Cell culture device capable of conveniently changing liquid

Abstract

The utility model relates to a cell culture device capable of conveniently changing liquid, which comprises a culture plate, a liquid collecting plate and a liquid changing plate which are respectively provided with a plurality of culture bins, a plurality of liquid collecting holes and a plurality of liquid changing holes, and has a liquid collecting state, a liquid discharging state and a liquid changing state. In the liquid collecting state, the culture plates are stacked on the liquid collecting plates, so that each culture bin is contained in the corresponding liquid collecting hole, and a protruding part arranged at the bottom of each liquid collecting hole is embedded into and blocks a through opening formed in the bottom wall of the corresponding culture bin. The culture plate is lifted from the liquid collecting plate so that the protruding parts of the liquid collecting holes are drawn out from the through openings; in the liquid changing state, the culture plates after liquid discharging are stacked on the liquid changing plate, so that each culture bin is contained in the corresponding liquid changing hole, and the new culture liquid in the liquid changing hole enters the culture bin through the through opening of the culture bin. According to the cell culture device, rapid liquid change can be realized without a pipettor, and adverse effects on adherent cells caused by liquid change of the pipettor are avoided.

Description

Technical Field

[0001] The utility model relates to the technical field of cell culture, in particular to a cell culture device capable of conveniently changing liquid. Background Art

[0002] Cell culture plates are experimental tools used to culture cells in vitro. They are used for cell research and provide an environment suitable for cell growth. Depending on the purpose of the experiment, culture plates of corresponding specifications can be selected to facilitate multi-condition and multi-group cell experimental research. The advantages of cell culture plates include providing controllable conditions, facilitating cell observation, and adapting to the culture of various cell types. Usually, cell experiments involve multiple operations such as changing fluids and adding drugs.

[0003] When the cell culture plate of the prior art is undergoing a liquid change operation, the cover of the cell culture plate is removed, the old culture solution in each culture well is sucked out one by one by a pipette, and then new culture solution is added to each well. On the one hand, when the waste liquid is sucked out with a pipette, the pipette tip is prone to accidentally touch the bottom surface of the cell culture, causing the adherent cells to fall off, and it is also prone to incomplete absorption, resulting in residual waste liquid; when a new culture solution is added with a pipette, the flow rate of the sampled liquid is too fast, causing the adherent cells to be blown up. On the other hand, when changing the liquid, in order to prevent the pipette tip from accidentally touching the bottom surface of the cell culture and to avoid the adherent cells being blown up by too fast a liquid flow rate, the operator usually slows down the operation process by slowly adding liquid along the side wall of the cell culture well. At the same time, in order to prevent cross contamination, the pipette tip is frequently replaced. Such a liquid change operation process is too cumbersome, the operation time is too long, and it is time-consuming and laborious. After the cell culture wells absorb the waste liquid, they may not be able to add new culture fluid in time, causing the cells to dehydrate for a long time and die; the long operation time will also cause a time difference in the injection time of each cell culture well, resulting in inconsistent culture time of each well, affecting the uniformity of the experimental data, and unable to guarantee the repeatability of the experimental results. Moreover, the cell culture cover plate needs to be removed throughout the liquid replacement operation, and the excessive operation time increases the risk of cell contamination. In addition, when conducting cell experiments in vitro, it is sometimes necessary to set up cell groups with different culture times. When the number of experimental wells is not large, if multiple well plates are operated, it will cause a waste of resources and increase the burden of medical waste disposal. Therefore, laboratories usually set up multiple culture time groups on a cell culture well plate, but this will cause the well plate to be repeatedly taken out of the incubator for liquid replacement operations, and the culture wells that do not need to be replaced will also be repeatedly taken out for opening operations, increasing the risk of contamination; due to the different culture time points, it is also inconvenient to observe cells and collect samples. Utility Model Content

[0004] The purpose of the present invention is to solve at least one of the above problems and / or other defects in the prior art.

[0005] To achieve the above-mentioned purpose, according to one aspect of the utility model, a cell culture device capable of conveniently changing liquid is provided, which comprises: a culture plate, on which a plurality of culture chambers for culturing cells are arranged, and the bottom wall of each culture chamber is provided with at least one through opening; a liquid collecting plate, on which a plurality of liquid collecting holes corresponding to the plurality of culture chambers are arranged, and the bottom of each liquid collecting hole is provided with at least one protrusion extending toward the inside of the liquid collecting hole and matching with the through opening of the corresponding culture chamber; and a liquid changing plate, on which a plurality of liquid changing holes corresponding to the plurality of culture chambers for holding new culture liquid are arranged. The cell culture device is configured to work in a liquid collecting state, a liquid releasing state and a liquid changing state according to the positional relationship of the culture plate relative to the liquid collecting plate or the liquid changing plate. In the liquid collecting state, the culture plate is stacked on the liquid collecting plate so that each culture bin is received in the corresponding liquid collecting hole and the protrusion of the liquid collecting hole is embedded in and blocks the through opening of the culture bin, and in the liquid discharging state, the culture plate is lifted from the liquid collecting plate so that the protrusion of the liquid collecting hole is drawn out from the through opening to allow the waste culture fluid in the culture bin to be discharged into the liquid collecting hole through the through opening; in the liquid changing state, the culture plate after liquid discharge is stacked on the liquid changing plate so that each culture bin is received in the corresponding liquid changing hole and the new culture fluid in the liquid changing hole enters the culture bin through the through opening of the culture bin.

