JP2023175345A - Cell container and method for use thereof - Google Patents

Abstract

To provide a cell container that reduces the mixing of impurities during cell aspiration.SOLUTION: A cell container has a recess for accommodating cells. On the bottom of the recess, disposed are a plurality of structures that have a specific gravity of greater than 1, show water-insolubility and bio-compatibility, and are composed of resin or metal.SELECTED DRAWING: Figure 1

Description

The present invention relates to a cell storage container and a method of using the same.

Containers for culturing cells include containers for culturing adherent cells that have a surface treatment that makes it easier for cells to adhere to the container surface, and containers for floating culture that have a surface treatment that makes it difficult for cells to adhere to the container surface. In some cases, they are processed to suit various purposes, such as containers for culturing (see Patent Document 1).

When culturing cells, the cell suspension or culture solution added to the culture container may contain unnecessary substances such as impurities derived from dead cells and exosomes generated by cell metabolism. There is. Additionally, dust, lint, and the like derived from the experimental environment may be mixed in, but these impurities are unnecessary for maintaining a good cell condition, and it is desirable to be able to remove them. However, these substances are minute and difficult to remove using a micropipette or the like.

Therefore, containers having filters and micropores for removing impurities in cell culture fluids and cell suspensions have been developed (see Patent Documents 2 to 4). However, in the process of removing contaminants using filters and micropores, the culture medium is also absorbed at the same time as the contaminants are collected, resulting in a decrease in the culture medium and the environment around the cells being cultured. It will change.

On the other hand, a cell mass formed by a plurality of cells adhering to each other requires seeding a large number of cells in one culture compartment. Therefore, the frequency of contamination with impurities such as dead cells increases. On the other hand, it is difficult to remove only impurities such as dead cells during culture. However, unnecessary impurities may inhibit adhesion between cells and metabolism, so it is desirable to remove them. Furthermore, when transplanting a cell mass into a living body, it is desirable to reduce the introduction of contaminants as much as possible, but when the cell mass is aspirated with a micropipette, contaminants contained in the culture medium are also aspirated at the same time. In many cases, it is difficult to remove by suction alone.

When transplanting a cell mass or the like into a living body, it is necessary to replace the medium with a buffer solution in order to remove the medium and impurities contained in the medium. Although it is possible to remove contaminants by replacing the liquid, it is difficult to remove contaminants that have adhered to the culture container. Therefore, it is effective to aspirate the cell mass and transfer it to a cell storage container different from the culture container. In this case, it is necessary to remove contaminants brought in when the cell mass is sucked from the culture container.

International Publication No. 2021/167042 Japanese Patent Application Publication No. 2006-94718 International Publication No. 2020/054755 JP 2021-112142 Publication

The present invention has been made in view of these problems, and an object of the present invention is to provide a cell storage container in which contaminants are less likely to be mixed in when cells are aspirated.

A cell storage container for solving the above problems is a cell storage container having a recess for storing cells, and is made of resin or metal, which has a specific gravity greater than 1, is water-insoluble, and is biocompatible. A plurality of structures are disposed on the bottom surface of the recess.

According to the above-mentioned cell storage container, since the structure is placed on the bottom of the culture container, when cells are placed in the storage container, a gap is formed between the cells and the bottom of the cell storage container, and the surroundings of the culture medium etc. Contaminants with a higher specific gravity than the liquid will settle. In addition, since it does not elute into the culture medium, it does not have a large effect on the environment around cells. Furthermore, structures made of biocompatible materials are not toxic to living organisms, so when the structures are unintentionally inserted into living organisms, they do not cause foreign body reactions or rejection reactions. It becomes possible to ensure safety. Furthermore, resins and metals are easy to mold, making it possible to create structures of arbitrary shapes. This separates cells and impurities, making it possible to reduce the amount of impurities mixed in when aspirating cells.

Further, the structure may be made of one or more materials selected from the group consisting of polylactide (PLA), polycaprolactone (PCL), polyglycolide (PGA), and polyesters based on copolymers thereof, or a plurality of materials. It may also be constructed from a combination of materials.

According to the above configuration, when the structure is unintentionally inserted into a living body, a foreign body reaction or a rejection reaction does not occur, and safety can be ensured.

The arithmetic mean roughness (Ra) of the structure may be 0.03 μm or less.

According to the above configuration, the physical stimulation exerted on the cell membrane by the surface of the structure is reduced, making it possible to suppress unintended damage to the cells.

