The at least one connection (240) of the heating chamber for gas supply, the conduit connected thereto, or the gas connection (130) connected via this conduit and provided inside the heating chamber, contains a sterile filter.

3. A device according to claim 1 or 2, characterized in that it comprises:

4. the heating chamber (100) having a sealing lid or door (250); The opening of the lid or door (250) has a cross section greater than the width of the culture vessel (110). A device according to any one of claims 1 to 3, characterized in that it

5. Each sealable channel (310) leading to the culture vessel (110) includes a sterile filter (320). A device according to any one of claims 1 to 4, characterized in that it

6. The at least one sealable gas inlet and / or gas outlet (330) in the culture vessel (110) includes a sterile filter (340). A device according to any one of claims 1 to 5, characterized in that it

7. The culture vessel (110) is a disposable element. A device according to any one of claims 1 to 6, characterized in that it

8. The culture vessel (110) has a sealing lid or a sealing door (350). A device according to any one of claims 1 to 7, characterized in that it

9. The reservoir (120) is covered by a semi-permeable membrane (410). A device according to any one of claims 1 to 8, characterized in that it

10. The semi-permeable membrane (410) of the water reservoir (120) is sealed before use with a film (420) that is impermeable to water and water vapor, which is removed for use.

10. The device according to claim 9, characterized in that

11. The water reservoir (120) contains sterile, ultra-pure water. Device according to any one of claims 1 to 10, characterized in that it

12. The water reservoir (120) is a disposable element A device according to any one of claims 1 to 11, characterized in that it

13. A control and / or regulation system is provided; however, The control and / or regulation can affect conditions in the culture vessel (110) based on data from the sensor (220) of the heating chamber (100); The temperature sensor data is used to control and / or regulate the heating element (210) of the warming chamber (100); and / or the data of the CO2 sensor (220) serves to control and / or regulate the at least one gas supply; and / or The data of the humidity sensor is useful for outputting a warning message. Device according to any one of claims 1 to 12, characterized in that it

14. A sealed culture vessel (110) for holding a cell culture, comprising: At least one sealable channel (310) for the sensor however, the at least one sealable channel (310) is in communication with the atmosphere in the culture vessel (110); at least one sealable gas inlet and / or gas outlet (330) connectable to the gas port (240); and A location for a water reservoir (120) inside the culture vessel (110) A culture vessel (110) comprising:

15. A culture vessel (110) according to claim 14 for use in a device according to any one of claims 1 to 13.

16. Use of a culture vessel (110) according to claim 14 in a device according to any one of claims 1 to 13.

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