Cell culture device

Abstract

The utility model relates to the technical field of biological experiment equipment, in particular to a cell culture device. The culture plate or the culture cup is provided with a gas-liquid interaction channel, the gas-liquid interaction channel is communicated with the culture hole, and the culture hole is communicated with the liquid storage tank through the gas-liquid interaction channel, so that the culture medium in the liquid storage tank can enter the culture hole through the gas-liquid interaction channel. Due to the fact that the culture plate or the culture cup is detachably arranged on the liquid storage plate, when a culture medium needs to be replaced, the culture plate or the culture cup can be separated from the liquid storage plate, then the culture medium in the liquid storage groove of the liquid storage plate is poured out, rapid replacement of the culture medium is achieved, and the problem that an existing cell culture plate is long in liquid replacement time is solved. Due to the fact that the culture medium can automatically flow out of the gas-liquid interaction channel, the culture medium can be replaced more thoroughly, the residual quantity of the culture medium in different culture holes is consistent, the culture medium does not need to be sucked by a suction pipe, organoid damage is not prone to being caused, and the differentiation efficiency can be improved.

Description

Technical Field

[0001] The present application relates to the technical field of biological experimental equipment, and in particular to a cell culture device. Background Art

[0002] Organoids are tissue analogs with a certain spatial structure formed by culturing pluripotent stem cells or adult stem cells in vitro. They are one of the most groundbreaking cutting-edge technologies in the biomedical field in recent years. Organoids can simulate real organs in terms of cell type, tissue structure, function and gene expression, and can be stably expanded and cultured in vitro, providing an ideal model for simulating organ development, biomedical research, drug development and clinical precision medicine.

[0003] In the differentiation and culture process of organoids, ultra-low adsorption cell culture plates are needed. According to the different bottom shapes, they can be divided into flat bottom and round bottom (U-shaped bottom and V-shaped bottom). According to the specifications of the cell culture plates, they can be divided into 6-well, 12-well, 24-well, 48-well, 96-well, 384-well, 1536-well and other culture plates. The existing 3D organoid culture process usually digests human induced pluripotent stem cells into single cells, inoculates them into low-adsorption cell culture plates, and then adds culture medium containing different differentiation factors according to the type of differentiated organoids.

[0004] At present, in actual operation, it is necessary to manually use a gun tip to aspirate the culture medium from the culture wells of the cell culture plate one by one. Usually, it takes 0.5-1h to change the medium in a 96-well plate, which has the problem of long liquid change time. In the process of aspirating the culture medium, a lot of residual culture medium is left, and the volume of residual culture medium in different wells of the plate varies greatly, which easily leads to different degrees of dilution of the added differentiation factor concentration, resulting in low differentiation efficiency. Utility Model Content

[0005] The present application provides a cell culture device for improving the technical problems of long liquid replacement time and low differentiation efficiency in current cell culture plates.

[0006] According to the first aspect, an embodiment provides a cell culture device, comprising:

[0007] A liquid storage plate, the liquid storage plate having a liquid storage tank for storing a culture medium;

[0008] A culture plate, the culture plate having culture holes for culturing cells, the number of the culture holes being two or more, and the culture plate having a gas-liquid interaction channel for allowing gas and liquid to pass through but preventing cells from passing through;

[0009] The culture plate is detachably arranged on the liquid storage plate so that the culture plate can be removed from the liquid storage plate; at least a portion of the culture plate is located in the liquid storage tank, and the culture hole is connected to the liquid storage tank through the gas-liquid interaction channel, so that the culture medium in the liquid storage tank can enter the culture hole through the gas-liquid interaction channel.

[0010] Furthermore, in one embodiment, the culture plate includes a plate body and a gas-liquid interaction membrane, the plate body has a plate body hole, the gas-liquid interaction membrane is fixed at the bottom of the plate body hole and surrounds the culture hole with the plate body, and the membrane hole of the gas-liquid interaction membrane forms the gas-liquid interaction channel.

[0011] Furthermore, in one embodiment, the culture plate is integrally formed, and the gas-liquid interaction channel is a gas-liquid interaction hole located at the bottom of the culture hole.

[0012] Furthermore, in one embodiment, the culture plate includes a positioning plate and a culture cup separate from the positioning plate, the positioning plate has a positioning hole, the culture cup is positioned and inserted into the positioning hole, the inner hole of the culture cup forms the culture hole, and the positioning plate is configured on the liquid storage plate.

[0013] Furthermore, in one embodiment, the culture cup has a retaining edge at the mouth of the cup, and the culture cup is hung on the positioning plate through the retaining edge, or the culture cup is fixedly connected to the positioning plate.