The surface of the structure may be treated so that it does not adhere to cells.

According to the above configuration, it is possible to suppress the frequency at which the structure adheres to cells, and it is possible to prevent unintentional contamination of the structure when cells are aspirated and used for another purpose. Become.

The shape of the structure may be cylindrical.

According to the above configuration, the structure has few corners, and physical damage to cells is reduced, making it possible to suppress a decrease in cell activity.

The maximum length of a straight line connecting the ends of the structure may be equal to or longer than the diameter of the suction port of the device for aspirating cells.

According to the above configuration, it is possible to suppress the frequency of structures being sucked into the device when cells are sucked, and structures may be unintentionally mixed in when cells are sucked and used for another purpose. It becomes possible to prevent this.

The structure may have a bent portion.

According to the above configuration, since the voids formed between the structures become larger, it becomes possible to trap large contaminants such as dead cells.

The shape of the structure may have a branched portion.

According to the above configuration, since the voids formed between the structures become larger, it becomes possible to trap large contaminants such as dead cells.

A method of using a cell storage container to solve the above problem is to discharge cells into a recess together with a liquid, place the cells on a structure placed on the bottom of the recess, and flatten the water surface of the liquid. After allowing enough time for contaminants to precipitate, aspirate the cells.

Further, the cell storage container has a structure made of one or more materials selected from the group consisting of polylactide (PLA), polycaprolactone (PCL), polyglycolide (PGA), and polyester based on copolymers thereof. Alternatively, it may be made of a combination of a plurality of materials, have an arithmetic mean roughness (Ra) of 0.03 μm or less, and be surface-treated to be non-adhesive to cells.

According to the present invention, it is possible to provide a cell storage container in which contaminants are less likely to be mixed in when cells are aspirated.

FIG. 1 is a cross-sectional view showing a cell storage container according to an embodiment. A sectional view showing an example of a structure of a cell storage container in an embodiment. A sectional view showing an example of a structure of a cell storage container in an embodiment. A sectional view showing an example of a structure of a cell storage container in an embodiment. A sectional view showing an example of a structure of a cell storage container in an embodiment. A sectional view showing an example of a structure of a cell storage container in an embodiment. FIG. 3 is a cross-sectional view showing how to use the cell storage container in the embodiment. FIG. 3 is a cross-sectional view showing how to use the cell storage container in the embodiment. FIG. 3 is a cross-sectional view showing how to use the cell storage container in the embodiment. FIG. 3 is a cross-sectional view showing how to use the cell storage container in the embodiment. FIG. 3 is a cross-sectional view showing how to use the cell storage container in the embodiment. FIG. 2 is a cross-sectional view showing an example of a method for manufacturing a cell storage container. FIG. 3 is a cross-sectional view showing a modified example of the cell storage container. FIG. 3 is a cross-sectional view showing a modified example of the cell storage container. FIG. 3 is a cross-sectional view showing a modified example of the cell storage container. FIG. 3 is a cross-sectional view showing a modified example of the cell storage container. FIG. 3 is a cross-sectional view showing a modified example of the cell storage container. FIG. 3 is a cross-sectional view showing a modified example of the cell storage container. FIG. 3 is a cross-sectional view showing a modified example of the cell storage container. FIG. 3 is a cross-sectional view showing a modified example of the cell storage container. FIG. 3 is a cross-sectional view showing a modified example of the cell storage container. FIG. 3 is a cross-sectional view showing a modified example of the cell storage container. FIG. 3 is a cross-sectional view showing a modified example of the cell storage container.

(Embodiment)
An embodiment of a cell storage container will be described with reference to FIGS. 1 to 6. FIG. 1 is a cross-sectional view showing a cell storage container according to an embodiment, and FIGS. 2 to 6 are cross-sectional views showing an example of the structure of the cell storage container in the embodiment.