[0006] According to one embodiment of the utility model, the culture bin is cylindrical with a circular flat bottom wall, the at least one through opening includes two arc-shaped openings extending circumferentially along the bottom wall of the culture bin and radially opposite to each other, and the at least one protrusion includes two arc-shaped protrusions respectively matched with the two arc-shaped openings.

[0007] According to one embodiment of the utility model, a sealing ring is fixed around the two arc-shaped protrusions on the inner side of the bottom of the liquid receiving hole, and the inner diameter of the sealing ring matches the outer diameter of the culture bin so that when the liquid receiving hole receives the corresponding culture bin, the sealing ring abuts against the outer wall of the culture bin from the radial outside.

[0008] According to one embodiment of the utility model, the culture plate is provided with a plurality of placement holes for respectively placing the plurality of culture chambers, and the top of the culture chamber is provided with a peripheral edge portion extending radially outward, and when the culture chamber is placed through the corresponding placement holes, the peripheral edge portion abuts against the top surface of the culture plate.

[0009] According to one embodiment of the present invention, the culture chamber can be detachably or permanently placed in the corresponding placement hole.

[0010] According to one embodiment of the utility model, the cell culture device also includes a cover plate, the cover plate has a downwardly bent periphery, the top of the culture plate has a culture plate boss for arranging the multiple culture chambers, and a culture plate platform edge is arranged around the culture plate boss for accommodating the periphery of the cover plate when the cover plate covers the culture plate.

[0011] According to one embodiment of the utility model, when the cover plate covers the culture plate, there is a gap between the periphery of the cover plate and the outer peripheral side of the culture plate boss, which connects the culture chamber on the culture plate boss with the outside atmosphere.

[0012] According to one embodiment of the utility model, the cover plate and the culture plate boss are respectively provided with cover plate cut corners and culture plate cut corners that match each other at their respective corresponding corners.

[0013] According to one embodiment of the present invention, the bottom wall of the culture chamber has an inner surface that has been subjected to a surface treatment capable of improving cell adhesion.

[0014] According to one embodiment of the utility model, the culture plate has a peripheral edge bent downward, the top of the liquid receiving plate and the top of the liquid exchange plate respectively have a liquid receiving plate boss and a liquid exchange plate boss for setting the multiple liquid receiving holes and the multiple liquid exchange holes, and a liquid receiving plate platform edge and a liquid exchange plate platform edge are respectively arranged around the liquid receiving plate boss and the liquid exchange plate boss for accommodating the peripheral edge of the culture plate when the culture plate is respectively stacked on the liquid receiving plate and the liquid exchange plate.

[0015] The convenient cell culture device for changing liquid of the utility model can be used for the whole process of changing liquid. The operator only needs to hold the culture plate together with the culture chamber and pick it up from the liquid collecting plate, wait for a while, and then directly put the culture chamber on the culture plate into the liquid changing hole filled with new culture liquid after the waste culture liquid in the culture chamber is discharged into the liquid collecting hole through the through opening in the bottom wall of the culture chamber. There is no need to use a pipette to perform the liquid changing operation, which avoids the pipette tip accidentally touching the adherent cells in the culture chamber and causing the cells to fall off, and also avoids the excessively fast liquid flow rate when the pipette is adding the sample and blowing up the adherent cells. The whole liquid changing operation is simple and fast, which overcomes the disadvantage of low efficiency caused by the need to use a pipette to slowly suck out the waste liquid and then slowly add liquid along the side wall when changing the liquid of a conventional cell culture plate, and greatly improves the efficiency of the cell culture plate. The time for changing the liquid is shortened, the work efficiency is improved, and the long-term dehydration and death of cells caused by the failure to add new culture medium in time due to the long liquid change time is avoided; there is no need to open the cover of the culture plate when changing the liquid, which avoids the risk of cell contamination due to opening the cover; rapid liquid change avoids the problem of inconsistent sample addition time of each group, which is convenient for ensuring the uniformity of experimental data and the repeatability of experimental results; the whole set of equipment has a simple structure and low production cost, and can provide a variety of specifications and styles. It can be flexibly selected and used according to experimental needs, and can also be mass-produced, which is convenient for promotion and use; it can realize large-scale batch liquid change processing of cell culture plates, improve work efficiency, promote the standardization of the liquid change process, and improve the reliability of the experimental process, providing conditions for the implementation of cell culture automation technology in the future. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] The features and advantages of the present invention will be clearly understood through the detailed description provided below with reference to the accompanying drawings. It should be understood that the following drawings are merely schematic and not necessarily drawn to scale, and thus cannot be regarded as limiting the present invention, wherein:

[0017] Figure 1 1 is an overall schematic diagram of the components of a cell culture device according to an embodiment of the present invention, wherein the bottommost diagram represents both a liquid collection plate and a liquid exchange plate.