[0014] Furthermore, in one embodiment, the culture plate has a supporting edge supported at the notch of the liquid storage tank, and the culture plate is used to be placed on the liquid storage plate.

[0015] In one embodiment, a cell culture device comprises:

[0016] A liquid storage plate, the liquid storage plate having a liquid storage tank for storing a culture medium;

[0017] A culture cup, wherein the culture cup has culture holes for culturing cells, the number of the culture cups is two or more, and the culture cup has a gas-liquid interaction channel for allowing gas and liquid to pass through but preventing cells from passing through;

[0018] The culture cup is detachably arranged on the liquid storage plate so that the culture cup can be removed from the liquid storage plate; at least a portion of the culture cup is located in the liquid storage tank, and the culture hole is connected to the liquid storage tank through the gas-liquid interaction channel, so that the culture medium in the liquid storage tank can enter the culture hole through the gas-liquid interaction channel.

[0019] Furthermore, in one embodiment, the culture cup has a retaining edge at the mouth of the cup, and the culture cup is hung on the liquid storage plate through the retaining edge.

[0020] Furthermore, in one embodiment, the bottom of the culture well has the gas-liquid interaction channel.

[0021] Furthermore, in one embodiment, the gas-liquid interaction channel is a gas-liquid interaction hole, and the pore size of the gas-liquid interaction hole is 0.5-2 μm.

[0022] According to the cell culture device of the above embodiment, the culture plate or culture cup has a gas-liquid interaction channel, the gas-liquid interaction channel is connected to the culture well, and the culture well is connected to the liquid reservoir through the gas-liquid interaction channel, so that the culture medium in the liquid reservoir can enter the culture well through the gas-liquid interaction channel. Since the culture plate or culture cup can be detachably configured on the liquid reservoir plate, when the culture medium needs to be replaced, the culture plate or culture cup can be separated from the liquid reservoir plate, and then the culture medium in the liquid reservoir of the liquid reservoir plate can be poured out, so as to achieve rapid replacement of the culture medium, thereby improving the problem of long liquid replacement time of the current cell culture plate. Since the culture medium can automatically flow out from the gas-liquid interaction channel, the culture medium can be replaced more thoroughly, the residual amount of culture medium in different culture wells is consistent, and there is no need to use a pipette to aspirate the culture medium, which is not easy to cause damage to the organoids and can improve the differentiation efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 An exploded view of a cell culture device in one embodiment;

[0024] Figure 2 is a cross-sectional view of a cell culture device in one embodiment;

[0025] Figure 3 A partial cross-sectional view of a culture plate of another structure in an embodiment;

[0026] Figure 4 is a cross-sectional view of a plate body and a gas-liquid interaction membrane in one embodiment;

[0027] Figure 5 FIG. 4 is a cross-sectional view of a cell culture device in another embodiment.

[0028] List of feature names corresponding to the figure marks in the figure: 1. Liquid storage plate; 11. Liquid storage tank; 2. Culture plate; 21. Culture hole; 22. Gas-liquid interaction channel; 23. Support edge; 24. Positioning plate; 25. Culture cup; 251. Guard edge; 26. Plate body; 27. Gas-liquid interaction membrane; 3. Cover plate.

[0029] Explanation on the reference numerals in brackets in the drawings: In the reference numerals in brackets in the drawings, the features referred to by the reference numerals are both the features represented by the numbers in the brackets and the features represented by the numbers outside the brackets. DETAILED DESCRIPTION

[0030] The present application is further described in detail below by specific embodiments in conjunction with the accompanying drawings. Wherein similar elements in different embodiments adopt associated similar element numbers. In the following embodiments, many detailed descriptions are intended to enable the present application to be better understood. However, those skilled in the art can easily recognize that some of the features can be omitted in different situations, or can be replaced by other elements, materials, and methods. In some cases, some operations related to the present application are not shown or described in the specification, in order to avoid the core part of the present application being overwhelmed by too much description, and for those skilled in the art, it is not necessary to describe these related operations in detail, and they can fully understand the related operations based on the description in the specification and the general technical knowledge in the art.

[0031] In addition, the features, operations or characteristics described in the specification can be combined in any appropriate manner to form various implementations. At the same time, the steps or actions in the method description can also be interchanged or adjusted in a manner that is obvious to those skilled in the art. Therefore, the various sequences in the specification and the drawings are only for the purpose of clearly describing a certain embodiment and are not meant to be a required sequence, unless otherwise specified that a certain sequence must be followed.

