Drying prevention equipment and cell culture kit

Abstract

To provide drying inhibiting instruments that can inhibit the evaporation of water from the medium held by the well plate, and to provide cell culture kits that have such a drying inhibiting instrument and enable suitable cell culture.SOLUTION: Provided is a drying inhibiting instrument comprising: a main body that houses a well plate, and a lid provided on the main body in an openable manner, the main body having a holding part that holds the well plate, a liquid storage tank that stores liquid, and a support part that supports the holding part apart from the bottom surface inside the liquid storage tank, an internal space surrounded by the liquid storage tank and the lid being in communication with the outside in the condition that the lid is closed.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] The present invention relates to a drying prevention instrument and a cell culture kit. [Background technology]

[0002] In recent years, technological developments have been made to replace some of the clinical tests that were previously conducted on animals with in vitro tests using live cells. In particular, with the advancement of stem cell technology, studies are being made on tests using human-derived stem cells as live cells.

[0003] In an in vitro test using live cells, for example, live cells and culture solution are placed in the wells of a well plate, and the drug to be evaluated is then added and the live cells are cultured. The shape, intercellular connections, etc. of the cultured live cells are evaluated to evaluate the effects of the drug on the live cells.

[0004] Generally, well plates are also called microplates or microtiter plates depending on the size of the wells and the purpose of use. In this specification, the term "well plate" will be used consistently.

[0005] It is known that in such in vitro tests, when cells are cultured for a long period of time, water evaporates from the culture medium, which causes fluctuations in the drug concentration in the culture medium and affects cell culture.

[0006] It is also known that water evaporation from the culture solution is likely to occur in wells close to the periphery of the well plate among the multiple wells arranged in a matrix on the well plate. Therefore, when water evaporates from the culture solution, the concentration of the culture solution differs between the wells located near the periphery of the well plate and the wells located near the center of the well plate, which may cause variations in cell culture.

[0007] In response to such problems, a well plate has been proposed that has compartments outside the rows of wells for holding a liquid for regulating evaporation (see, for example, Patent Document 1). Summary of the Invention [Problem to be solved by the invention]

[0008] However, well plates generally have a size that conforms to the SBS standard. In the well plate described in Patent Document 1, it is necessary to secure an area for forming a required number of wells in a well plate of a standardized size, and form a partition for holding liquid in the remaining area. Therefore, in the well plate described in Patent Document 1, the amount of liquid for regulating evaporation that can be stored is limited to a small amount, making it difficult to suppress water evaporation from the medium when cell culture is continued for a long period of time.

[0009] In recent years, well plates have become more and more densely packed, and compared to pre-packaged well plates, the amount of culture medium stored in each well has decreased. As a result, the adverse effects of water evaporation from the culture medium have become more pronounced, and measures to address this issue have been required.

[0010] The present invention has been made in view of the above circumstances, and aims to provide a desiccation prevention device capable of preventing water evaporation from a culture medium held in a well plate, and a cell culture kit including such a desiccation prevention device and enabling suitable cell culture. [Means for solving the problem]

[0011] In order to solve the above problems, one aspect of the present invention provides a dryness suppression device comprising a main body for accommodating a well plate and a lid provided on the main body that can be opened and closed freely, the main body having a holding portion for holding the well plate, a liquid storage tank for storing liquid, and a support portion for supporting and spacing the holding portion from the inner bottom surface of the liquid storage tank, and when the lid is closed, the internal space surrounded by the liquid storage tank and the lid is connected to the outside. Effect of the Invention

[0012] According to the present invention, it is possible to provide a desiccation prevention device capable of preventing water evaporation from a culture medium held in a well plate, and a cell culture kit including such a desiccation prevention device and enabling suitable cell culture. [Brief description of the drawings]

[0013] [Figure 1] FIG. 1 is a schematic perspective view showing a drying prevention instrument 1 and a cell culture kit 100 according to an embodiment. [Diagram 2] FIG. 2 is a cross-sectional view taken along line II-II in FIG. [Diagram 3] FIG. 3 is an explanatory diagram of a drying prevention device 1X not having the configuration of the present invention and a cell culture kit 100X having the drying prevention device 1X. [Figure 4] FIG. 4 is an explanatory diagram of the dryness prevention device 1 according to the embodiment and a cell culture kit 100 having the dryness prevention device 1. As shown in FIG. [Diagram 5] FIG. 5 is an explanatory diagram of a drying prevention tool 2A and a cell culture kit 200A according to a second embodiment of the present invention. [Figure 6] FIG. 6 is an explanatory diagram showing a dryness prevention tool 2B and a cell culture kit 200B according to a modified example of the second embodiment. [Figure 7] FIG. 7 is an explanatory diagram of a drying prevention tool 3A and a cell culture kit 300A according to a third embodiment of the present invention. [Figure 8] FIG. 8 is an explanatory diagram showing a dryness prevention tool 3B and a cell culture kit 300B according to a modified example of the third embodiment. [Figure 9] FIG. 9 is an explanatory diagram of a dryness prevention tool 4 and a cell culture kit 400 according to a fourth embodiment of the present invention. [Figure 10] FIG. 10 is an explanatory diagram of a dryness prevention tool 5 and a cell culture kit 500 according to a fifth embodiment of the present invention. [Figure 11]FIG. 11 is an explanatory diagram of a drying prevention tool 6 and a cell culture kit 600 according to a sixth embodiment of the present invention. [Figure 12] FIG. 12 is an explanatory diagram showing the results of the example. [Figure 13] FIG. 13 is an explanatory diagram showing the results of the example. [Figure 14] FIG. 14 is an explanatory diagram showing the results of the example. [Figure 15] FIG. 15 is an explanatory diagram showing the results of the example. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] [First embodiment] Hereinafter, a drying prevention device and a cell culture kit according to a first embodiment of the present invention will be described with reference to Figures 1 to 4. In addition, in all the following drawings, the dimensions and ratios of each component are appropriately changed to make the drawings easier to see.

[0015] In the following description, an xyz Cartesian coordinate system is set, and the positional relationship of each member may be described with reference to this xyz Cartesian coordinate system. In this specification, a specific direction in a horizontal plane is defined as the x-axis direction, a direction perpendicular to the x-axis direction in the horizontal plane is defined as the y-axis direction, and a direction perpendicular to both the x-axis direction and the y-axis direction (i.e., the vertical direction) is defined as the z-axis direction.

[0016] In addition, when describing the positions of components relative to one another, a component located in the +z-axis direction may be referred to as "above" or "above" another component. Similarly, a component located in the -z-axis direction may be referred to as "below" or "below" another component.

[0017] Fig. 1 is a schematic perspective view showing a drying prevention device 1 and a cell culture kit 100 according to the present embodiment. Fig. 2 is a cross-sectional view taken along line II-II in Fig. 1.

[0018] <Drying suppression equipment> The drying prevention device 1 has a main body 10 and a lid 20. The drying prevention device 1 is used to prevent water evaporation from a culture medium filled in a well plate 90 when cells contained in the well plate 90 are cultured.

[0019] In this specification, the term "medium" includes both liquid medium (culture solution) and gel medium. The term "gel medium" refers to a medium used in cell culture, which is the technical field of this embodiment, and is semi-solidified to the extent that it has lost fluidity. Examples of "gel medium" include agar medium and hydrogel medium.