[0018] Figure 2 yes Figure 1 A schematic diagram of a culture plate in a cell culture apparatus is shown.

[0019] Figure 3 yes Figure 2 An enlarged schematic diagram of the culture chamber on the culture plate is shown.

[0020] Figure 4 yes Figure 1 Schematic diagram of the collection plate in the cell culture apparatus is shown.

[0021] Figure 5 yes Figure 4 An enlarged schematic diagram of the liquid collecting holes on the liquid collecting plate is shown.

[0022] Figure 6 yes Figure 1 Schematic diagram of the fluid exchange plate in the cell culture device shown.

[0023] Figure 7 yes Figure 6 An enlarged schematic diagram of the fluid exchange holes on the fluid exchange plate is shown.

[0024] Description of reference numerals:

[0025] 1. Culture plate; 11. Culture chamber; 111. Through opening; 112. Peripheral edge; 12. Culture plate boss; 13. Culture plate corner cut; 14. Culture plate platform edge; 2. Liquid collecting plate; 21. Liquid collecting hole; 211. Protrusion; 22. Sealing ring; 23. Liquid collecting plate boss; 24. Liquid collecting plate platform edge; 3. Liquid exchange plate; 31. Liquid exchange hole; 32. Liquid exchange plate boss; 34. Liquid exchange plate platform edge; 4. Cover plate; 41. Cover plate corner cut. DETAILED DESCRIPTION

[0026] Embodiments of the present invention are described below with reference to the accompanying drawings. In the following description, many specific details are set forth so that those skilled in the art can more fully understand and implement the present invention. However, it is obvious to those skilled in the art that the implementation of the present invention may not have some of these specific details. In addition, it should be understood that the present invention is not limited to the specific embodiments described. On the contrary, any combination of the features and elements described below may be considered to implement the present invention, regardless of whether they relate to different embodiments. Therefore, the following aspects, features, embodiments and advantages are for illustrative purposes only and should not be regarded as elements or limitations of the claims unless clearly stated in the claims.

[0027] Figure 1 FIG. 2 shows a schematic diagram of a cell culture device that can facilitate fluid replacement according to an embodiment of the present invention. Figure 1 As shown, the cell culture device according to this embodiment may include a cover plate 4, a culture plate 1, a liquid collecting plate 2 and a liquid exchange plate 3, wherein the overall appearance of the liquid collecting plate 2 and the liquid exchange plate 3 may be completely the same, so Figure 1 The cell culture experiment involves the operation of changing the medium. Figure 1As shown, before the liquid is replaced, the cell culture device may be stacked with the cover plate 4, the culture plate 1 and the liquid receiving plate 2 from top to bottom, and after the liquid is replaced, the cover plate 4, the culture plate 1 and the liquid changing plate 3 may be stacked from top to bottom. The culture plate 1 is provided with a plurality of culture chambers 11, and the liquid receiving plate 2 and the liquid changing plate 3 are respectively provided with a corresponding number of liquid receiving holes 21 and liquid changing holes 31, so that before the liquid is replaced, when the culture plate 1 is stacked on the liquid receiving plate 2, each culture chamber 11 can be received in each liquid receiving hole 21 in a one-to-one correspondence, and after the liquid is replaced, when the culture plate 1 is stacked on the liquid changing plate 3, each culture chamber 11 can be received in each liquid changing hole 31 in a one-to-one correspondence. It should be pointed out that the directional terms such as "upper", "lower", "top", "bottom" mentioned in this article refer to the placement posture of the cell culture device and its components under normal use (for example Figure 1 The posture shown is described by

[0028] Figure 1 Each of the cover plate 4, the culture plate 1, the liquid collecting plate 2 and the liquid exchanging plate 3 can be formed of a plate having a thickness of, for example, 1 to 2 mm and is generally rectangular in shape as a whole. Those skilled in the art will appreciate that, depending on the number and / or arrangement of the culture chambers 11 provided on the culture plate 1, the cover plate 4, the culture plate 1, the liquid collecting plate 2 and the liquid exchanging plate 3 can also be designed into other shapes.

[0029] Reference Figure 1 According to an exemplary configuration, the cover plate 4 may have a length of 127 to 128 mm and a width of 85 to 86 mm, and its periphery may be bent downward, thereby forming an overall height of 6 to 9 mm, so that it can be reliably covered on the culture plate 1 to provide good protection for it. The cover plate 4 is provided with a cut corner (referred to herein as a "cover plate cut corner") 41 for distinguishing the direction at one of its corners, so that the operator can easily find the correct orientation of the cover plate 4 when covering the culture plate 1, and quickly and accurately assemble the cover plate 4 on the culture plate 1.