[0032] The serial numbers of the components in this document, such as "first", "second", etc., are only used to distinguish the objects described and do not have any order or technical meaning. The "connection" and "coupling" mentioned in this application, unless otherwise specified, include direct and indirect connections (couplings).

[0033] Please refer to Figures 1 to 5 In one embodiment, the cell culture device includes a liquid storage plate 1 and a culture plate 2, wherein the liquid storage plate 1 has a liquid storage tank 11 for storing culture medium, and the culture plate 2 has culture wells 21 for culturing cells. The number of the culture wells 21 is more than two, and the culture plate 2 has a gas-liquid interaction channel 22 for allowing gas and liquid to pass through and preventing cells from passing through.

[0034] The culture plate 2 is detachably arranged on the liquid storage plate 1, so that the culture plate 2 can be removed from the liquid storage plate 1. At least part of the culture plate 2 is in the liquid storage tank 11, and the culture wells 21 and the liquid storage tank 11 can have gas-liquid interaction through the gas-liquid interaction channel 22, so that the culture medium in the liquid storage tank 11 can enter the culture wells 21 through the gas-liquid interaction channel 22, and of course, the culture medium in the culture wells 21 can also enter the liquid storage tank 11.

[0035] The culture plate 2 has a gas-liquid interaction channel 22, which is connected to the culture hole 21, and the culture hole 21 is connected to the liquid reservoir 11 through the gas-liquid interaction channel 22, so that the culture medium in the liquid reservoir 11 can enter the culture hole 21 through the gas-liquid interaction channel 22. By making the culture plate 2 detachable and configured on the liquid reservoir plate 1, when the culture medium needs to be replaced, the culture plate 2 can be separated from the liquid reservoir plate 1, and then the culture medium in the liquid reservoir 11 of the liquid reservoir plate 1 can be poured out, so as to realize the rapid replacement of the culture medium. Since the culture medium can automatically flow out from the gas-liquid interaction channel 22, the replacement is more thorough, and the residual amount of culture medium in different culture holes 21 is consistent, and there is no need to suck the culture medium with a pipette.

[0036] The culture plate 2 and the liquid storage plate 1 can be detachable in any feasible manner. For example, in one embodiment, please refer to Figure 1 The culture plate 2 has a supporting edge 23 supported at the notch of the liquid storage tank 11, and the culture plate 2 is used to be placed on the liquid storage plate 1. For another example, in one embodiment, please refer to Figure 5 , the culture plate 2 includes a positioning plate 24 and a culture cup 25, the positioning plate 24 has a positioning hole for positioning the culture cup 25, the inner hole of the culture cup 25 forms a culture hole 21, and the edge of the positioning plate 24 is supported at the notch of the liquid storage tank 11. For another example, in one embodiment, the bottom of the culture plate 2 contacts the bottom of the liquid storage plate 1, and the culture plate 2 is directly placed on the bottom surface of the liquid storage tank 11 of the liquid storage plate 1. For another example, in one embodiment, the culture plate 2 and the liquid storage plate 1 are detachably fixed to the liquid storage plate 1 by means of magnetic attraction, fasteners (such as screws), etc.

[0037] In one embodiment, please refer to Figures 1 to 3 , the culture plate 2 is integrally formed, and the gas-liquid interaction channel 22 is a gas-liquid interaction hole at the bottom of the culture hole 21. Specifically, in one embodiment, the gas-liquid interaction channel 22 at the bottom of the culture hole 21 is relatively small in size. In order to improve the processing accuracy, the culture plate 2 is formed by 3D printing. In some other embodiments, the culture plate 2 can also be formed by injection molding.

[0038] In one embodiment, since the 3D printing molding materials are expensive, in order to reduce the cost, in one embodiment, please refer to Figure 5The culture plate 2 includes a positioning plate 24 and a culture cup 25. The positioning plate 24 has a positioning hole for positioning the culture cup 25. The culture cup 25 is inserted into the positioning hole to ensure the stability of the culture cup 25. The inner hole of the culture cup 25 forms a culture hole 21, and the positioning plate 24 is arranged on the liquid storage plate 1. The culture cup 25 is separated from the positioning plate 24, which makes it convenient to process the culture cup 25 separately, and the positioning plate 24 can be produced by ordinary technology. In one embodiment, the positioning plate 24 is fixedly connected to the culture cup 25. The positioning plate 24 and the culture cup 25 can be fixedly connected by any feasible fixed connection method such as welding, bonding, clamping or fasteners.

[0039] In one embodiment, please refer to Figure 5 The culture cup 25 has a retaining edge 251 at its mouth, and the culture cup 25 is hung on the positioning plate 24 via the retaining edge 251 .