[0020] (Main body) The main body portion 10 has a liquid storage tank 11 , a holding portion 12 , and a support portion 13 .

[0021] The liquid storage tank 11 is a container having an opening 11a that opens upward, and accommodates a holding part 12 and a support part 13 in the internal space. More specifically, the liquid storage tank 11 has a bottom plate 111 and a side wall 112 that is provided on the periphery of the bottom plate 111 and extends upward. The liquid storage tank 11 shown in Fig. 1 is a thin container that is rectangular in plan view.

[0022] The liquid storage tank 11 stores, in an inner space surrounded by a bottom plate 111 and a side wall 112, a liquid L for suppressing evaporation of water from the culture medium filled in the well plate 90. The "liquid for suppressing evaporation of water from the culture medium" corresponds to the "liquid for suppressing drying" in this invention.

[0023] The liquid L is preferably a liquid that does not inhibit the growth of cells in the well 911, and is preferably pure water.

[0024] Furthermore, in order to suppress the growth of bacteria in the liquid L, a preservative or an antibiotic may be added to the liquid L. For example, the liquid L may be an aqueous solution in which a preservative or an antibiotic is added to pure water in an amount that does not inhibit the growth of cells in the well 911. The content of the preservative in the liquid L or the content of the antibiotic in the liquid L may be determined by conducting a preliminary experiment in advance to confirm that the growth of cells in the well 911 is not inhibited.

[0025] The holder 12 is a member that holds the well plate 90. The holder 12 has an area 12a for holding the well plate 90. The holder 12 shown in Fig. 1 is a thin container that is rectangular in plan view, and has a bottom plate 121 on which the well plate 90 is placed, and a side wall 122 that surrounds the area 12a and extends upward. The area 12a is set to be slightly larger than the shape of the well plate 90 in plan view that complies with the SBS standard, so that the well plate 90 can be placed thereon without being impacted by the side wall 122.

[0026] The holding part 12 has the side wall 122, which facilitates positioning of the well plate to be placed thereon. In addition, when the well plate 90 is placed on the holding part 12, the well plate 90 is less likely to move on the holding part 12, which facilitates handling.

[0027] It should be noted that the holding portion 12 does not need to have the side wall 122 as long as the region 12a is secured. As described above, the side wall 122 functions as a structure that defines the region 12a, but the structure that defines the region 12a is not limited to this. When the holding part 12 does not have the side wall 122, a mark in the form of a depression or a protrusion may be provided on the surface of the bottom plate 121 as a structure that indicates the region 12a.

[0028] The periphery of the region 12a in the holding portion 12, i.e., the side wall 112 of the liquid storage tank 11 and the side wall 122 of the holding portion 12 are spaced apart in a plan view. The gap between the side wall 112 and the side wall 122 functions as a steam port 10a. The liquid L stored in the liquid storage tank 11 is exposed from the steam port 10a. The vapor of the liquid L stored in the liquid storage tank 11 is released from the liquid storage tank 11 through the steam port 10a.

[0029] The support parts 13 support the holding part 12 by lifting it away from the inner bottom surface 11x of the liquid storage tank 11. The support parts 13 shown in Fig. 1 are columnar members provided at the four corners of the holding part 12 in a plan view.

[0030] The support parts 13 are provided integrally with the bottom plate 121 of the holding part 12 at the four corners of the holding part 12. The support parts 13 are also provided integrally with the liquid storage tank 11. That is, the main body part 10 of this embodiment is a single member in which the liquid storage tank 11, the holding part 12, and the support parts 13 are provided integrally.

[0031] (lid) The lid 20 is a member that is provided on the main body 10 so as to be freely opened and closed, and that covers the opening 11a. The lid 20 is configured so as to be freely attached and detached to the main body 10. More specifically, the lid 20 has a ceiling plate 21 and a side wall 22 that is provided on the peripheral portion of the ceiling plate 21 and extends downward. The lid 20 fits into the main body 10 on the inner peripheral side of the side wall 22.

[0032] The lower surface 21a of the ceiling plate 21 contacts the upper end 112a of the side wall 112 of the liquid storage tank 11, and determines the height direction position of the lid 20. The distance W between the lower surface 21a and the upper surface 90a of the well plate 90 is preferably, for example, not less than 0.1 mm and not more than 20 mm.

[0033] The lid 20 covers at least the steam port 10a in a plan view, and also covers the opening 11a from the side wall 112 of the liquid storage tank 11 to the region 12a.

[0034] In the present embodiment, the lid 20 is detachable from the main body 10, but this is not limiting. For example, the lid 20 may be configured to be connected to the main body 10 via a hinge, and the lid 20 may be provided so as to be openable and closable.

[0035] The lid 20 has a ventilation hole 20a in the ceiling plate 21. The ventilation hole 20a connects the internal space surrounded by the main body 10 and the lid 20 to the atmosphere outside the dryness suppression device 1. This allows the dryness suppression device 1 to take in gas from the external atmosphere into the internal space through the ventilation hole 20a. In addition, the dryness suppression device 1 can exhaust gas from the internal space to the outside through the ventilation hole 20a.

[0036] In addition, the "internal space surrounded by the main body 10 and the lid 20" refers to the space surrounded by the main body 10 and a virtual lid 20 that does not have ventilation holes 20a, assuming that the lid 20 shown in Figure 1 does not have ventilation holes 20a.

[0037] Moreover, the "outside" refers to the outside of the space partitioned by the main body 10 and the imaginary lid 20 that does not have the ventilation hole 20a.

[0038] The ventilation hole 20a is preferably located at a position overlapping with the region 12a of the holding portion 12 in a plan view. In Figures 1 and 2, the ventilation hole 20a is shown as a through hole located at a position overlapping with the vicinity of the center of the region 12a of the ceiling panel 21.

[0039] In addition, it is preferable that the ventilation hole 20a is not provided with a structure that is considered to hinder ventilation, such as a filter. The environment inside the incubator is usually suppressed from dust and dirt. Furthermore, since the well plate itself has a lid, the intrusion of external factors that may affect the results of cell culture is suppressed. Therefore, it is not necessary to provide a filter in the ventilation hole 20a.

[0040] In general, the pH of the culture medium during cell culture is preferably within the optimum pH range. The optimum pH is, for example, pH 6.8 to 7.2. A typical culture medium is adjusted to have the optimum pH when the CO2 concentration in the environment is 5 to 10%.

[0041] On the other hand, if the culture medium is exposed to air, the medium becomes alkaline. Also, as the cultured cells metabolize, lactic acid is excreted into the medium, making the medium acidic.

[0042] To cope with such fluctuations in the pH of the culture solution, the carbon dioxide concentration in the ambient atmosphere is usually adjusted to 5-10% in an incubator where cells are cultured, and the pH of the culture solution is adjusted. When culturing cells in wells 911 of well plate 90, it becomes necessary to deliver gas with a carbon dioxide concentration adjusted to the above value to the culture solution in wells 911. In the following explanation, the gas with a carbon dioxide concentration adjusted to the above value is referred to as "adjusted gas G."

[0043] The drying suppression device 1 can take in the conditioning gas G from the surrounding environment through the ventilation holes 20a, and can prepare the culture environment for the cells placed on the well plate 90.