The cell storage container 100 includes a recess 2 and a structure 1. Although details will be described later, the cell group 104 and the liquid material 101 are accommodated in the recess 2, and the cell group 104 in the recess 2 is aspirated with an instrument 6 such as a micropipette. The recess 2 has a U-shaped, V-shaped, or U-shaped bottom surface. The structure 1 extends from the support surface 20 of the recess 2. The cell storage container 100 and the structure 1 are made of resin or metal, and may be joined with an adhesive or by a joining method such as thermal welding or welding by applying ultrasonic vibration. Furthermore, the cell container 100 and the structure 1 may be made of the same material or different materials. For example, if the cell storage container 100 is made of PDMS (polydimethylsiloxane) and the structure 1 is made of PEEK (polyetheretherketone), it is possible to reduce the possibility of having an unfavorable effect on the cells. becomes. In other words, when cells are housed together with a liquid, it is possible to maintain a good environment around the cells in a cell container made of PDMS due to its high oxygen permeability, and if the structure 1 is made of PEEK, it is biocompatible. It exhibits high chemical resistance, high chemical resistance, and low elution properties, so it does not have a large effect on the environment around cells. The central axis of the structure 1 may or may not be perpendicular to the lower surface (upper surface) of the cell storage container 100. That is, the structure 1 may extend obliquely from the support surface 20 (FIG. 2). The maximum length l of a straight line connecting both ends of the structure 1 is preferably 4/5 or less of the depth h of the recess 2, and preferably 1/2 or less of the inner diameter d of the recess 2 ( Figure 1). Note that the length l indicates the length in the major axis direction if the structure 1 is columnar.

As shown in FIG. 2, when a plurality of structures 1 extend from the support surface 20 of the recess 2, all the structures 1 may be oriented in the same direction, and one of the plurality of structures 1 may be oriented in the same direction. Either one may extend in different directions.

As shown in FIG. 3, when a plurality of structures 1 extend from the support surface 20 of the recess 2, the ends of the structures 1 that are not in contact with the support surface 20 may not all be on the same plane. . That is, the lengths of the structures in the recess 2 may be the same or different. As shown in FIG. 1, when the ends of the structure 1 that are not in contact with the support surface 20 are all on the same plane, when aspirating cells, the distance between the tip of the suction device and the structure is Since it becomes easy to predict, it becomes possible to reduce the probability of cells being crushed between the structure 1 and the device. On the other hand, as shown in FIG. 3, if the ends of the structure 1 that are not in contact with the support surface 20 are not on the same plane, for example, when housing a dumbbell-shaped cell mass, the ends of the structure 1 By arranging the cell clusters in a dumbbell shape that matches the three-dimensional structure of the cell clusters, it becomes possible to arrange the cell clusters in any direction.

As shown in FIG. 4, the structure 1 does not need to be adhered to the bottom surface of the recess 2 of the cell container 100, and may be placed in the recess 2 in a precipitated state. The volume ratio between the structure 1 and the recess 2 may be smaller than 1:1, and preferably the volume ratio between the structure 1 and the recess 2 is 1:2. Note that when a plurality of structures 1 are deposited and arranged on the bottom surface of the recess 2, the sizes and shapes of the structures 1 may all be the same or different.

The structure 1 has a specific gravity greater than 1 and is made of resin or metal. Specifically, polycaprolactone, L-lactic acid caprolactone copolymer, polydioxanone, glycolide lactic acid copolymer, polyDL lactic acid, polycarbonate, polyether ether ketone, engineering plastic, ultra-high molecular weight polyethylene, fluororesin, polylactic acid, titanium, It is composed of one or more materials selected from the group consisting of stainless steel, platinum, Co--Cr alloy, alumina, and zirconia ceramics, or a combination of multiple materials. Furthermore, since the structure 1 is water-insoluble, it does not elute into the culture medium or the like, so it does not have a large effect on the environment around the cells. This makes it difficult to cause unintended damage to cells. Furthermore, since the structure 1 made of biocompatible material is not toxic to living organisms, it will not cause foreign body reactions or rejection reactions when the structure 1 is unintentionally inserted into the living body. This makes it possible to ensure safety. In addition, resin and metal can be easily molded, making it possible to create a structure 1 of any shape. More preferably, one or more materials selected from the group consisting of polylactide (PLA), polycaprolactone (PCL), polyglycolide (PGA), and polyesters based on copolymers thereof, or a plurality of materials. It is better to consist of combinations. These materials are resins and can be easily molded, making it easy to mold structures having arbitrary shapes. Furthermore, since it exhibits biocompatibility and biodegradability, it does not exhibit toxicity in vivo, improving safety.

It is desirable that the arithmetic mean roughness (Ra) of the structure 1 is 0.03 μm or less. This reduces the physical stimulation that the surface of the structure 1 gives to the cell membrane, making it possible to suppress unintended damage to the cells.