[0040] In addition to directly processing the gas-liquid interaction channel 22, in one embodiment, please refer to Figure 4 The culture plate 2 includes a plate body 26 and a gas-liquid interaction membrane 27. The plate body 26 has a plate body hole. The gas-liquid interaction membrane 27 is fixed to the bottom of the plate body hole and surrounds the plate body 26 to form a culture hole 21. The membrane hole of the gas-liquid interaction membrane 27 forms a gas-liquid interaction channel 22. Specifically, in one embodiment, please refer to Figure 4 The gas-liquid interactive membrane 27 is bonded or welded to the plate 26 .

[0041] In addition to using a culture plate 2 configured on the liquid storage plate 1, in one embodiment, the cell culture device includes a liquid storage plate 1 and a culture cup 25, the liquid storage plate 1 has a liquid storage tank 11 for storing culture medium, the culture cup 25 has a culture well 21 for culturing cells, the number of the culture cups 25 is more than two, and the culture cup 25 has a gas-liquid interaction channel 22 for allowing gas and liquid to pass through and preventing cells from passing through. The culture cup 25 is detachably configured on the liquid storage plate 1 so that the culture cup 25 can be removed from the liquid storage plate 1. At least part of the culture cup 25 is in the liquid storage tank 11, and the culture well 21 is connected to the liquid storage tank 11 through the gas-liquid interaction channel 22, so that the culture medium in the liquid storage tank 11 can enter the culture well 21 through the gas-liquid interaction channel 22.

[0042] In one embodiment, the mouth of the culture cup 25 has a retaining edge 251 , and the culture cup 25 is hung on the liquid storage plate 1 via the retaining edge 251 .

[0043] In one embodiment, in order to make the culture medium replacement more thorough, a gas-liquid interaction channel 22 is provided at the bottom of the culture well 21. Of course, in addition to the bottom of the culture well 21, a gas-liquid interaction channel 22 may also be provided in the middle of the wall of the culture well 21.

[0044] The number of gas-liquid interaction channels 22 can be increased or decreased as needed. In one embodiment, the number of gas-liquid interaction channels 22 can be any number of one, two, or more than three. In one embodiment, the gas-liquid interaction channels 22 can be gas-liquid interaction holes or through grooves that penetrate the gas-liquid interaction holes.

[0045] In one embodiment, please refer to Figure 3 The gas-liquid interaction channel 22 is a gas-liquid interaction hole, and the pore size of the gas-liquid interaction hole is 0.5-2 μm. Specifically, in one embodiment, the pore size of the gas-liquid interaction hole is 1.0 μm. In some other embodiments, the pore size of the gas-liquid interaction hole can be increased or decreased as needed, for example, the pore size of the gas-liquid interaction hole can be

[0046] In one embodiment, the liquid storage plate 1 and the culture plate 2 are both rectangular plates. In some other embodiments, in addition to rectangular plates, the liquid storage plate 1 and the culture plate 2 can also be oval plates, circular plates, or any other feasible shapes.

[0047] In one embodiment, please refer to Figure 2 In order to allow the culture medium to smoothly enter the culture wells 21, a gap is left between the outermost culture wells 21 and the wall of the liquid reservoir 11. A gap is also left between the bottom surface of the culture plate 2 and the bottom surface of the liquid reservoir 11. Specifically, in one embodiment, please refer to Figure 2 , the distance d1 between the bottom surface of the culture plate 2 and the bottom surface of the liquid storage tank 11 is 2.5 mm. Figure 2 , the distance d2 between the outer wall surface of the outermost culture hole 21 and the side wall of the liquid storage tank 11 is 5mm-6mm. In one embodiment, the inner diameter of the culture hole 21 can be 5mm-7mm, the outer diameter of the culture hole 21 is 6mm-10mm, the wall thickness of the culture hole 21 is 0.5mm-3mm, and the height h1 of the culture hole 21 is 8mm-16mm.

[0048] In one embodiment, the culture wells 21 are evenly distributed on the culture plate 2. The culture plate 2 uses an ultra-low adsorption culture plate to prevent the 3D organoids from adhering to the well walls of the culture wells 21. The number of culture wells 21 on the culture plate 2 can be any number greater than two, such as 6 wells, 12 wells, 24 wells, 48 ​​wells, 96 wells, 384 wells, 1536 wells, etc.

[0049] In one embodiment, please refer to Figure 1 and Figure 2 The culture plate 2 and the liquid storage plate 1 are both made of biocompatible materials, such as optically transparent pure polystyrene.