[0044] In the dryness suppression device 1, in an atmosphere of adjusted gas G with a CO2 concentration of 5%, the inflow rate of adjusted gas G from the atmosphere (surrounding environment) to the internal space is preferably 20 ppm / sec or more. The humidity of the atmosphere is, for example, 90% (relative humidity, @37°C).

[0045] The inflow rate of the conditioning gas G is more preferably 50 ppm / sec, and even more preferably 100 ppm / sec.

[0046] The inflow rate of the conditioning gas G is not particularly limited as long as it does not inhibit cell culture in the well plate, but is preferably substantially 10,000 ppm / sec or less.

[0047] The upper and lower limit values ​​of the inflow rate of the adjustment gas G can be combined in any desired manner.

[0048] The inflow rate of the adjustment gas G is measured by placing an NDIR (non-dispersive infrared) CO2 concentration sensor (HJ-CO2-LOG, manufactured by Sato Shoji Co., Ltd.) in the holder of the dryness suppression device, and placing the dryness suppression device in an atmosphere with a CO2 concentration of 5% and a humidity of 90% (relative humidity, @37°C). The "rate of increase in carbon dioxide concentration per unit time" obtained from the carbon dioxide measurement value by the CO2 concentration sensor is the desired "carbon dioxide inflow rate."

[0049] The inflow speed of the adjustment gas G can be adjusted by controlling the position of the ventilation hole 20a and the opening area of ​​the ventilation hole 20a. The opening area of ​​the ventilation hole 20a can be adjusted by controlling the opening diameter of the ventilation hole 20a and the number of the ventilation holes 20a.

[0050] In this case, the culture conditions of the cells placed inside the wells 911 to which the conditioning gas G taken in through the ventilation holes 20a is easily supplied may be different from those of the wells 911 to which the conditioning gas G is not easily supplied. In contrast, as shown in Figures 1 and 2, when the ventilation holes 20a are positioned so as to overlap with the center of the region 12a, the conditioning gas G taken in through the ventilation holes 20a is likely to be evenly distributed among the multiple wells 911 of the well plate 90, making it easier to control the culture conditions.

[0051] In the dryness suppression device 1 of the present embodiment, the ventilation hole 20a is a through hole provided in the lid 20, but is not limited thereto. The ventilation hole 20a can be provided in at least one of the lid 20 or the liquid storage tank 11, and various configurations can be adopted, so long as the configuration has a function of introducing the adjusted gas G in the incubator into the dryness suppression device 1. Also, a tubular configuration for introducing the adjusted gas G into the dryness suppression device 1 may be provided in communication with the ventilation hole 20a.

[0052] Moreover, it is preferable that the dryness prevention device 1 has a residual water rate of 75% or more as determined by the following measurement method. (Measurement method) An SBS standard 96-well plate in which all wells are filled with 200 μL of water is placed in the main body, water is stored in the storage tank, and the plate is left in the state in which the lid is closed. After leaving it for 20 days, the rate of change in the amount of water relative to the initial value is calculated based on the following formula (1). (Water Residual Rate)(%)=B / A×100 …(1) (A shows the average initial water volume in four wells located at the four corners of the well plate. B shows the average amount of water remaining in the four wells located at the four corners of the well plate.)

[0053] More specifically, all wells of an SBS standard 96-well plate are filled with 200 μL of water, and then the well plate is covered with a lid that comes with the well plate. The liquid volume in the wells at the four corners of the well plate is measured, and the arithmetic mean value of the water volume in the wells at the four corners is calculated. The calculated arithmetic mean value is used as the initial value.

[0054] Pure water is stored in the storage tank, and the covered well plate is placed on the holder of the drying prevention device. The drying prevention device is then covered with a lid and kept in an incubator at a temperature of 37°C and a humidity of 90% or more.

[0055] After 20 days, measure the liquid volume in the wells at the four corners of the well plate and calculate the arithmetic mean of the liquid volume in the wells at the four corners. Calculate the rate of change of the liquid volume from the initial value using the above formula (1).

[0056] The amount of liquid in the wells is measured using a plate reader. A calibration curve is prepared in advance based on the relationship between the absorbance of the 970 nm laser light and the amount of liquid in the wells, and the amount of liquid in the wells is calculated from the absorbance of water in each well measured using the plate reader.

[0057] The residual rate is preferably 80% or more, and more preferably 90% or more. The upper limit of the residual rate is ideally 100%.

[0058] Use of such a drying suppression device 1 is expected to prevent drying of the outer periphery of a well plate. This reduces the difference in culture environment between the peripheral wells and the central wells of the well plate used, and suppresses variation in culture results.

[0059] (Well plate) The well plate 90 used is a well plate having a configuration that is typically used in cell culture, which is the technical field of this embodiment. The well plate 90 shown in Figures 1 and 2 has a plate body 91 in which multiple wells 911 are provided, and a lid 92 that covers the plate body 91. The well plate 90 that can be used has a size that complies with the SBS standard.

[0060] The multiple wells 911 contain cells (not shown) to be cultured and a medium CM used for culturing the cells.

[0061] The plate body 91 has a peripheral edge provided with a step 91a that protrudes toward the outer periphery.

[0062] The lid 92 is detachably attached to the plate body 91 and is a member that covers the multiple wells 911 from above. More specifically, the lid 92 has a ceiling plate 921 and a side wall 922 that is provided on the periphery of the ceiling plate 921 and extends downward. The lid 92 fits into the plate body 91 on the inner periphery of the side wall 922. A lower end 922a of the side wall 922 contacts a step 91a of the plate body 91, and determines the position of the lid 92 in the height direction.

[0063] <Cell culture kit> The cell culture kit 100 includes a drying prevention device 1 and a well plate 90 housed in the drying prevention device 1. The well plate 90 includes a plurality of wells 911, and has cells (not shown) housed inside the wells 911 and a culture medium (not shown) filled inside the wells 911.

[0064] The cell culture kit 100 is placed in, for example, a known incubator and used to culture the cells contained in the wells 911 .

[0065] When culturing cells in an incubator, if the culture period becomes long, the culture medium in the well 911 may dry out, and the culture conditions of the cells placed in the well 911 may deviate from the desired conditions. In contrast, in the cell culture kit 100 of this embodiment, the well plate 90 is housed in the above-mentioned drying suppression device 1, and the drying of the culture medium in the well 911 can be suppressed.

[0066] 3 and 4 are diagrams for explaining the effect of the dryness prevention device of this embodiment, and are partially enlarged views showing a cross section of the peripheral edge part of the dryness prevention device.

[0067] Fig. 3 is an explanatory diagram of a dryness prevention instrument 1X not having the configuration of the present invention and a cell culture kit 100X having the dryness prevention instrument 1X. Fig. 4 is an explanatory diagram of a dryness prevention instrument 1 of the present embodiment and a cell culture kit 100 having the dryness prevention instrument 1.

[0068] The dryness prevention device 1 and the dryness prevention device 1X differ in the size of the area covered by the lid. 3, an end A of a ceiling plate 21X on the ventilation hole side overlaps with the steam port 10a in a plan view. That is, the lid 20X does not cover the opening 11a from the side wall 112 of the liquid storage tank 11 to the region 12a of the holder 12 that holds the well plate 90.