The structure 1 may be subjected to a surface treatment that makes it non-adhesive to cells, such as coating with a surfactant such as Pluronic (registered trademark). This makes it possible to suppress the frequency at which Structure 1 adheres to cells, and prevents Structure 1 from being unintentionally mixed in when cells are aspirated and used for another purpose. .

As shown in FIG. 5, the cell group 104 is accommodated in the recess 2. Further, a liquid body 101 such as a cell culture solution or physiological saline may be contained at the same time. If the structure 1 is not adhered to the support surface 20, the structure 1 is placed on the bottom surface of the recess 2 before the cell group 104 is placed, and then the cell group 104 is placed in the recess 2. Note that what is housed in the recess 2 is not limited to the cell group 104, but may be an effective substance such as a drug, a transplant, or a cell.

As shown in FIG. 6, the cell group 104 accommodated in the recess 2 may be aspirated with an instrument 6 such as a micropipette. For example, when the recess 2 is filled with the liquid 101, a plurality of metal structures 1 are precipitated in the recess 2, and at the same time, the cell group 104 is placed on the precipitated structures 1. When suctioning the cell group 104, the suction port of the instrument 6 is placed very close to the cell group 104, and when negative pressure is generated, the cell group 104 is attracted to the suction port, but the structure 1 It is desirable that it not be sucked into the device 6.

Cell group 104 includes at least a cell group to be transplanted into a living body. The puncture target to which the cell group 104 is transplanted may be, for example, at least one of intradermal and subcutaneous. Further, the object to be punctured may be inside a tissue such as an organ.

The cell group 104 may be an aggregate of a plurality of aggregated cells, an aggregate of a plurality of cells connected by intercellular bonds, or may be composed of a plurality of dispersed cells. The cells constituting the cell group 104 may be undifferentiated cells, cells that have completed differentiation, or may include both undifferentiated cells and differentiated cells. The cell group 104 is, for example, a cell mass such as a spheroid, an primordium, a tissue, an organ, an organoid, a mini-organ, or the like.

The cell group 104 has the ability to affect tissue formation in a living body by being placed in a puncture target. An example of the cell group 104 is a cell aggregate containing cells having stem cell properties. An example of the cell group 104 contributes to hair growth or growth by being placed intradermally or subcutaneously.

These cell groups 104 have the ability to function as a hair follicle primordium, the ability to differentiate into hair follicle organs, the ability to induce or promote the formation of hair follicle organs, the ability to induce or promote hair formation in hair follicles, etc. have Further, the cell group 104 may include cells that contribute to hair color control, such as stem cells that differentiate into pigment cells, or may include vascular cells.

The cell group 104 in this embodiment is a primitive organ primordium. Organ primordia contain mesenchymal cells and epithelial cells. Organ primordia include hair follicle primordia that differentiate into hair follicle organs, liver primordium, kidney primordium, pancreatic primordium, nervous system primordium, vascular system primordium, and the like.

Hair follicle primordia are produced by culturing a mixture of mesenchymal cells derived from mesenchymal tissues such as dermal papillae and epithelial cells derived from epithelial tissues located in the bulge region, hair bulb base, etc. under predetermined conditions. It can be formed by However, the method for producing hair follicle primordia is not limited to the above example. The origin of the mesenchymal cells and epithelial cells used to produce the hair follicle primordium is also not limited, and these cells may be cells derived from the hair follicle organ or cells different from the hair follicle organ. The cells may be derived from an organ or may be cells derived from pluripotent stem cells.

Note that the cell group 104 may include cells other than cells that contribute to hair growth or hair growth. The volume of the cell group 104 only needs to be smaller than the volume of the recess 2.

When the liquid body 101 is accommodated together with the cell group 104, nutrients can be supplied to the cell group 104, which has the effect of suppressing a decrease in the activity of transplants such as cells. By retaining the cell group 104 in the liquid body 101, damage to the cell group 104 due to contact or friction between the cell group 104 and the structure 1 can be reduced. It is desirable that the liquid 101 is a liquid that has little effect on the living body when injected into the living body. For example, the liquid 101 is a liquid that protects the skin, such as physiological saline, petrolatum, or lotion, or a mixture of these liquids. The liquid body 101 may be a medium for cell culture, or may be a liquid replaced from the medium. The liquid body 101 may contain additional components such as nutritional components, and may also contain components necessary for the survival of cells such as oxygen. Furthermore, the liquid body 101 may be a low viscosity fluid or a high viscosity fluid, or may be a sol-like body or a gel-like body.

