Microporous plate for 3D culture and function evaluation test of primary cells

Abstract

The utility model discloses a microwell plate for 3D (three-dimensional) culture and function evaluation test of primary cells, belongs to the technical field of microwell plates, and solves the problems that a plurality of tumor organs are different in size and are distributed on different planes due to the fact that the existing primary tumor organ cells are cultured in a plurality of culture dishes. The porous plate comprises a porous plate main body and a plurality of counter bores arranged on the porous plate main body in an array; a plurality of sinking grooves are formed in the bottom face of each counter bore, and each sinking groove is in an inverted regular rectangular pyramid shape. A plurality of tumor organs which are consistent in size and are positioned on the same height plane can be cultured by the plurality of counter bores in the microplate main body, so that subsequent imaging detection and statistics are facilitated. The arrangement of the inverted rectangular pyramid-shaped sinking groove enables primary tumor organoid cells to be uniformly gathered to the bottom of the sinking groove, formation of a more consistent organoid is facilitated, and the rectangular pyramid-shaped sinking groove can guide the organoid or cells to be concentrated at a specific position, so that improvement of imaging consistency is facilitated.

Description

Technical Field

[0001] The utility model relates to the technical field of microporous plates, in particular to a microporous plate used for primary cell 3D culture and function evaluation tests. Background Art

[0002] Primary cell 3D culture is an important technology in biomedical research, with broad application prospects and far-reaching significance. In tumor biology research, primary cell 3D culture can reveal tumor cell proliferation, metastasis, and drug sensitivity, providing an experimental basis for personalized cancer treatment. Clinical trials have shown that drug sensitivity testing using patient-derived tumor organoids can be closely correlated with the patient's actual clinical outcomes, providing important auxiliary information for clinical treatment. Therefore, using patient-derived tumor organoids to test the sensitivity of chemotherapy drugs, targeted drugs, new anti-tumor antibody drugs, etc. can provide strong support for doctors to develop personalized treatment plans.

[0003] However, currently used primary tumor organoid cultures are often cultured in culture dishes with matrigel. The culture dishes contain a specific organoid culture medium that is rich in growth factors to support the growth and expansion of primary tumor organoids. The cells of the tumor organoids will gradually form a three-dimensional structure within the matrigel in the culture medium. However, for drug sensitivity testing, tumor organoids cultured in multiple culture dishes will vary in size, making it difficult to form a standardized structure. This inconsistency will affect the accuracy of the test. At the same time, the distribution of tumor organoids on different planes also poses challenges to imaging detection and drug efficacy evaluation, thereby affecting the repeatability and comparability of experimental results. Utility Model Content

[0004] In response to the above-mentioned problems in the prior art, the present invention provides a microplate for 3D culture and functional evaluation of primary cells, which solves the problem that the existing primary tumor organoid cells are cultured in multiple culture dishes, resulting in multiple tumor organoids of different sizes and distributed on different planes.

[0005] In order to achieve the above purpose, the technical solution adopted by the utility model is as follows:

[0006] Provided is a microplate for primary cell 3D culture and functional evaluation tests, comprising a microplate body and a plurality of sink holes arrayed on the microplate body; a plurality of sink grooves are provided on the bottom surface of each sink hole, and each sink groove is in the shape of an inverted regular quadrangular pyramid.

[0007] The working principle of this scheme is to inoculate primary tumor organoid cells in multiple sinks and solidify them with matrix gel, and then add culture medium for cultivation, so that multiple sinks on the microplate body can culture tumor organoids of the same number and size, and multiple tumor organoids are located on the same height plane, which is convenient for subsequent imaging detection and statistics. The inverted tetrahedral sink setting allows primary tumor organoid cells to gather at the bottom of the sink, and matrix gel can be added to contact the cells, which is more conducive to promoting healthy cell growth and improving cell survival rate. The tetrahedral sink can also guide organoids or cells to concentrate in specific locations, preventing them from being scattered in different areas of the well and growing in fixed positions, thereby helping to improve the consistency of imaging and ensure that cells or tumor organoids can be accurately located in each sink in high-throughput screening for microscopic observation and imaging analysis, thereby improving the repeatability and comparability of drug sensitivity experiments.