[0069] 4, the end B on the ventilation hole side of the ceiling plate 21 of the lid 20 of the dryness suppression device 1 overlaps with the region 12a in a plan view. That is, as described above, the lid 20 covers the steam port 10a in a plan view, and also covers the opening 11a from the side wall 112 of the liquid storage tank 11 to the region 12a in the holder 12 that holds the well plate 90.

[0070] The above-mentioned differences in configuration result in the following differences in effects.

[0071] First, as shown in FIG. 3, the well plate 90 is heated, for example, in an incubator (not shown), which promotes evaporation of water from the medium CM in the wells 911, generating steam V1.

[0072] In the dryness prevention device 1X, evaporation of the liquid L stored in the liquid storage tank 11 is promoted by heating in an incubator (not shown), and the generated vapor V reaches the lid 20X. Here, in the dryness prevention device 1X, the lid 20X does not cover the opening 11a until it reaches the region 12a, so the vapor V further diffuses and spreads from the ventilation holes in the lid 20X.

[0073] Therefore, the peripheral portion α of the region 12a in plan view has low humidity. As a result, moisture evaporated from the medium CM in the wells 911 is released to the peripheral portion α through the gap between the plate body 91 and the lid 92 of the well plate 90, and the medium CM dries out. If the medium CM dries out, the cell culture conditions will differ between the wells 911, which may make it difficult to perform an appropriate in vitro test.

[0074] 4, evaporation of the liquid L stored in the liquid storage tank 11 is promoted by heating in an incubator (not shown). The generated steam V forms an airflow that flows upward from the steam port 10a and reaches the lid 20. In the dryness suppression device 1, the lid 20 covers the opening 11a until it reaches the region 12a, so that the airflow of the steam V spreads along the lower surface 21a of the lid 20 (ceiling panel 21) and fills the peripheral portion α of the region 12a in a plan view.

[0075] Since the size of region 12a is set according to the standard size of well plate 90, peripheral portion α overlaps with the peripheral portion of well plate 90 in plan view. Therefore, in cell culture kit 100 including the dryness suppression device 1, the peripheral portion of well plate 90 is filled with steam V during cell culture. Therefore, peripheral portion α of region 12a in plan view has a higher humidity than that of dryness suppression device 1X.

[0076] In the well plate 90 placed in the drying suppression device 1 having such a configuration, moisture evaporated from the medium CM in the wells 911 is unlikely to diffuse toward the outside of the well plate 90, and drying of the medium CM is suppressed. As a result, differences in cell culture conditions are unlikely to occur between the wells 911, resulting in suitable cell culture and enabling appropriate in vitro testing.

[0077] Furthermore, in the dryness prevention device 1, the holding part 12 is raised by the support part 13, and a space capable of storing the liquid L is also formed below the holding part 12. Therefore, compared to a dryness prevention device in which the holding part 12 is provided directly on the bottom surface of the liquid storage tank 11 and there is no space below the holding part 12, the dryness prevention device 1 can store a larger amount of liquid L in the liquid storage tank 11 and can suppress evaporation of water from the medium CM filled in the well 911 for a long period of time.

[0078] According to the drying prevention device 1 having the above-mentioned configuration, evaporation of water from the culture medium CM held in the well plate 90 can be prevented. Furthermore, the cell culture kit 100 configured as above includes the above-mentioned drying prevention device 1, and enables suitable cell culture.

[0079] [Second embodiment] 5 is an explanatory diagram of a drying prevention instrument 2A and a cell culture kit 200A according to a second embodiment of the present invention. In the following embodiments, the same reference numerals are used for the components already mentioned, and detailed description thereof will be omitted.

[0080] 5, a porous member 25 is provided at the steam port 10a. The porous member 25 is disposed so as to close the steam port 10a.

[0081] The porous member 25 has a function of using capillary action to soak up the liquid L as a whole member. The material forming the porous member 25 can be a sponge-like material having a plurality of pores that are internally connected to each other.

[0082] The material forming the porous member 25 may be an organic material such as a nonwoven fabric, a felt, a woven fabric, or a sponge, or an inorganic material such as a pumice stone or a porous ceramic. The sponge may be a natural sponge or a synthetic sponge.

[0083] The cell culture kit 200A includes the above-mentioned drying suppression device 2A and a well plate 90.

[0084] In the dryness suppression instrument 2A, the porous member 25 closes the steam port 10a, so that the liquid L is less likely to spill when the cell culture kit 200A is transferred, making it easy to handle.

[0085] Furthermore, when using the dryness prevention tool 2A, for example, the amount of liquid L may be adjusted so that the porous member 25 contacts the liquid L in the liquid storage tank 11 without the entire porous member 25 being submerged in the liquid L. The porous member 25 in contact with the liquid L absorbs the liquid L and promotes evaporation of the liquid L. As a result, in the cell culture kit 200A having the dryness prevention tool 2A, the peripheral portion of the well plate 90 (peripheral portion α of the region 12a) is filled with steam V during cell culture. As a result, in the cell culture kit 200A, evaporation of water from the medium CM in the well 911 is suppressed.

[0086] Alternatively, when the drying suppression device 2A is used, the porous member 25 may not contact the liquid L in the liquid storage tank 11, and the liquid level of the liquid L may be located below the porous member 25. Even in this case, the vapor V generated by evaporation of the liquid L is supplied to the peripheral portion α through the pores of the porous member 25 and fills the peripheral portion α. ​​As a result, in the cell culture kit 200A, evaporation of water from the medium CM in the well 911 is suppressed.

[0087] In addition to the above-mentioned effects, the drying suppression device 2A having the above-mentioned configuration can also suppress evaporation of water from the culture medium CM held in the well plate 90. Moreover, the cell culture kit 200A having the above-mentioned configuration also has the above-mentioned drying prevention device 1, and enables suitable cell culture.

[0088] (Modification) FIG. 6 is an explanatory diagram showing a dryness prevention tool 2B and a cell culture kit 200B according to a modified example of this embodiment, and corresponds to FIG.

[0089] 6, the porous member 26 of the dryness suppression device 2B is provided at the steam port 10a, and the lower end thereof contacts the bottom surface 11x of the liquid storage tank 11. As a result, the porous member 26 is always in contact with the liquid L regardless of the amount of the liquid L stored in the liquid storage tank 11, and the evaporation of the liquid L can be promoted.

[0090] 6, the porous member 26 may extend above the holding unit 12. The maximum amount of liquid L that can be stored in the liquid storage tank 11 is an amount at which the liquid level of the liquid L is at the same height as the upper end of the holding unit 12. Therefore, when the porous member 26 extends above the holding unit 12, the liquid L can be sucked up to a position higher than the liquid level of the liquid L in the absence of the porous member 26, and evaporation of the liquid L can be promoted.

[0091] In addition to the above-mentioned effects, the drying suppression device 2B configured as above can also suppress evaporation of water from the culture medium CM held in the well plate 90. Furthermore, the cell culture kit 200B configured as above also has the above-mentioned drying prevention device 2, and enables suitable cell culture.

[0092] [Third embodiment] FIG. 7 is an explanatory diagram of a dryness prevention tool 3A and a cell culture kit 300A according to a third embodiment of the present invention, and corresponds to FIG.

[0093] 7 includes a main body 30A and a lid 20. The main body 30A includes a liquid storage tank 31A, a holding portion 32A, and a support portion (not shown).