Next, a method of using the cell container 100 will be described with reference to FIGS. 7 to 11. 7 to 11 are cross-sectional views showing how to use the cell storage container in the embodiment.

As shown in FIG. 7, the structure 1 may be adhered to the support surface 20 of the recess 2 of the cell storage container 100, or may be arranged by precipitation.

As shown in FIG. 8, a cell group 104, which is a transplant, is held together with a liquid material 101 in a tray 5 such as a culture container. For example, the tray 5 has a culture recess 50 in which the liquid material 101 and the cell group 104 are accommodated. The tray 5 is not limited to the one illustrated in FIG. 7, and the cell group 104 and the liquid 101 may be arranged on a flat surface of the tray 5. When the tray 5 is a culture container and the cell group 104 is cultured in the tray 5, the liquid body 101 may be a medium for culturing the cells or a liquid replaced from the medium. Good too. When the cell group 104 is cultured in the tray 5, impurities 105 such as dead cells that do not aggregate with other cells and fine lint that is unintentionally mixed in during the operation step are collected in the liquid body 101. It may be mixed. This contaminant 105 may be floating in the liquid body 101 or may be caught on unevenness on the surface of the cell group 104.

As shown in FIG. 9, the cell group 104 cultured in the culture recess 50 of the tray 5 is aspirated using an instrument 6 such as a micropipette.

As shown in FIG. 10, the cell group 104 sucked by the device 6 is discharged into the recess 2 of the cell storage container 100. At this time, not only the cell group 104 but also the liquid 101 may enter the recess 2 . The volume of the liquid body 101 may be equal to or less than the volume of the recess 2. Preferably, the interface between the liquid body 101 and air is located at a higher position than the upper end of the cell group 104, and the cell group 104 Good condition inside.

When the cell group 104 is placed in the recess 2 and a period of time long enough for the water surface of the liquid 101 to become flat and for the contaminants 105 to precipitate, the contaminants 105 contained in the liquid 101 will precipitate on the bottom surface of the recess 2. . At this time, the interval between adjacent structures 1 is configured to be smaller than the cell group 104 and larger than the impurities 105. Therefore, although the cell group 104 is supported by the upper ends of the plurality of structures 1, the contaminants 105 are not supported by the upper ends of the structures 1 and precipitate in the gaps formed between adjacent structures 1. For example, if it is assumed that the impurities 105 are smaller than 100 μm, the interval (gap) between adjacent structures 1 may be 100 μm or more. Further, when it is assumed that a normal cell group 104 has a diameter of 500 μm or more, the interval between adjacent structures 1 may be less than 500 μm. Specifically, assuming that those with a diameter of 500 μm or more are judged as normal cell groups 104 and those with a diameter of 300 μm or less as contaminants 105, by arranging the structures 1 with a spacing of 400 μm, normal cell groups 104 are determined. Physical separation of the cell group 104 and the contaminants 105 becomes possible.

FIG. 11 shows how the cell group 104 placed in the recess 2 is sucked by the instrument 6. At this time, since the contaminants 105 have settled on the bottom surface of the recess 2, they are placed at a certain distance from the cell group 104 with respect to the suction port of the instrument 6. Therefore, when the cell group 104 is aspirated, the frequency of suctioning the contaminants 105 at the same time can be reduced.

As explained above, according to this embodiment, the following effects can be obtained.

(1) Contaminants 105 and cell group 104 are separated by structure 1 placed in cell storage container 100. Thereby, when the cell group 104 is aspirated from the cell storage container 1, it is possible to reduce the frequency of suctioning the contaminants 105 at the same time.

(2) By adjusting the size of the structures 1 and the spacing between the structures 1, it is possible to change the size of the contaminants 105 to be separated. For example, when a cell group 104 with a diameter of about 300 μm is housed in the cell storage container 100 together with a liquid material 101 such as a culture medium, the columnar structures 1 are supported in the recess 2 so that the spacing between the structures 1 is 50 μm. When placed on the surface 20, impurities 105 such as dead cells smaller than 50 μm fall between the structures 1, and the cell group 104 is placed on the structure 1. As a result, the physical positions of the cell group 104 and the contaminants 105 are separated from each other in the recess 2, so that when the cell group 104 is aspirated with the instrument 6, it is possible to reduce the degree of contamination by the contaminants 105.

