Tumor organ culture dish

Abstract

The utility model discloses a tumor organ culture dish which comprises a shell and a culture unit, and the culture unit comprises a first culture chamber, a second culture chamber and a pipetting tank; the first culture chamber is a cavity formed in the shell; the second culture chamber is connected with the shell, the second culture chamber is communicated with the first culture chamber, and the pipetting tank is communicated with the first culture chamber. The tumor organ culture dish provided by the utility model is simple in structure and convenient to operate, only the old culture medium in the lower chamber needs to be sucked and discarded from the pipetting tank, and a new culture medium needs to be added from the pipetting tank again, so that the gas-liquid culture and liquid change operation of tumor organs is facilitated; transfer of the culture room is avoided during liquid change, and observation of the growth state of a fixed area in the tumor organ culture process is facilitated; an additional culture unit required for transferring the culture room during liquid change in the traditional operation is reduced, and the cost is reduced.

Description

Technical Field

[0001] The utility model relates to the technical field of in vitro organ culture, in particular to a tumor organoid culture dish. Background Art

[0002] Tumor organoids are organoids obtained by culturing tumor patient biopsy, puncture or surgical resection tissue in matrix gel for several weeks. While maintaining the heterogeneity of the source tumor and the heterogeneity between patients, tumor organoids maintain basically uniform morphology and scale between individual organoids, providing a fast and excellent technical platform for tumor pathogenesis research, drug screening, personalized precision medicine, regenerative medicine and other fields. In recent years, with the continuous development and innovation of in vitro organ culture technology, more and more three-dimensional in vitro tumor models containing more immune cells are being designed.

[0003] At present, tumor organoid culture models mainly include the following four types: 1. Directly adding exogenous immune cells to the traditional organoid culture system to simulate the interaction between tumor cells and specific types of immune cells; 2. Using transwell cell culture plates to culture tumor tissues by air-liquid interface (ALI), which can preserve the original immune cells in the tumor tissue, better simulate the tumor microenvironment, and retain more original structural distribution characteristics; 3. While adding exogenous immune cells, the 3D-microfusion technology can also retain tumor-infiltrating lymphocytes; 4. 3D bioprinting technology that uses tissue engineering technology to flexibly reconstruct the tumor immune microenvironment, etc.

[0004] The transwell cell culture plate includes an upper chamber (small chamber) and a lower chamber. In the ALI tumor organoid culture model, the organoids in the upper chamber are regularly observed and photographed. There are the following problems in the operation of replacing the culture medium in the lower chamber of the transwell: First, during the liquid replacement operation, it is necessary to clamp the upper chamber (small chamber) to another culture well, then aspirate and discard the culture medium in the lower chamber of the original culture well, and add new culture medium; or add fresh culture medium to the new culture well, then clamp the upper chamber (small chamber) to the new well containing fresh culture medium, and finally aspirate and discard the old culture medium. This liquid replacement step is cumbersome and easily introduces the risk of falling and contamination during the process of clamping and replacing the upper chamber (small chamber). Second, due to the liquid replacement operation, the position of the small chamber is changed, which is not convenient for observing the growth status of tumor organoids in a fixed position. Utility Model Content

[0005] The utility model aims to provide a tumor organoid culture dish, which does not need to transfer the culture chamber when replacing the culture medium, is easy to operate, and can effectively avoid contamination of the cultured organ during the process of replacing the culture medium.

[0006] To achieve the above-mentioned purpose, the technical solution adopted by the utility model is to provide a tumor organoid culture dish, including a shell and at least one culture unit, the culture unit including a first culture chamber, a second culture chamber and a pipette groove; the first culture chamber is a cavity formed on the shell; the second culture chamber is connected to the shell, the second culture chamber is located in the first culture chamber and is connected to the first culture chamber, and the pipette groove is arranged on the shell and is connected to the first culture chamber.

[0007] Furthermore, the second culture chamber is fixedly connected to the shell via a connecting bridge.

[0008] Furthermore, the main body of the second culture chamber is in a ring shape, and a polyester film is arranged at the bottom of the second culture chamber.

[0009] Furthermore, the polyester film is a polycarbonate film or a polyethylene terephthalate film, and the pore size of the polyester film is 0.4 μm.

[0010] Furthermore, the pipetting groove is an oblique groove opened on the shell, and the bottom of the pipetting groove is connected to the first culture chamber.

[0011] Furthermore, the oblique groove is a tapered groove that is wider at the top and narrower at the bottom.

[0012] Furthermore, a gap is formed between the bottom of the second culture chamber and the bottom of the first culture chamber, and the gap height is 3-5 mm.

[0013] Further, the top edge of the shell is kept flush with or slightly higher than the top edge of the second culture chamber.