[0094] The liquid storage tank 31A has a bottom plate 311 and a side wall 312 extending upward from the periphery of the bottom plate 311. The bottom plate 311 has a light source 315 that irradiates ultraviolet light UV toward the inside of the liquid storage tank 31. The light source 315 is connected to an external power source 316.

[0095] A known light source capable of emitting ultraviolet light UV can be used as the light source 315. As the light source 315, for example, a light emitting diode (UV-LED) that emits ultraviolet light can be used.

[0096] The holder 32A has a bottom plate 321 and a side wall 322 extending upward from the periphery of the bottom plate 321. The bottom plate 321 has a light blocking means 325 at a position facing the light source 315.

[0097] The light shielding means 325 absorbs and attenuates ultraviolet rays UV. The light shielding means 325 is, for example, a black layer provided on the bottom surface 321x on the outside of the bottom plate 321. The light shielding means 325 is formed from a material having an absorption band for the wavelength of ultraviolet rays UV emitted from the light source 315. The light shielding means 325 may be formed by applying black paint, or may be formed by laminating a film-like or plate-like material on the bottom surface 321x. In addition, the function of the light shielding means 325 may be imparted to the entire holding portion 32A by forming the holding portion 32 using a resin material colored black.

[0098] Although the color of the light blocking means 325 is set to "black," the color is not limited to this. The color of the light blocking means 325 can be any of various colors that can attenuate ultraviolet light UV.

[0099] Further, a light shielding means 325 may be provided on either or both of the surface of the side wall 312 facing the liquid L stored in the liquid storage tank 31A and the surface of the side wall 322 of the holding part 32A facing the liquid L.

[0100] The cell culture kit 300A includes the above-mentioned drying suppression device 3A and a well plate 90.

[0101] In the dryness prevention device 3A configured as described above, the gap between the side wall 312 and the side wall 322 functions as the steam port 30a. In the dryness prevention device 3A, the steam V generated by evaporation of the liquid L is supplied to the peripheral portion α through the steam port 30a. As a result, in the well plate 90 placed in the dryness prevention device 3A, the moisture evaporated from the medium CM in the well 911 is less likely to diffuse toward the outside of the well plate 90, and drying of the medium CM is suppressed.

[0102] Furthermore, in the dryness suppression device 3A, ultraviolet rays UV are irradiated from the light source 315 to sterilize germs that may grow in the liquid L, thereby suppressing the growth of the germs. After passing through the liquid L, the ultraviolet rays UV reach the light blocking means 325 and are attenuated. Therefore, the ultraviolet rays UV do not reach the well plate 90, and do not inhibit the cell culture performed in the well plate 90.

[0103] The drying suppression device 3A having the above-mentioned configuration also makes it possible to suppress evaporation of water from the culture medium CM held in the well plate 90. Furthermore, the cell culture kit 300A having the above-mentioned configuration also has the above-mentioned drying suppression device 3A, and thus enables suitable cell culture.

[0104] (Modification) FIG. 8 is an explanatory diagram showing a dryness prevention tool 3B and a cell culture kit 300B according to a modified example of this embodiment, and corresponds to FIG.

[0105] 8, the drying prevention device 3B has a main body 30B and a lid 20. The main body 30B has a liquid storage tank 31B, a holding portion 32B, and a support portion (not shown).

[0106] The liquid storage tank 31B has a bottom plate 311 and a side wall 312. The side wall 312A has a light source 317 that irradiates ultraviolet light UV toward the inside of the liquid storage tank 31. The light source 317 is connected to an external power source 316. The light source 317 may be a light source having a similar configuration to the light source 315 described above.

[0107] Furthermore, liquid storage tank 31B has light shielding means 318 on side wall 312B opposite side wall 312A on which light source 317 is provided. Light shielding means 318 can be made of the same material as light shielding means 325 described above and can have the same properties.

[0108] In addition, a light shielding means 318 may be provided on either or both of the surface of the side wall 312 facing the liquid L stored in the liquid storage tank 31B and the surface of the side wall 322 of the holding portion 32B facing the liquid L.

[0109] The cell culture kit 300B includes the above-mentioned drying suppression device 3B and a well plate 90.

[0110] In the dryness suppression device 3B configured as described above, ultraviolet rays UV are irradiated from the light source 317 to sterilize germs that may grow in the liquid L, thereby suppressing the growth of the germs. After passing through the liquid L, the ultraviolet rays UV reach the light blocking means 318 and are attenuated. Therefore, the ultraviolet rays UV do not reach the well plate 90, and do not inhibit the cell culture performed in the well plate 90.

[0111] In addition to the above-mentioned effects, the drying suppression device 3B having the above-mentioned configuration can also suppress evaporation of water from the culture medium CM held in the well plate 90. Furthermore, the cell culture kit 300B configured as above also has the above-mentioned drying suppression device 3B, and enables suitable cell culture.

[0112] [Fourth embodiment] FIG. 9 is an explanatory diagram of a dryness prevention tool 4 and a cell culture kit 400 according to a fourth embodiment of the present invention, and corresponds to FIG.

[0113] The dryness prevention device 4 shown in Fig. 9 has a main body 40 and a lid 20. The main body 40 has a liquid storage tank 41, a holding portion 42, and a support portion (not shown).

[0114] The liquid storage tank 41 has a bottom plate 411 and a side wall 412 that is provided on the peripheral edge of the bottom plate 411 and extends upward. The bottom plate 411 has a light collecting window 415 that is made of an ultraviolet light transmitting material.

[0115] The light collecting window 415 transmits the ultraviolet light UV emitted from the external light source S and guides the light to the inside of the liquid storage tank 41. The light collecting window 415 is formed of a material that does not have an absorption band for the wavelength of the ultraviolet light UV. The light collecting window 415 may be formed of, for example, quartz glass.

[0116] The holder 42 has a bottom plate 421 and a side wall 422 extending upward from the periphery of the bottom plate 421. The bottom plate 421 has a light-shielding means 425 at a position facing the light-collecting window 415. The light-shielding means 425 can be made of the same material as the light-shielding means 325 described above, and can be configured to have the same properties.

[0117] In addition, a light shielding means 425 may be provided on either or both of the surface of the side wall 412 facing the liquid L stored in the liquid storage tank 41 and the surface of the side wall 422 of the holding part 42 facing the liquid L.

[0118] The cell culture kit 400 includes the above-mentioned drying suppression device 4 and a well plate 90.

[0119] In the dryness prevention device 4 configured as above, the gap between the side wall 412 and the side wall 422 functions as the vapor port 40a. In the dryness prevention device 3A, the vapor V generated by evaporation of the liquid L is supplied to the peripheral portion α through the vapor port 40a. As a result, in the well plate 90 placed in the dryness prevention device 4, the moisture evaporated from the medium CM in the well 911 is less likely to diffuse toward the outside of the well plate 90, and drying of the medium CM is suppressed.

[0120] Furthermore, in the dryness suppression device 4, the ultraviolet rays UV irradiated to the inside of the liquid storage tank 41 through the light collecting window 415 can sterilize germs that may grow in the liquid L and suppress the growth of the germs. After passing through the liquid L, the ultraviolet rays UV reach the light blocking means 425 and are attenuated. Therefore, the ultraviolet rays UV do not reach the well plate 90 and do not inhibit the cell culture performed in the well plate 90.