[Production method]
A method of manufacturing a cell storage container 100 in which the structure 1 is disposed on the support surface 20 and a method of manufacturing a cell storage container 100 in which the structure 1 is not adhered to the support surface 20 will be described. FIG. 12 is a cross-sectional view showing a method for manufacturing a cell storage container.

A method for manufacturing the cell container 100 in which the structure 1 is adhered to the support surface 20 will be described using FIG. 12(a). First, an intaglio molding plate 106 corresponding to the shape in which the structure 1 is placed on the support surface 20 of the recess 2 is created. Next, resin or metal is poured into the molding intaglio 106. The resin material used for manufacturing is a material that hardens when left for a predetermined time or more, a photocurable resin that hardens by ultraviolet irradiation, or a thermosetting resin that hardens by heating. By peeling off the molding intaglio 106 after the resin has hardened, it is possible to obtain the cell storage container 100 in which the structure 1 is adhered to the support surface 20.

A method for manufacturing the cell storage container 100 in which the structure 1 is not bonded to the support surface 20 will be described using FIG. 12(b). First, a molding intaglio 106 corresponding to the recess 2 of the cell storage container 100 is created. Next, resin or metal is poured into the molding intaglio 106. The resin material used for manufacturing is a material that hardens when left for a predetermined time or more, a photocurable resin that hardens by ultraviolet irradiation, or a thermosetting resin that hardens by heating. After the resin is cured, the molding intaglio 106 is peeled off. Further, the structure 1 is created by cutting a thread-like (linear) structure having an arbitrary diameter into an arbitrary length. The above-described structure 1 is placed in the recess 2 of the resin member obtained by peeling off the molding intaglio 106. Thereby, a cell storage container 100 in which the structure 1 is not adhered to the support surface 20 of the recess 2 can be obtained.

This embodiment can be modified and implemented as follows. 14 to 23 are cross-sectional views showing modified examples of the cell storage container. The following modifications can be implemented in combination with each other within a technically consistent range.

As shown in FIG. 13, the structure 1 may have a cylindrical shape. As a result, the structure has fewer corners, and physical damage to cells is reduced, making it possible to suppress a decrease in cell activity.

As shown in FIG. 14, the structure 1 may have a prismatic shape. This makes it possible to arrange the gaps between the structures 1 in any size on the bottom surface of the recess 2, and adjust the ease of trapping the foreign objects 105 depending on the size of the foreign objects 105. It becomes possible to do so.

As shown in FIG. 15, the structure 1 may have a spherical (granular) shape. As a result, the gaps between the structures 1 become smaller than when the columnar structures 1 are arranged, so that smaller contaminants 105 can be trapped in the gaps. At this time, the length l indicates the diameter of the structure 1.

As shown in FIG. 16, the structure 1 may have a disk shape. As a result, the structures 1 overlap each other, so that the gaps between the structures 1 become very small, making it difficult for the contaminants 105 trapped in the gaps to be released into the liquid. This makes it possible to suppress the degree to which contaminants 105 are mixed in when cells are aspirated.

As shown in FIG. 17, the structure 1 may have a bowl shape (cup shape). As a result, the surface of the structure 1 becomes a curved surface, so that the foreign matter 105 easily slides on the surface of the structure 1, and the foreign matter 105 tends to move toward the bottom surface of the recess 2. By directing the contaminants 105 toward the bottom surface of the recess 2, it is possible to suppress the degree to which the contaminants 105 are mixed in when cells are aspirated.

As shown in FIG. 18, the structure 1 may have a donut shape (annular shape). As a result, the surface of the structure 1 becomes a curved surface, so that the foreign matter 105 easily slides on the surface of the structure 1, and the foreign matter 105 tends to move toward the bottom surface of the recess 2. By directing the contaminants 105 toward the bottom surface of the recess 2, it is possible to suppress the degree to which the contaminants 105 are mixed in when cells are aspirated.

As shown in FIG. 19, the structure 1 may have a band-like shape. This makes it easier for the water flow in the recess 2 to change, making it possible to suppress the frequency at which contaminants 105 are sucked in when sucking cells.

As shown in FIG. 20, the structure 1 may have grooves on its surface. This increases the surface area of the structure 1 and creates fine voids, making it possible to capture finer contaminants 105.

As shown in FIG. 21, the structure 1 may have a bent portion. This increases the voids formed between the structures 1, making it possible to trap large contaminants 105 such as dead cells.