[0030] Figure 2 The culture plate 1 according to one embodiment is shown in more detail. Figure 2As shown, the culture plate 1 may have, for example, a length of 127 to 128 mm, a width of 85 to 86 mm, and a height of 9 to 12 mm, and may be provided with a flat boss (referred to herein as "culture plate boss") 12 with a height of, for example, 2 to 3 mm on its top as its main body for arranging a plurality of culture chambers 11. Accordingly, a circle of edge portions (referred to herein as "culture plate edge portions") 14 is provided around the culture plate boss 12, which is used to receive the periphery of the cover plate 4 when the cover plate 4 covers the culture plate 1, and may, for example, have a width of 1 to 2 mm and a height of 2 to 3 mm (the same height as the culture plate boss 12). Providing the culture plate edge portion 14 to receive the periphery of the cover plate 4 is conducive to reducing the total height of the cover plate 4 when it is stacked on the culture plate 1, making the entire device compact and regular. Advantageously, when the cover 4 covers the culture plate 1, the bottom end of the peripheral edge of the cover 4 is supported on the top surface of the culture plate platform edge 14, and there is a gap between the inner peripheral side surface of the peripheral edge of the cover 4 (facing the culture plate boss 12) and the corresponding outer peripheral side surface of the culture plate boss 12. This gap can connect each culture chamber 11 on the culture plate boss with the outside atmosphere, so that the culture chamber 11 is in a semi-closed state, ensuring that the temperature, humidity and gas concentration inside the culture chamber 11 are consistent with those of the outside world (such as a cell culture box). In this way, the risk of bacterial contamination can be prevented by the cover 4, and gas exchange between the culture chamber 11 and the outside world can be achieved.

[0031] The culture plate boss 12 is also provided with a cut corner (referred to herein as "culture plate cut corner") 13 at its corner corresponding to the position of the cover plate cut corner 41 for easy identification of direction. The culture plate cut corner 13 and the cover plate cut corner 41 are adapted to each other. Only when the cover plate 4 and the culture plate 1 are in the correct orientation can the cover plate 4 be accurately assembled on the culture plate 1 through the cooperation of the cover plate cut corner 41 and the culture plate cut corner 13 to form a good coverage of the culture plate 1.

[0032] The culture plate 1 also has a peripheral edge bent downward so as to be stacked and assembled on the liquid collecting plate 2 or the liquid changing plate 3 (to be further described below). The peripheral edge of the culture plate 1 can be touched by the operator to facilitate taking the culture plate 1. For this purpose, the peripheral edge of the culture plate 1 can preferably be formed with anti-slip grooves on all sides (see Figure 2 The length of the culture plate 1 may be 60 to 70 mm, and the width may be 6 to 7 mm, so as to prevent the culture plate 1 from accidentally slipping during the process of taking the culture plate 1 and causing the experiment to fail.

[0033] The culture bins 11 may be arranged on the culture plate 1 in a split or integrated manner. The split type means that each culture bin 11 is used separately from the culture plate 1, and each culture bin 11 can be placed separately on the culture plate 1, which is convenient for disassembly and adjustment. A corresponding number of culture bins 11 can be placed according to the experimental design needs, which is convenient for adding samples, observing and collecting samples of different groups and different culture times. The integrated type means that each culture bin 11 is connected and combined with the culture plate 1 or is originally formed as a whole during manufacturing, which can realize the overall placement of each culture bin, without the need to place the culture bins one by one, and the operation is more convenient, saving time and effort. In order to place each culture bin 1, a plurality of placement holes (not shown) may be correspondingly arranged on the culture plate 1. The culture bin 11 can be detachably or permanently placed in the corresponding placement hole, thereby realizing the above-mentioned split or integrated arrangement respectively.

[0034] exist Figure 2 and 3 In the illustrated embodiment, the culture bin 11 is cylindrical with a circular flat bottom wall, and the corresponding placement hole is also circular. The diameter of the placement hole is, for example, 20 to 21 mm, and the diameter of each culture bin 11 is slightly smaller than the diameter of the placement hole, for example, 18 to 19 mm. The height of the culture bin 11 is, for example, 21 to 25 mm, and the volume can be 6 to 7 mL. The culture bin 11 can be provided with a peripheral edge 112 extending radially outward at its top, and the peripheral edge has, for example, a width of 3 to 4 mm and a thickness of 1 to 2 mm. Thus, when the culture bin 11 is placed through the corresponding placement hole, the peripheral edge 112 can abut against the top surface of the culture plate 1 to prevent the culture bin from falling. In addition, the peripheral edge 112 can be easily clamped, which is beneficial to the placement and removal operation of the culture bin 11 on the culture plate 1.