[0121] The drying suppression device 4 having the above-mentioned configuration can also suppress evaporation of water from the culture medium CM held in the well plate 90. Moreover, the cell culture kit 400 configured as above also has the above-mentioned drying prevention device 4, and enables suitable cell culture.

[0122] [Fifth embodiment] FIG. 10 is an explanatory diagram of a dryness prevention tool 5 and a cell culture kit 500 according to a fifth embodiment of the present invention, and corresponds to FIG.

[0123] 10 has a main body 50 and a lid 20. The main body 50 has a liquid storage tank 51 and a holder 52.

[0124] The liquid storage tank 51 has a bottom plate 511 and a side wall 512 that is provided on the peripheral portion of the bottom plate 511 and extends upward. A step that protrudes toward the inside of the liquid storage tank 51 is provided on the inner peripheral surface of the side wall 512. The step functions as a support portion in this embodiment. In the following description, the step provided on the side wall 512 is referred to as a support portion 53.

[0125] The support portion 53 may be provided on the inner circumferential surface of the side wall 512 in an endless closed ring shape in a plan view, or may be provided discretely at a plurality of positions on the inner circumferential surface.

[0126] Holder 52 is provided separably from storage tank 51, and covers an inner bottom surface 51x of storage tank 51. Holder 52 has a bottom plate 521 and a side wall 522 that is provided on the periphery of bottom plate 521 and extends upward. Side wall 522 surrounds an area 52a in holder 52 that holds well plate 90.

[0127] Holding portion 52 is supported by support portion 53 at the peripheral portion of outer bottom surface 521x of bottom plate 521. In the space surrounded by liquid storage tank 51 and holding portion 52, liquid L is stored.

[0128] A through hole penetrating the holding portion 52 in the thickness direction of the holding portion 52 is provided in the side wall 522 of the holding portion 52. The through hole functions as a steam port in this embodiment. In the following description, the through hole provided in the holding portion 52 is referred to as a steam port 50a. A lower end of the steam port 50a opens into a space in which the liquid L is stored.

[0129] The cell culture kit 500 includes the above-mentioned drying suppression device 5 and a well plate 90.

[0130] In the dryness suppression device 5 configured as described above, the vapor V generated by evaporation of the liquid L is supplied to the peripheral portion α through the vapor port 50a. As a result, in the well plate 90 placed in the dryness suppression device 5, the moisture evaporated from the medium CM in the well 911 is less likely to diffuse toward the outside of the well plate 90, and drying of the medium CM is suppressed.

[0131] In addition to the above-mentioned effects, the drying suppression device 5 having the above-mentioned configuration can also suppress evaporation of water from the culture medium CM held in the well plate 90. Moreover, the cell culture kit 500 configured as above also has the above-mentioned drying suppression device 5, and enables suitable cell culture.

[0132] [Sixth embodiment] FIG. 11 is an explanatory diagram of a dryness prevention tool 6 and a cell culture kit 600 according to a sixth embodiment of the present invention, and corresponds to FIG.

[0133] 11 includes a main body 60 and a lid 20. The main body 60 includes a liquid storage tank 61, a holding portion 62, and a support portion (not shown).

[0134] The liquid storage tank 61 has a bottom plate 611 and a side wall 612 extending upward from the periphery of the bottom plate 611. The bottom plate 611 has a heating member 615. The heating member 615 may be embedded in the bottom plate 611, or may be provided in contact with the surface of the bottom plate 611.

[0135] The retaining portion 62 may have a configuration similar to that of the retaining portion 12 described above.

[0136] The cell culture kit 600 includes the above-mentioned drying suppression device 6 and a well plate 90.

[0137] In the dryness prevention instrument 6, the heating member 615 indirectly heats the liquid storage tank 61, or directly heats the liquid L by contacting the liquid L. This makes it possible to heat the dryness prevention instrument 6 and keep the well plate 90 warm when the cell culture kit 600 is removed from the incubator.

[0138] Therefore, by using the dryness suppression device 6, even if there is a temperature difference between the temperature inside and outside the incubator, the effect of temperature changes on the cells cultured in the cell culture kit 600 can be reduced, and the cell culture conditions can be stabilized.

[0139] Furthermore, the gap between the side wall 612 and the holder 62 functions as a steam port 60a. In the dryness suppression device 3A, the steam V generated by evaporation of the liquid L is supplied to the peripheral portion α through the steam port 60a. As a result, in the well plate 90 placed in the dryness suppression device 6, the moisture evaporated from the medium CM in the well 911 is less likely to diffuse toward the outside of the well plate 90, and drying of the medium CM is suppressed.

[0140] The drying suppression device 6 having the above-mentioned configuration also makes it possible to suppress evaporation of water from the culture medium CM held in the well plate 90. Furthermore, the cell culture kit 600 configured as above also has the above-mentioned drying suppression device 6, and enables suitable cell culture.

[0141] Although the preferred embodiment of the present invention has been described above with reference to the accompanying drawings, the present invention is not limited to the above embodiment. The shapes and combinations of the components shown in the above embodiment are merely examples, and various modifications can be made based on design requirements, etc., without departing from the spirit of the present invention. EXAMPLES

[0142] The present invention will now be described in more detail with reference to examples, but the present invention is not limited to the following examples.

[0143] [Check for changes in liquid volume] Example 1 All wells of a 96-well plate conforming to the SBS standard were filled with 200 μL of water each, and then the well plate was covered. The liquid volume in the wells at the four corners of the well plate was measured, and the arithmetic mean value of the water volume in the wells at the four corners was calculated. The calculated arithmetic mean value was used as the initial value.

[0144] A cell culture kit having the same shape as the cell culture kit 100 shown in Figures 1 and 2 was prepared, and pure water was stored in the liquid storage tank. The well plate with the lid was placed on the holder of the cell culture kit, and after the lid was closed, it was kept in an incubator at a temperature of 37°C and a humidity of 90% or more.

[0145] After a predetermined time had elapsed, the liquid volumes in the wells at the four corners of the well plate were measured, and the arithmetic mean values ​​of the liquid volumes in the wells at the four corners were calculated. Furthermore, the rate of change in the liquid volume relative to the initial value was calculated for the calculated arithmetic mean values.

[0146] The amount of liquid in the wells was measured using a plate reader. A calibration curve was prepared in advance based on the relationship between the absorbance of a 970 nm laser beam and the amount of liquid in the wells, and the amount of liquid in each well was calculated from the absorbance of each well measured using the plate reader.

[0147] Comparative Example 1 The rate of change in the liquid volume in the well was determined in the same manner as in Example 1, except that the cell culture kit was not used.

[0148] Fig. 12 is a graph showing the rate of change in the liquid volume in Example 1 and Comparative Example 1. As shown in Fig. 12, it was found that the use of the cell culture kit can prevent the drying of the four corners of the well plate. From this result, it was found that the use of the cell culture kit can be expected to prevent the drying of the outer periphery of the well plate.

[0149] [Confirmation of effects on cell culture] Example 2 Neurons were seeded in all wells of a 48-well MEA (Micro Electrode Array) plate manufactured by Axion. Neurons were differentiated from iPS-derived healthy individual lines. The neurons were mixed with primary astrocytes and seeded in the wells. Human Primary Astrocytes (Thermo Fisher Scientific) were used as astrocytes.