[0008] Furthermore, a plurality of countersunk holes are arranged in a rectangular array. The rectangular array design ensures that the plurality of countersunk holes are evenly distributed on the microplate.

[0009] Furthermore, each well is square. This square well design provides better growth space and stability for primary tumor organoid cells during culture, helping to reduce drift during the culture process.

[0010] Furthermore, four sink grooves are provided in each countersunk hole, and the four sink grooves are arranged in a square array in the countersunk hole and cover the entire bottom of the countersunk hole, so that the four sink grooves can maximize the use of the space of the countersunk hole.

[0011] Furthermore, the side length of each well is 3mm to 4mm. This setting not only provides sufficient space for the growth of primary tumor organoid cells, but also ensures that the tumor organoids can fully contact the culture medium during drug sensitivity testing, promoting uniform drug distribution and improving detection sensitivity.

[0012] Furthermore, each side of each trough has an inclination angle of 45° to 75°. This inclination of the inner wall of the trough facilitates the sedimentation and positioning of primary tumor organoid cells in the trough, while also facilitating the flow of liquid in the culture medium, promoting the even distribution of nutrients, and reducing air bubble interference, further improving the growth quality of tumor organoids and the accuracy of drug response.

[0013] Furthermore, the microplate body includes a rectangular frame and a main plate fixed within the rectangular frame. The top of the rectangular frame is provided with end plates fixed to the main plate. The rectangular frame enhances the stability of the main plate and prevents deformation of the main plate due to external forces or manipulation during the experiment.

[0014] Furthermore, the four inner sides of the rectangular frame are fixedly connected to the four outer sides of the main board through multiple ribs. The hollowing out of the bottom of the rectangular frame reduces gravity, while the multiple ribs increase the connection strength between the rectangular frame and the main board.

[0015] Furthermore, outwardly protruding ridges are provided around the bottom of the rectangular frame, which facilitates the operation and movement of the microplate body during the experiment.

[0016] Furthermore, a positioning notch is provided at one corner of the rectangular frame. The positioning notch can help the experimenter quickly confirm the orientation of the microplate body, ensuring the consistency of the well position during each operation or data reading, and avoiding data confusion or experimental errors caused by incorrect orientation.

[0017] The utility model discloses a microplate for primary cell 3D culture and function evaluation test, which has the following beneficial effects:

[0018] The multiple wells on the microplate body of this utility model can cultivate multiple tumor organoids of uniform size and located at the same height plane, facilitating subsequent imaging detection and statistics. The inverted square pyramid-shaped sink allows primary tumor organoid cells to evenly aggregate at the bottom of the sink. This not only helps to form more consistent organoids compared to existing microplates without sinks, but the square pyramid sink can also guide the organoids or cells to concentrate in a specific location, preventing them from being scattered across different areas of the wells and growing in a fixed position, thereby helping to improve imaging consistency. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is the axonometric view of the microplate;

[0020] Figure 2 It is a top view of the microplate;

[0021] Figure 3 This is a bottom view of the sink;

[0022] Figure 4 This is a bottom view of the microplate;

[0023] Figure 5 This is the axonometric drawing of the mainboard;

[0024] Among them: 1. Main board; 11. Countersunk hole; 12. Countersunk groove; 2. Rectangular frame; 21. End plate; 22. Edge plate; 23. Rib plate. DETAILED DESCRIPTION

[0025] The specific implementation methods of the present invention are described below to facilitate understanding of the present invention by those skilled in the art. However, it should be clear that the present invention is not limited to the scope of the specific implementation methods. For those skilled in the art, as long as various changes are within the spirit and scope of the present invention as defined and determined by the appended claims, these changes are obvious, and all utility model creations using the concept of the present invention are protected.

[0026] refer to Figure 1 and Figure 2 , provides a microplate for primary cell 3D culture and functional evaluation test, including a microplate body and a plurality of sink holes 11 arrayed on the microplate body.