[0035] Continue to refer Figure 3 , the bottom wall of the culture bin 11 may be provided with at least one through opening 111, so that the waste culture fluid to be replaced can be discharged from the culture bin 11 through the through opening 111 without tilting or inverting the culture bin 11 when changing the fluid. In the illustrated embodiment, the at least one through opening 111 includes two arc-shaped openings extending circumferentially along the bottom wall of the culture bin 11 and radially opposite to each other, which is conducive to providing a complete and large area of ​​cell attachment on the bottom wall of the culture bin as much as possible and emptying the original waste culture fluid as much as possible when changing the fluid. In one embodiment, the width of the arc-shaped opening is 1 to 2 mm, the central angle is about 120°, and the arc length is 12 to 18 mm. Those skilled in the art will understand that the through opening 111 is not limited to the above configuration, but may also have other shapes, sizes, quantities or distribution methods, as long as the waste culture fluid can be discharged from the culture bin 11.

[0036] The inner wall surface of the culture chamber 11, especially the inner surface of the bottom wall, may be subjected to, for example, tissue culture (TC) treatment or other similar surface treatments to improve the adhesion of various cell types and facilitate cell adhesion, growth and extension.

[0037] Figure 4 An embodiment of the liquid collecting plate 2 is shown in more detail. The liquid collecting plate 2 may have a length of 127 to 128 mm, a width of 85 to 86 mm, and a height of 20 to 21 mm, and may be provided with a plurality of liquid collecting holes 21 corresponding to the plurality of culture chambers 11 on the culture plate 1, for receiving the original waste culture fluid in the culture chambers 11 when replacing the fluid. Figure 4 As shown, a boss (referred to herein as "liquid collecting plate boss") 23 with a height of, for example, 7 to 8 mm is provided on the top of the liquid collecting plate 2 as its main body for setting a plurality of liquid collecting holes 21. Accordingly, a circle of edge portions (referred to herein as "liquid collecting plate edge portions") 24 are provided around the liquid collecting plate boss 23, which are used to receive the periphery of the culture plate 1 when the culture plate 1 is stacked on the liquid collecting plate 2, and may, for example, have a width of 1 to 2 mm and a height of 7 to 8 mm (the same height as the liquid collecting plate boss 23). Providing the liquid collecting plate edge portion 24 to receive the periphery of the culture plate 1 is conducive to reducing the total height of the culture plate 1 when it is stacked on the liquid collecting plate 2, making the entire device compact and regular.

[0038] When the culture plate 1 is stacked on the liquid receiving plate 2, the center position of each liquid receiving hole 21 is consistent with the center position of the placement hole of the corresponding culture chamber 11 on the culture plate 1, and each culture chamber 11 can be received in the corresponding liquid receiving hole 21. For example, the inner diameter of the liquid receiving hole 21 is 22 to 23 mm, the hole depth and inner height are 17 to 18 mm, and the hole volume is 6 to 7 mL.

[0039] like Figure 5 As shown, the bottom of each liquid receiving hole 21 is also configured as a flat-bottomed circle that matches the culture chamber 11, and may be provided with at least one protrusion 211 extending toward the inside of the liquid receiving hole, and the shape of each protrusion 211 matches the shape of a corresponding through opening 111 of the corresponding culture chamber 11. For example, Figure 5 The bottom of the liquid receiving hole 21 shown is provided with two arc-shaped protrusions extending in the circumferential direction of the liquid receiving hole and arranged opposite to each other in the radial direction. For example, each arc-shaped protrusion has a height of 3 to 4 mm, a width of 1 to 2 mm, a central angle of about 120°, and an arc length of 12 to 18 mm, so that the arc-shaped protrusion can be similar in shape and size to the liquid receiving hole 21. Figure 3Due to this adaptability, when the culture bin 11 is received in the corresponding liquid receiving hole 21, each protrusion 211 of the liquid receiving hole can be embedded in the corresponding through opening on the culture bin and block the through opening to prevent the culture liquid in the culture bin from flowing out through the through opening. When the through opening 111 adopts other shapes, sizes, quantities or distribution methods other than the above configuration, the protrusions 211 are correspondingly set to a matching shape, size, quantity or distribution method.

[0040] Optionally, from Figure 5 It can also be seen that a sealing ring 22 is provided on the inner side of the bottom of the liquid receiving hole 21. The sealing ring 22 is arranged on the radial outer side of the two arc-shaped protrusions 211, and its inner diameter is set to match the outer diameter of the culture chamber 11. In this way, when the culture plate 1 is stacked on the liquid receiving plate 2 so that the liquid receiving hole 21 receives the corresponding culture chamber 11, the sealing ring 22 can abut against the outer wall of the culture chamber 11 from the radial outer side (the abutting position is located at the bottom wall of the culture chamber 11 in the vertical direction), thereby producing a sealing effect on the culture chamber 11, thereby more reliably preventing the culture liquid in the culture chamber 11 from flowing out. For example, the inner diameter of the sealing ring 22 is 18 to 19 mm, the outer diameter is 1 to 2 mm wider than the inner diameter, and the height or thickness is 0.5 to 1 mm. The material of the sealing ring 22 is, for example, rubber, which has a certain elasticity, and can be fixed on the inner side of the bottom of the liquid receiving hole 21, for example, by gluing.