[0150] A cell culture kit having the same shape as the cell culture kit 100 shown in Figures 1 and 2 was prepared, and pure water was stored in the storage tank. The well plate with the lid was placed on the holder of the cell culture kit, and after the lid was closed, the nerve cells were cultured in an incubator at a temperature of 37°C and a humidity of 90% or more.

[0151] After culturing the neurons for 4 weeks, the action potential of the neurons was measured using Meastro (Axion Corp.) The number of spontaneous spikes was measured for 10 minutes in all wells, and the variance of the number of spontaneous spikes per well was calculated.

[0152] The spontaneous firing rate of the cultured neurons was then measured for 10 minutes after the addition of a drug, a dimethyl sulfoxide (DMSO) solution of picrotoxin, a GABA antagonist.

[0153] First, 300 μL of 1% by volume DMSO aqueous solution was added to each of the wells in one row (6 wells), and the number of spontaneous firings was measured for 10 minutes.

[0154] Next, picrotoxin DMSO solution was gradually added to the same well until the picrotoxin concentration in the well became 0.1 μmol / L, 1 μmol / L, and 10 μmol / L, and the number of spontaneous firings was measured for 10 minutes at each picrotoxin concentration. The variance of the number of spontaneous firings for 10 minutes was calculated for each picrotoxin concentration.

[0155] Comparative Example 2 The variation in the number of spontaneous firings of neurons per well was determined in the same manner as in Example 2, except that the cell culture kit was not used.

[0156] Fig. 13 is a diagram showing the variation in the number of spontaneous firings of neurons per well for the cultured neurons in Example 2 and Comparative Example 2. As shown in Fig. 13, Example 2, in which a cell culture kit was used, showed smaller variation than Comparative Example 2, indicating that the use of a cell culture kit suppresses variation in maturation of cultured cells.

[0157] Fig. 14 is a diagram showing the response variability of neurons to drugs after culture in Example 2 and Comparative Example 2. As shown in Fig. 13, Example 2, in which a cell culture kit was used, showed smaller response variability than Comparative Example 2, indicating that the use of a cell culture kit can suppress the response variability of cultured cells.

[0158] [Checking the rate of carbon dioxide inflow] Example 3 The dryness suppression device shown in Fig. 1 was used. The size of the ventilation hole 20a in a plan view was 79 mm x 106 mm. The size of the area 12a in a plan view was 82 mm x 124 mm. The end of the lid 20 of the dryness suppression device used on the ventilation hole side overlapped with the area 12a in a plan view.

[0159] An NDIR (non-dispersive infrared) CO2 concentration sensor (HJ-CO2-LOG, manufactured by Sato Shoji Co., Ltd.) was placed in the holder of the dryness suppression device, and the dryness suppression device was placed in an atmosphere with a CO2 concentration of 5% and a humidity of 90% (relative humidity, @37°C). The "rate of increase in carbon dioxide concentration per unit time" obtained from the carbon dioxide measurement value by the CO2 concentration sensor was calculated as the "rate of carbon dioxide inflow."

[0160] Comparative Example 3 The "carbon dioxide inflow rate" was determined in the same manner as in Example 3, except that a commercially available drying prevention device (Olaf evaporation prevention chamber, manufactured by Nippon Genetics Co., Ltd.) was used.

[0161] The commercially available product used is designed to seal the space in which the well plate is placed in order to prevent the water in the well plate from evaporating. However, the commercially available product used does not have a liquid storage tank for storing a liquid for preventing drying, and does not meet the requirements of the present invention.

[0162] The commercially available product used has a ventilation hole in the side wall of the main body that houses the well plate, and a filter is provided to cover the ventilation hole.

[0163] The measurement results are shown in Figure 15. As a result of the measurement, the inflow rate of carbon dioxide into the dryness suppression device used in Example 3 was 180 ppm / sec. On the other hand, the inflow rate of carbon dioxide into the commercially available product used in Comparative Example 3 was 4 ppm / sec.

[0164] When the dryness prevention device of the present invention (Example 3) is used, the inflow rate of carbon dioxide becomes 20 ppm / sec or more, and it is easy to uniformly control the culture conditions inside the dryness prevention device.

[0165] On the other hand, the desiccation prevention device of Comparative Example 3 was able to prevent desiccation by sealing the well plate, but it was found that the inflow rate of carbon dioxide was low and it was difficult to uniformly control the culture conditions inside the desiccation prevention device.

[0166] The present invention includes the following aspects.

[0167] [1] A dryness suppression device comprising a main body for accommodating a well plate and a lid provided on the main body in a freely opening and closing manner, the main body having a holding portion for holding the well plate, a liquid storage tank for storing liquid, and a support portion for supporting and spacing the holding portion from the inner bottom surface of the liquid storage tank, wherein when the lid is closed, the internal space enclosed by the liquid storage tank and the lid is connected to the outside.

[0168] [2] The drying suppression device described in [1], wherein at least one of the lid or the liquid storage tank has a ventilation hole connecting the internal space with the outside.

[0169] [3] The drying suppression device described in [2], wherein the liquid storage tank has an opening that opens above the holding portion, the lid covers the opening, and the ventilation hole is provided in the lid.

[0170] [4] A dryness suppression device as described in [3], wherein, in a plan view, the holding portion has a steam port around it through which steam of the liquid stored in the liquid storage tank can flow, the lid overlaps with the steam port in a plan view, and the ventilation hole overlaps with an area of ​​the holding portion that holds the well plate in a plan view, but does not overlap with the steam port in a plan view.

[0171] [5] A drying suppression device described in any one of [1] to [4], wherein the holding portion has a bottom plate on which the well plate is placed and a side wall surrounding an area of ​​the bottom plate that holds the well plate.

[0172] [6] A dryness suppression device as described in any one of [1] to [5], wherein, in a plan view, the periphery of the holding portion has a steam port through which steam of the liquid stored in the liquid storage tank can flow, and the steam port is provided with a porous material.

[0173] [7] The drying suppression device according to [6], wherein the porous member blocks the steam port.

[0174] [8] A dryness suppression device as described in [6] or [7], wherein the porous member extends upward beyond the holding portion.

[0175] [9] A drying suppression device described in any one of [6] to [8], wherein the porous member extends to the bottom surface.

[0176]

[10] The dryness suppression device described in any one of [1] to [9], wherein the main body portion has a light source that emits ultraviolet light toward the inside and a shading means that is arranged opposite the light source.

[0177]

[11] The drying suppression device described in

[10] , wherein the light source is provided on a bottom plate of the liquid storage tank, and the light-shielding means is provided on an outer bottom surface of the holding portion.

[0178]

[12] A dryness suppression device as described in

[10] or

[11] , wherein the light source is provided on a side wall of the liquid storage tank, and the shading means is provided on a side wall opposite to the side wall on which the light source is provided.

[0179]

[13] A drying suppression device described in any one of [1] to [9], wherein the bottom plate of the liquid storage tank has a light-gathering window formed of an ultraviolet light-transmitting material, and the holding portion has a light-shielding means at a position opposite the light-gathering window.

[0180]

[14] A dryness suppression device as described in any one of [1] to

[13] , wherein the main body portion is such that the liquid storage tank and the support portion are separable from the holding portion.