[0014] Furthermore, the culture dish includes a plurality of culture units, and the plurality of culture units are arranged in a matrix in the shell.

[0015] Furthermore, the number of the culture units is m*n, which are arranged in a matrix of m rows and n columns, where m and n are both positive integers, m is 2-8, and n is 2-12.

[0016] Compared with the prior art, the utility model has the following beneficial effects: the tumor organoid culture dish provided by the utility model has a simple structure and is easy to operate by arranging a pipette groove connected to the first culture chamber (lower chamber) on the shell. When replacing the culture medium, it is only necessary to suck out the old culture medium in the first culture chamber (lower chamber) from the pipette groove and re-add new culture medium from the pipette groove, which is convenient for the gas-liquid culture liquid replacement operation of tumor organoids and can effectively avoid the contamination of the cultured organ in the process of replacing the culture chamber and the culture medium in the traditional operation; since there is no need to clamp and transfer the second culture chamber (upper chamber, small chamber) to the new culture hole when changing the liquid, the second culture chamber can be fixedly connected to the shell, and then the growth status and the like can be observed in a fixed area during the tumor organoid culture process; in addition, the culture dish provided by the utility model reduces the need for additional culture units to transfer the second culture chamber when changing the liquid in the traditional operation, thereby reducing costs. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is a schematic diagram of the structure of a tumor organoid culture dish in an embodiment of the utility model;

[0018] Figure 2 It is a top view schematic diagram of a tumor organoid culture dish in an embodiment of the utility model;

[0019] Figure 3 for Figure 2 Schematic diagram of the cross section along the CC;

[0020] FIG4 is a schematic diagram of the structure of a tumor organoid culture dish with 6, 12, and 24 culture units, respectively, in an embodiment of the present invention;

[0021] In the figure:

[0022] 1-shell; 2-culture unit; 21-first culture chamber; 22-second culture chamber; 3-connecting bridge; 4-polyester film; 5-pipette groove. DETAILED DESCRIPTION

[0023] The utility model is further described below in conjunction with the accompanying drawings and embodiments.

[0024] See also Figure 1 , Figure 2 and Figure 3This embodiment provides a tumor organoid culture dish, including a shell 1 and at least one culture unit 2, the culture unit 2 includes a first culture chamber 21, a second culture chamber 22 and a pipetting groove 5, the first culture chamber 21 is a cavity formed on the shell 1, the second culture chamber 22 is located in the first culture chamber 21 and is fixedly connected to the shell 1, the second culture chamber 22 is connected to the first culture chamber 21, so that the culture medium can flow between the second culture chamber 22 and the first culture chamber 21. The shell 1 is provided with a pipetting groove 5, which is connected to the first culture chamber 21, so that the culture medium can be sucked away from the first culture chamber 21 or put into the culture medium through the pipetting groove 5.

[0025] Preferably, the main body of the second culture chamber 22 is annular, and a permeable polyester film 4 is provided at the bottom of the second culture chamber 22, and the second culture chamber 22 and the first culture chamber 21 penetrate the culture medium through the polyester film 4. The polyester film 4 is preferably a polycarbonate film or a polyethylene terephthalate film, and the pore size of the polyester film 4 is preferably 0.4 μm. The second culture chamber 22 is fixedly connected to the shell 1 through a connecting bridge 3. Specifically, at least two connecting bridges 3 are provided between the second culture chamber 22 and the shell 1, and one end of each connecting bridge 3 is connected to the outer wall of the second culture chamber 22, and the other end of the connecting bridge 3 is connected to the inner wall of the shell 1. Preferably, a plurality of connecting bridges 3 are evenly spaced along the circumferential direction, and the number of connecting bridges 3 is preferably two. Furthermore, the connecting bridge 3, the second culture chamber 22, and the shell 1 are integrally formed, which can be integrally formed by a mold, or by 3D printing, etc.

[0026] Preferably, the first culture chamber 21 is a round hole, and there is a gap between the bottom of the second culture chamber 22 and the bottom of the first culture chamber 21, and the gap is not less than 3mm, preferably 3mm-5mm; the distance between the outer wall of the second culture chamber 22 and the inner wall of the shell 1 is preferably 2mm-5mm. The culture dish also includes a lid (not shown), and the top edge of the second culture chamber 22 should be flush with the top edge of the shell 1 or slightly lower than the top edge of the first culture chamber 21, so as to facilitate the closing of the culture dish lid.

[0027] In this embodiment, the second culture chamber 22 is fixedly connected to the inner wall of the shell 1 through the connecting bridge 3, so that when changing the liquid, it is only necessary to tilt the culture dish, and the pipette can suck out the old culture medium from the pipette groove 5 and then inject new culture medium, thereby avoiding the movement of the second culture chamber 22 and facilitating the observation of the growth status of a fixed area during the tumor organoid culture process.