[0041] Figure 6 An embodiment of the liquid exchange plate 3 is shown in more detail. The liquid exchange plate 3 may have, for example, a length of 127 to 128 mm, a width of 85 to 86 mm, and a height of 20 to 21 mm, and may be provided with a plurality of liquid exchange holes 31 corresponding to the plurality of culture chambers 11 on the culture plate 1, for receiving new culture fluid to be replaced into the culture chambers 11 during liquid exchange. Figure 6 As shown, a boss (referred to herein as "the boss of the liquid exchange plate") 32 with a height of, for example, 7 to 8 mm is provided on the top of the liquid exchange plate 3 as its main body for setting a plurality of liquid exchange holes 31. Accordingly, a circle of edge portions (referred to herein as "the edge portions of the liquid exchange plate") 34 is provided around the liquid exchange plate boss 32, which is used to receive the periphery of the culture plate 1 when the culture plate 1 is stacked on the liquid exchange plate 3, and may, for example, have a width of 1 to 2 mm and a height of 7 to 8 mm (the same height as the boss of the liquid exchange plate 32). Providing the edge portions 34 of the liquid exchange plate to receive the periphery of the culture plate 1 is also conducive to reducing the total height of the culture plate 1 when it is stacked on the liquid exchange plate 3, making the entire device compact and regular.

[0042] When the culture plate 1 is stacked on the liquid exchange plate 3, the center position of each liquid exchange hole 31 is consistent with the center position of the placement hole of the corresponding culture chamber 11 on the culture plate 1, and each culture chamber 11 can be received in the corresponding liquid exchange hole 31. For example, Figure 7 The inner diameter of the fluid exchange hole 31 is 22 to 23 mm, the hole depth and inner height are 17 to 18 mm, and the hole volume is 6 to 7 mL. Figure 4 , 5 and Figure 6 , 7 From the comparison, it can be seen that, except that the protrusion 211 and the sealing ring 22 are provided in the liquid receiving hole 21 of the liquid receiving plate 2, the liquid exchange plate 3 and its liquid exchange hole 31 can be completely the same as the liquid receiving plate 2 and its liquid receiving hole 21.

[0043] In the illustrated embodiment, the number of the culture chamber 11 and the corresponding liquid collection holes 21 and liquid exchange holes 31 on the culture plate 1 is twelve, but the present invention is not limited thereto. The number of the culture chamber and the liquid collection holes and liquid exchange holes can be appropriately selected according to the experimental purpose, including but not limited to six, twelve, twenty-four and ninety-six, etc., so as to facilitate multi-condition and multi-group cell experimental research. In addition, the size and arrangement of the culture chamber 11 and the corresponding liquid collection holes and liquid exchange holes are not limited to the above-described embodiments, but can adopt any appropriate design according to the experimental requirements.

[0044] The above-mentioned components of the cell culture device of the embodiment of the utility model can be made of plastic, for example, polystyrene with extremely high transparency can be used, and its light transmittance can reach more than 90%, which is convenient for cell observation. In addition, the entire cell culture device can be sterilized by gamma rays or other sterilization treatments to make it sterile, pyrogen-free (i.e., endotoxin-free), DNase (i.e., deoxyribonuclease) and RNase (i.e., ribonuclease), avoiding bacterial contamination during the cell culture process.

[0045] The following briefly describes an exemplary working process of a cell culture device capable of conveniently changing fluids according to an embodiment of the present invention.

[0046] First, place the liquid collecting plate 2 on a stable operating table, and stack the culture plate 1 that has not been loaded with the culture chamber 11 on the liquid collecting plate 2, so that the multiple placement holes on the culture plate 1 are aligned one by one with the liquid collecting holes 21 on the liquid collecting plate 2. Then, according to the experimental design, the required number of culture chambers 11 are placed in the corresponding placement holes on the culture plate 1 and then received in the corresponding liquid collecting holes 21 on the liquid collecting plate 2. When placing the culture chambers, align the through opening 111 of each culture chamber 11 with the protrusion 211 in the corresponding liquid collecting hole 21 and embed the protrusion 211 into the corresponding through opening 111 to block the through opening, and at the same time place the bottom of the culture chamber 11 in the sealing ring 22 in the liquid collecting hole 21; when the peripheral edge 112 of the culture chamber 11 abuts against the top surface of the culture plate 1, the placement of the culture chamber is completed. Next, add the prepared cell suspension (volume of 1 mL, for example) into each culture chamber 11 by a pipette. Finally, the cover plate 4 is placed on the culture plate 1 by aligning the cover plate corner 41 with the culture plate corner 13, and the cover plate 4, culture plate 1 and liquid collecting plate 2 stacked on each other are placed in an incubator for cell culture. In this state, the protrusion 211 of the liquid collecting hole 21 is embedded in the through opening 111 of the culture bin adapted thereto, and the sealing ring 22 is provided, which can reliably prevent the cell suspension (or the initial culture fluid) in the culture bin from flowing out through the through opening 111, thereby ensuring that the cells can be well plated and inoculated in the culture bin. In this article, the working state or position in which the culture plate 1 is stacked on the liquid collecting plate 2 as described above so that each culture bin 11 is received in the corresponding liquid collecting hole 21 and the protrusion 211 of the liquid collecting hole is embedded in and blocks the through opening of the culture bin is referred to as the "liquid collecting state" of the cell culture device.