[0181]

[15] A dryness suppression device as described in any one of [1] to

[14] , wherein, in a plan view, the periphery of the holding portion has a steam port through which steam of the liquid stored in the liquid storage tank can flow, the holding portion is spaced apart from a side wall of the liquid storage tank in a plan view, and the gap between the holding portion and the side wall is the steam port.

[0182]

[16] A dryness suppression device as described in any one of [1] to

[15] , wherein, in a plan view, the periphery of the holding part has a steam port through which steam of the liquid stored in the liquid storage tank can flow, the steam port being a through hole penetrating the holding part in the thickness direction of the holding part, and the through hole is provided outside the area of ​​the holding part that holds the well plate.

[0183]

[17] A drying suppression device according to any one of [1] to

[16] , wherein the liquid storage tank has a heating member.

[0184]

[18] A dryness suppression device as described in any one of [1] to

[17] , wherein in an atmosphere with a CO2 concentration of 5%, the inflow rate of CO2 from the atmosphere to the internal space is 20 ppm / sec or more.

[0185]

[19] A drying suppression device comprising: a main body for accommodating a well plate; and a lid provided on the main body that can be opened and closed freely, the main body having a holding part for holding the well plate, a liquid storage tank for storing liquid, and a support part for supporting the holding part at a distance from the inner bottom surface of the liquid storage tank, wherein the remaining water rate of the liquid in the wells at the four corners of a well plate placed on the holding part is 75% or more as determined by the measurement method described below. (Measurement method) A 96-well plate conforming to SBS standards, in which all wells are filled with 200 μL of water, is placed in the main body, water is stored in the storage tank, and the plate is left with the lid on. After leaving the plate for 20 days, the rate of change in the amount of water relative to the initial value is calculated based on the following formula (1). (Water remaining rate)(%)=B / A×100…(1) (A indicates the average initial amount of water in the four wells located at the four corners of the well plate. B indicates the average amount of water remaining in the four wells located at the four corners of the well plate.)

[0186]

[20] A cell culture kit comprising: a drying suppression device described in any one of [1] to

[19] ; ​​a well plate housed in the drying suppression device and having a plurality of wells; cells housed inside the wells; and culture medium filled inside the wells. [Explanation of symbols]

[0187] 1, 1 , 11x,51x,321x,521x...bottom, 12,32,32A,32B,42,52,62...holding part, 12a,52a...area, 13,53...support part, 20,20X,92...lid, 20a...ventilation hole, 22,112,122,312,312A,312B ,322,412,422,512,522,612,922...side wall, 25,26...porous member, V,V1...steam, 90...well plate, 100,100X,200A,200B,300A,300B,400,500,600...cell culture kit, 111,121,311,321,411,421,511,521,611...bottom plate, 315,317...light source, 318,325,425...light shielding means, 415...light window, 615...heating member, 911...well, Air...air, CM...culture medium, L...liquid, UV...ultraviolet light [Prior art documents] [Patent documents]

[0188] [Patent Document 1] JP 2000-236869 A

Claims

1. A main body that accommodates a well plate; A lid provided on the main body in an openable and closable manner, The main body portion includes a holder for holding the well plate, A liquid storage tank for storing liquid; a support portion that supports the holding portion at a distance from a bottom surface of an inner side of the liquid storage tank, When the lid is closed, an internal space surrounded by the liquid storage tank and the lid is in communication with the outside, At least one of the lid and the liquid storage tank has a ventilation hole connecting the internal space with the outside, The liquid storage tank has an opening that opens above the holding portion, The lid covers the opening, The ventilation hole is provided in the lid, a vapor port through which vapor of the liquid stored in the liquid storage tank can flow, the vapor port being provided around the holding portion in a plan view; The lid overlaps with the steam vent in a plan view, The ventilation hole overlaps with an area of ​​the holding portion that holds the well plate in a plan view, and does not overlap with the steam vent in a plan view.

2. The holder includes a bottom plate on which the well plate is placed, The drying suppression device according to claim 1 , further comprising: a side wall provided to surround an area of ​​the bottom plate that holds the well plate.

3. a vapor port through which vapor of the liquid stored in the liquid storage tank can flow, the vapor port being provided around the holding portion in a plan view; 3. The dryness suppression device according to claim 1, wherein the steam port is provided with a porous member.

4. The drying suppression device according to claim 3 , wherein the porous member closes the steam port.

5. 5. The dryness prevention device according to claim 3, wherein the porous member extends above the holding portion.

6. The drying suppression device according to claim 3 , wherein the porous member extends to the bottom surface.

7. The main body portion includes a light source that emits ultraviolet light toward the inside, The dryness suppression tool according to claim 1 , further comprising a light blocking means provided opposite to the light source.

8. The light source is provided on a bottom plate of the liquid storage tank, 8. The dryness suppression device according to claim 7, wherein the light blocking means is provided on an outer bottom surface of the holding portion.

9. The light source is provided on a side wall of the liquid storage tank, 9. The dryness suppression device according to claim 7, wherein the light blocking means is provided on a side wall opposite to the side wall on which the light source is provided.

10. the bottom plate of the liquid storage tank has a light-collecting window made of an ultraviolet light-transmitting material; The dryness prevention tool according to claim 1 , wherein the holding portion has a light-shielding means at a position opposite to the light-collecting window.

11. The dryness prevention device according to claim 1 , wherein the main body portion is configured such that the liquid storage tank and the support portion are separable from the holding portion.

12. a vapor port through which vapor of the liquid stored in the liquid storage tank can flow, the vapor port being provided around the holding portion in a plan view; The holding portion is spaced apart from a side wall of the liquid storage tank in a plan view, The dryness suppression device according to claim 1 , wherein a gap between the holding portion and the side wall is the steam vent.

13. a vapor port through which vapor of the liquid stored in the liquid storage tank can flow, the vapor port being provided around the holding portion in a plan view; the steam port is a through hole penetrating the holding portion in a thickness direction of the holding portion, The drying prevention device according to claim 1 , wherein the through-hole is provided outside a region of the holding portion that holds the well plate.

14. The dryness suppression device according to claim 1 , wherein the liquid storage tank has a heating member.

15. CO ２ In an atmosphere with a concentration of 5%, ２ 15. The dryness suppression device according to claim 1, wherein the inflow rate of the dryness suppression device is 20 ppm / sec or more.

16. A dryness suppression device described in any one of claims 1 to 15, wherein the remaining water rate, as determined by the measurement method described below, for the liquid volume in the wells at the four corners of the well plate placed on the holding portion is 75% or more. (Measurement method) An SBS standard 96-well plate in which all wells are filled with 200 μL of water is placed in the main body, water is stored in the storage tank, and the plate is left in the state in which the lid is closed. After leaving it for 20 days, the rate of change in the amount of water relative to the initial value is calculated based on the following formula (1). (Water Residual Rate) (%) = B / A × 100 ... (1) (A indicates the average initial water volume in a total of four wells located at the four corners of the well plate. B indicates the average amount of water remaining in a total of four wells located at the four corners of the well plate. ）

17. A dryness suppression device according to any one of claims 1 to 16, A well plate housed in the drying suppression device and having a plurality of wells; A cell contained within the well; A cell culture kit having a culture medium filled inside the well.

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