As shown in FIG. 22, the structure 1 may have a branched portion. This increases the voids formed between the structures 1, making it possible to trap large contaminants 105 such as dead cells.

As shown in FIG. 23, the length of the structure 1 may be larger than the diameter of the suction port of the device for aspirating cells. This makes it possible to suppress the frequency at which the structure 1 is sucked into the device when sucking the cell group 104, and when the cell group 104 is sucked and used for another purpose, it is possible to suppress the frequency at which the structure 1 is sucked into the device. This makes it possible to prevent contamination. In addition, in the circular structure 1 as shown in FIGS. 15 to 18, the length l is the diameter of the structure 1, and in the structure 1 having branches as shown in FIG. 20 side and the end of the structure 1 extending from the branch part furthest from the support surface 20. Further, when a plurality of structures 1 are arranged in the recess 2, the sizes and shapes of all the structures 1 may be the same or different.

The cell storage container according to the present invention described in the above embodiment and the method for using the same are as follows.

[1] A cell storage container having a recess for storing cells,
A cell storage container, wherein a plurality of structures made of resin or metal, which have a specific gravity greater than 1, are water-insoluble, and are biocompatible, are arranged on the bottom surface of the recess.

[2] The structure is one or more materials selected from the group consisting of polylactide (PLA), polycaprolactone (PCL), polyglycolide (PGA), and polyesters based on copolymers thereof, or a plurality of materials. The cell storage container according to item 1, which is composed of a combination of materials.

[3] The cell storage container according to item 1 or 2, wherein the structure has an arithmetic mean roughness (Ra) of 0.03 μm or less.

[4]
The cell storage container according to any one of items 1 to 3, wherein the structure has been subjected to a surface treatment so that it does not adhere to cells.

[5] The cell storage container according to any one of items 1 to 4, wherein the structure has a cylindrical shape.

[6] According to any one of items 1 to 5, the maximum length of a straight line connecting the ends of the structure is equal to or larger than the diameter of the suction port of a device for aspirating cells. cell storage container.

[7] The cell storage container according to any one of items 1 to 6, wherein the structure has a bent portion.

[8] The cell storage container according to any one of items 1 to 7, wherein the structure has a branched portion.

[9] A method of using the cell storage container according to items 1 to 8, comprising:
discharging cells together with a liquid into the recess, and arranging the cells on the structure disposed on the bottom of the recess;
After allowing enough time for the water surface of the liquid to become flat and impurities to precipitate,
A method of using a cell storage container to aspirate the cells.

100...Cell storage container 1...Structure 2...Recess 5...Tray 6...Instrument 20...Support surface 50...Culture recess 101...Liquid 104...Cell group 105...Contaminants 106...Intaglio for molding

Claims (10)

A cell storage container having a recess for storing cells,
A cell storage container, wherein a plurality of structures made of resin or metal, which have a specific gravity greater than 1, are water-insoluble, and are biocompatible, are arranged on the bottom surface of the recess. The structure is made of one or more materials selected from the group consisting of polylactide (PLA), polycaprolactone (PCL), polyglycolide (PGA), and polyesters based on copolymers thereof. The cell storage container according to claim 1, which is comprised of a combination. The cell storage container according to claim 1, wherein the arithmetic mean roughness (Ra) of the structure is 0.03 μm or less. The cell storage container according to claim 1, wherein the structure has been subjected to a surface treatment that makes it non-adhesive to cells. The cell storage container according to claim 1, wherein the structure has a cylindrical shape. The cell storage container according to claim 1, wherein the maximum length of a straight line connecting the ends of the structure is equal to or longer than the diameter of a suction port of a device for suctioning cells. The cell storage container according to claim 1, wherein the structure has a bent portion. The cell storage container according to claim 1, wherein the structure has a branched portion. A method of using the cell storage container according to any one of claims 1 to 8, comprising:
discharging cells together with a liquid into the recess, and arranging the cells on the structure disposed on the bottom of the recess;
After allowing enough time for the water surface of the liquid to become flat and impurities to precipitate,
A method of using a cell storage container to aspirate the cells. The structure is made of one or more materials selected from the group consisting of polylactide (PLA), polycaprolactone (PCL), polyglycolide (PGA), and polyesters based on copolymers thereof, or a plurality of materials. 2. The cell storage container according to claim 1, wherein the cell container has an arithmetic mean roughness (Ra) of 0.03 μm or less and has been subjected to a surface treatment so as to be non-adhesive to cells.