[0047] When it is necessary to change the liquid, first place the liquid change plate 3 on the operating table, and use a pipette to add the prepared new culture liquid (the volume is also, for example, 1 mL) to the corresponding liquid change hole 31 on the liquid change plate 3. Then, take out the culture plate 1 that needs to be changed from the incubator together with the liquid receiving plate 2 and the cover plate 4, and place them on the operating table. Next, directly pick up the culture plate 1 (with the culture chamber 11) and the cover plate 4 from the liquid receiving plate 2 and lift them to a certain height, so that the protrusion 211 of each liquid receiving hole 21 is pulled out from its corresponding through opening 111, and stay for a while so that the waste culture liquid in each culture chamber 11 is discharged into the liquid receiving hole 21 through the through opening 111 that is no longer blocked by the protrusion. This is referred to as "discharging" in this article, and this working state or position of the cell culture device is referred to as "discharging state" in this article. After all the waste culture fluid in each culture chamber 11 is discharged, the culture plate 1 with the empty culture chamber is stacked together with the cover plate 4 on the fluid exchange plate 3 placed on the operating table, so that each culture chamber 11 is received in the corresponding fluid exchange hole 31. At this time, the new culture fluid previously added to the fluid exchange hole 31 will naturally enter the empty culture chamber through the through opening 111 of the corresponding culture chamber, thereby realizing the rapid fluid exchange of each culture chamber. The working state or position in which the culture plate 1 after the fluid is discharged is stacked on the fluid exchange plate 3 so that each culture chamber 11 is received in the corresponding fluid exchange hole 31 and the new culture fluid in the fluid exchange hole enters the culture chamber through the through opening 111 of the culture chamber is referred to as the "fluid exchange state" of the cell culture device in this article.

[0048] As described above, the cell culture device for convenient fluid exchange of the present invention can use the protrusion in the fluid collection hole on the fluid collection plate to block the through opening in the bottom wall of the corresponding culture chamber on the culture plate in the fluid collection state to prevent the initial culture fluid in the culture chamber from flowing out, so as to achieve the initial plating and inoculation of cells in the culture chamber; when fluid exchange is required, the cell culture device can be switched to the fluid discharge state first, that is, the operator holds the culture plate with the culture chamber together with the cover plate and picks it up from the fluid collection plate and waits for a while, so that the waste culture fluid in the culture chamber is completely discharged into the fluid collection hole through the through opening in the bottom wall of the culture chamber; finally, the cell culture device enters the fluid exchange state, that is, the empty culture chamber on the culture plate after fluid discharge is directly placed into the fluid exchange hole filled with new culture fluid on the fluid exchange plate, so that the new culture fluid enters the culture chamber through the through opening again, thereby completing the fluid exchange. It can be seen that the cell culture device of the utility model has at least the following technical effects or advantages: there is no need to use a pipette when changing the liquid, which avoids the pipette tip accidentally touching the adherent cells in the culture chamber and causing the cells to fall off, and also avoids the liquid flow rate being too fast when the pipette is adding samples and blowing up the adherent cells; the entire liquid change operation is simple and fast, which overcomes the disadvantage of low efficiency caused by the need to use a pipette to slowly suck out the waste liquid and then slowly add liquid along the side wall when changing the liquid in a conventional cell culture plate, greatly shortens the liquid change time, improves work efficiency, and avoids the long time of cell growth caused by the inability to add new culture fluid in time due to the long liquid change time. Death from dehydration; There is no need to open the cover of the culture plate when changing the liquid, avoiding the risk of cell contamination due to opening the cover; Rapid liquid change avoids the problem of inconsistent sample addition time for each group, which is convenient for ensuring the uniformity of experimental data and the repeatability of experimental results; The whole set of equipment has a simple structure and low production cost, and can provide a variety of specifications and styles. It can be flexibly selected and used according to experimental needs, and can also be mass-produced, which is convenient for promotion and use; It can realize large-scale batch liquid change of cell culture plates, improve work efficiency, promote the standardization of the liquid change process, and improve the reliability of the experimental process, providing conditions for the implementation of cell culture automation technology in the future.

[0049] For those skilled in the art, various modifications and variations may be made to the embodiments disclosed above without departing from the scope or spirit of the utility model. Other embodiments of the utility model will be apparent to those skilled in the art from the practice of the utility model disclosed in this specification. This specification and the examples disclosed therein should be considered to be illustrative only, and the true scope of the utility model is specified by the appended claims and their equivalents.