[0050] In one embodiment, the cell culture device includes a cover plate 3 , which is used to cover the liquid storage plate 1 and can cover the culture wells 21 on the culture plate 2 .

[0051] In one embodiment, the cell culture device is used for 3D organoid culture, please refer to Figures 1 to 5 , the steps for using the cell culture device are as follows:

[0052] In the biosafety cabinet, the same volume of cell suspension is added to the culture well 21 according to the experimental needs. Due to the influence of gravity, after several hours, the cells autonomously assemble into a 3D microsphere structure and settle to the bottom of the culture well 21, while the culture medium leaks from the gas-liquid interaction channel 22 into the liquid storage plate 1.

[0053] After the cell microspheres are formed, take out the culture plate 2 and place it on the cover plate 3, suck out the culture medium in the reservoir 11, and then add the corresponding volume of culture medium containing differentiation factors into the reservoir 11, place the culture plate 2 into the reservoir 11, and gently shake the reservoir plate 1 so that the culture medium is completely distributed in the reservoir 11. The liquid level of the added culture medium is higher than the height of the cell microspheres in the culture well 21.

[0054] Cover the cover plate 3 and place it in an incubator. During the subsequent culture process, the culture medium is replaced according to the type of differentiated organoids.

[0055] The above specific examples are used to illustrate the present application, which is only used to help understand the present application and is not intended to limit the present application. For technicians in the technical field to which the present application belongs, they can also make some simple deductions, deformations or substitutions based on the ideas of the present application.

Claims

1. A cell culture device, characterized in that: include: A liquid storage plate, the liquid storage plate having a liquid storage tank for storing a culture medium; A culture plate, the culture plate having culture holes for culturing cells, the number of the culture holes being two or more, and the culture plate having a gas-liquid interaction channel for allowing gas and liquid to pass through but preventing cells from passing through; The culture plate is detachably arranged on the liquid storage plate so that the culture plate can be removed from the liquid storage plate; at least a portion of the culture plate is located in the liquid storage tank, and the culture hole and the liquid storage tank can perform gas-liquid interaction through the gas-liquid interaction channel, so that the culture medium in the liquid storage tank can enter the culture hole through the gas-liquid interaction channel.

2. The cell culture device according to claim 1, characterized in that The culture plate comprises a plate body and a gas-liquid interaction membrane, wherein the plate body has a plate body hole, the gas-liquid interaction membrane is fixed at the bottom of the plate body hole and surrounds the culture hole with the plate body, and the membrane hole of the gas-liquid interaction membrane forms the gas-liquid interaction channel.

3. The cell culture device according to claim 1, characterized in that: The culture plate is integrally formed, and the gas-liquid interaction channel is a gas-liquid interaction hole located at the bottom of the culture hole.

4. The cell culture device according to claim 1, characterized in that: The culture plate includes a positioning plate and a culture cup separated from the positioning plate. The positioning plate has a positioning hole. The culture cup is positioned and inserted into the positioning hole. The inner hole of the culture cup forms the culture hole. The positioning plate is arranged on the liquid storage plate.

5. The cell culture device according to claim 4, characterized in that: The culture cup has a retaining edge at the cup mouth, and the culture cup is hung on the positioning plate through the retaining edge, or the culture cup is fixedly connected to the positioning plate.

6. The cell culture device according to claim 1, characterized in that: The culture plate has a supporting edge supported at the notch of the liquid storage tank, and the culture plate is used to be placed on the liquid storage plate.

7. A cell culture device, characterized in that: include: A liquid storage plate, the liquid storage plate having a liquid storage tank for storing a culture medium; A culture cup, wherein the culture cup has culture holes for culturing cells, the number of the culture cups is two or more, and the culture cup has a gas-liquid interaction channel for allowing gas and liquid to pass through but preventing cells from passing through; The culture cup is detachably arranged on the liquid storage plate so that the culture cup can be removed from the liquid storage plate; at least a portion of the culture cup is located in the liquid storage tank, and the culture hole is connected to the liquid storage tank through the gas-liquid interaction channel, so that the culture medium in the liquid storage tank can enter the culture hole through the gas-liquid interaction channel.

8. The cell culture device according to claim 7, characterized in that: The culture cup has a retaining edge at its cup mouth, and the culture cup is hung on the liquid storage plate through the retaining edge.

9. The cell culture device according to any one of claims 1 to 8, characterized in that: The bottom of the culture well is provided with the gas-liquid interaction channel.

10. The cell culture device according to any one of claims 1 to 8, characterized in that: The gas-liquid interaction channel is a gas-liquid interaction hole, and the pore size of the gas-liquid interaction hole is 0.5-2 μm.

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