[0028] The tumor organoid culture dish provided by the utility model can be provided with a plurality of culture units 2. Preferably, the plurality of culture units 2 are arranged in a matrix of m rows and n columns in the shell 1, where m and n are both positive integers, and the number of culture units 2 is m*n. For example, 6 (2*3), 12 (3*4) and 24 (4*6) culture units 2 are respectively taken as examples. Figure 4a , Figure 4b and Figure 4c The arrangement shown is that in other embodiments, those skilled in the art can design the arrangement as needed.

[0029] The use process of the tumor organoid culture dish provided by the utility model is as follows:

[0030] 1. Add pre-cooled matrix gel into the second culture chamber 22 for coating, and incubate in a 37° C. incubator for 30 minutes to wait for the matrix gel layer to solidify.

[0031] 2. Continue to add pre-cooled tissue gel containing tumor tissue and matrix gel into the second culture chamber 22, and incubate in a 37°C incubator for 30 minutes to wait for the tissue gel to solidify.

[0032] 3. Use a pipette to add tumor organoid culture medium into the pipetting trough 5 so that the liquid level of the culture medium in the first culture chamber 21 is slightly lower than or flush with the height of the matrix gel layer in the second culture chamber 22.

[0033] 4. Place the culture dish in a 37°C, 5% CO2 incubator.

[0034] 5. Change the medium every 2-3 days according to the growth status of the tumor organoids. Tilt the culture dish toward the pipette trough 5, use the pipette tip to suck out the old culture medium from the pipette trough 5, and then add fresh tumor organoid culture medium from the pipette trough 5.

[0035] In summary, the tumor organoid culture dish provided by the utility model has a simple structure and is easy to operate by arranging a pipette groove connected to the first culture chamber on the shell. When replacing the culture medium, it is only necessary to suck out the old culture medium in the lower chamber from the pipette groove and re-add new culture medium from the pipette groove, which is convenient for the gas-liquid culture fluid replacement operation of tumor organoids and can effectively avoid the contamination of the cultured organs during the replacement of culture chambers and culture media in traditional operations; since there is no need to clamp and transfer the second culture chamber to the new culture chamber when changing the fluid, the second culture chamber can be fixedly connected to the shell, and then the growth status and the like can be observed in a fixed area during the tumor organoid culture process; in addition, the culture dish provided by the utility model reduces the need for additional culture units to transfer the second culture chamber when changing the fluid in traditional operations, thereby reducing costs.

[0036] Although the present invention has been disclosed as above with preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art may make some modifications and improvements without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention shall be based on the definition of the claims.

Claims

1. A tumor organoid culture dish, characterized in that: It includes a shell and at least one culture unit, the culture unit includes a first culture chamber, a second culture chamber and a pipette groove; the first culture chamber is a cavity formed on the shell; the second culture chamber is connected to the shell, the second culture chamber is located in the first culture chamber and communicated with the first culture chamber, and the pipette groove is arranged on the shell and communicated with the first culture chamber.

2. The tumor organoid culture dish according to claim 1, characterized in that: The second culture chamber is fixedly connected to the shell via a connecting bridge.

3. The tumor organoid culture dish according to claim 1, characterized in that: The second culture chamber body is in a ring shape, and a polyester film is arranged at the bottom of the second culture chamber.

4. The tumor organoid culture dish according to claim 3, characterized in that: The polyester film is a polycarbonate film or a polyethylene terephthalate film, and the pore size of the polyester film is 0.4 μm.

5. The tumor organoid culture dish according to claim 1, characterized in that: The pipetting groove is an oblique groove opened on the shell, and the bottom of the pipetting groove is communicated with the first culture chamber.

6. The tumor organoid culture dish according to claim 5, characterized in that: The oblique groove is a tapered groove that is wider at the top and narrower at the bottom.

7. The tumor organoid culture dish according to claim 1, characterized in that: A gap is formed between the bottom of the second culture chamber and the bottom of the first culture chamber, and the height of the gap is 3-5 mm.

8. The tumor organoid culture dish according to claim 1, characterized in that: The top edge of the shell is flush with or slightly higher than the top edge of the second culture chamber.

9. The tumor organoid culture dish according to claim 1, characterized in that: The culture dish comprises a plurality of culture units, and the plurality of culture units are arranged in a matrix in the shell.

10. The tumor organoid culture dish according to claim 9, characterized in that: The number of the culture units is m*n, which are arranged in a matrix with m rows and n columns, and m and n are both positive integers, m is 2-8, and n is 2-12.

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