Claims

1. A cell culture device capable of conveniently changing liquid, characterized in that include: A culture plate (1), wherein a plurality of culture chambers (11) for culturing cells are provided on the culture plate, and the bottom wall of each culture chamber is provided with at least one through opening (111); a liquid receiving plate (2), the liquid receiving plate being provided with a plurality of liquid receiving holes (21) respectively corresponding to the plurality of culture chambers (11), the bottom of each liquid receiving hole being provided with at least one protrusion (211) extending toward the inside of the liquid receiving hole and matching with the through opening (111) of the corresponding culture chamber; and a liquid exchange plate (3), wherein the liquid exchange plate is provided with a plurality of liquid exchange holes (31) respectively corresponding to the plurality of culture chambers (11) and used for containing new culture liquid, The cell culture device is configured to operate in a liquid collecting state, a liquid releasing state and a liquid changing state according to the positional relationship of the culture plate (1) relative to the liquid collecting plate or the liquid changing plate. In the liquid collecting state, the culture plate (1) is stacked on the liquid collecting plate (2) so that each culture chamber (11) is received in the corresponding liquid collecting hole (21) and the protrusion (211) of the liquid collecting hole is embedded in and blocks the through opening (111) of the culture chamber, and in the liquid discharging state, the culture plate (1) is lifted from the liquid collecting plate (2) so that the protrusion of the liquid collecting hole is withdrawn from the through opening to allow the waste culture fluid in the culture chamber to be discharged into the liquid collecting hole through the through opening; in the liquid changing state, the culture plate (1) after liquid discharging is stacked on the liquid changing plate (3) so that each culture chamber (11) is received in the corresponding liquid changing hole (31) and the new culture fluid in the liquid changing hole enters the culture chamber through the through opening (111) of the culture chamber.

2. The cell culture device according to claim 1, characterized in that: The culture bin (11) is cylindrical with a circular flat bottom wall, the at least one through opening (111) comprises two arc-shaped openings extending circumferentially along the bottom wall of the culture bin and radially opposite to each other, and the at least one protrusion (211) comprises two arc-shaped protrusions respectively matched with the two arc-shaped openings.

3. The cell culture device according to claim 2, characterized in that: A sealing ring (22) is fixed around the two arc-shaped protrusions on the inner side of the bottom of the liquid receiving hole (21), and the inner diameter of the sealing ring matches the outer diameter of the culture bin so that when the liquid receiving hole receives the corresponding culture bin, the sealing ring abuts against the outer wall of the culture bin from the radial outside.

4. The cell culture device according to any one of claims 1 to 3, characterized in that: The culture plate (1) is provided with a plurality of placement holes for respectively placing the plurality of culture chambers (11), and the top of the culture chamber is provided with a peripheral edge portion (112) extending radially outward, and when the culture chamber is placed through the corresponding placement holes, the peripheral edge portion abuts against the top surface of the culture plate (1).

5. The cell culture device according to claim 4, characterized in that: The culture chamber (11) can be detachably or permanently placed in the corresponding placement hole.

6. The cell culture device according to any one of claims 1 to 3, characterized in that: The cell culture device also includes a cover plate (4) having a peripheral edge bent downward, the top of the culture plate (1) having a culture plate boss (12) for arranging the multiple culture chambers (11), and a culture plate platform edge (14) is arranged around the culture plate boss for accommodating the peripheral edge of the cover plate when the cover plate covers the culture plate.

7. The cell culture device according to claim 6, characterized in that: When the cover plate (4) covers the culture plate (1), a gap exists between the periphery of the cover plate and the outer peripheral side of the culture plate boss (12) so that the culture chamber (11) on the culture plate boss is connected to the outside atmosphere.

8. The cell culture device according to claim 6, characterized in that: The cover plate (4) and the culture plate boss (12) are respectively provided with a cover plate cut corner (41) and a culture plate cut corner (13) adapted to each other at their respective corresponding corners.

9. The cell culture device according to any one of claims 1 to 3, characterized in that: The bottom wall of the culture chamber (11) has an inner surface that has been subjected to a surface treatment capable of improving cell adhesion.

10. The cell culture device according to any one of claims 1 to 3, characterized in that: The culture plate (1) has a peripheral edge bent downward, and the top of the liquid receiving plate (2) and the top of the liquid exchange plate (3) respectively have a liquid receiving plate boss (23) and a liquid exchange plate boss (32) for setting the multiple liquid receiving holes and the multiple liquid exchange holes, and a liquid receiving plate platform edge (24) and a liquid exchange plate platform edge (34) are respectively arranged around the liquid receiving plate boss and the liquid exchange plate boss for accommodating the peripheral edge of the culture plate when the culture plate is respectively stacked on the liquid receiving plate and the liquid exchange plate.

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