[0027] Specifically, the specific structure of the microplate body is: Figure 3 and Figure 4 The microplate body includes a rectangular frame 2 and a main board 1 fixed within the rectangular frame 2. The top of the rectangular frame 2 is provided with an end plate 21 fixed to the four sides of the main board 1. The bottom of the rectangular frame 2 is hollowed out, and the four inner sides of the rectangular frame 2 are fixedly connected to the four outer sides of the main board 1 through multiple ribs 23. The bottom of the rectangular frame 2 is provided with outwardly protruding ridges 22, and a positioning notch is provided on one corner of the rectangular frame 2.

[0028] refer to Figure 2 A plurality of countersunk holes 11 are arranged in a rectangular array on the mainboard 1. In this embodiment, the number of countersunk holes 11 is 384, and they are arranged in 16 rows and 24 columns. Each countersunk hole 11 is a square hole, and the side length of each countersunk hole 11 is 3 mm to 4 mm. Preferably, in this embodiment, the side length of each countersunk hole 11 is 3.7 mm.

[0029] refer to Figure 2 and Figure 3 A plurality of sink grooves 12 are provided on the bottom surface of each sink hole 11. Each sink groove 12 is in the shape of an inverted regular quadrangular pyramid, and the inclination angle of each side of each sink groove 12 is 45° to 75°.

[0030] In this embodiment, four grooves 12 are provided in each countersunk hole 11. The four grooves 12 are arranged in a square array within the countersunk hole 11 and cover the entire bottom of the groove 12, thereby maximizing the space of the countersunk hole 11. Each side of each groove 12 has an inclination angle of 45°, a side length of 1.8 mm, a depth of 0.9 mm, and a spacing of 0.1 mm between adjacent grooves 12.

[0031] In this protocol, the steps for drug sensitivity testing of primary tumor organoids are as follows:

[0032] (1) Sample pretreatment: After the tumor tissue is fully digested, Cell sieve, centrifuge to collect cells, observe the cell sediment, if there are more red blood cells, then need to split the red blood cells. Resuspend in culture medium, add matrigel, and mix thoroughly with the volume of cell suspension: matrigel = 1:2.

[0033] (2) Microplate inoculation: Add 4 The cell matrix gel suspension was centrifuged at 4°C and 500G for 5 minutes. At this time, the cells will gather at the bottom of the sink 12. The microplate body was placed in an incubator for 30 minutes to solidify the matrix gel. 100 culture medium.

[0034] (3) Tumor organoid generation: After 5-7 days, tumor organoids can be generated in the main body of the microplate, and the medium is changed every two days.

[0035] (4) Drug sensitivity testing: After the tumor organoids are formed, the corresponding drugs can be added for treatment. During this period, real-time monitoring and photography can be performed. After 24-48 hours, the image data and drug treatment results can be analyzed.

[0036] Although the specific embodiments of the utility model are described in detail in conjunction with the accompanying drawings, this should not be construed as limiting the scope of protection of this patent. Within the scope described by the claims, various modifications and variations that can be made by those skilled in the art without creative work still fall within the scope of protection of this patent.

Claims

1. A microplate for primary cell 3D culture and functional evaluation test, characterized in that: The invention comprises a microplate body and a plurality of sink holes (11) arranged in an array on the microplate body; a plurality of sink grooves (12) are provided on the bottom surface of each sink hole (11), and each sink groove (12) is in the shape of an inverted regular quadrangular pyramid.

2. The microplate according to claim 1, wherein A plurality of the countersunk holes (11) are arranged in a rectangular array.

3. The microplate according to claim 1, wherein Each of the countersunk holes (11) is a square hole.

4. The microplate according to claim 3, characterized in that Four sink grooves (12) are provided in each sink hole (11), and the four sink grooves (12) are arranged in a square array in the sink hole (11) and cover the entire bottom of the sink groove (12).

5. The microplate according to claim 1, wherein The side length of each countersunk hole (11) is 3 mm to 4 mm.

6. The microplate according to claim 1, characterized in that Each side surface of each of the sinks (12) has an inclination angle of 45° to 75°.

7. The microplate according to claim 1, wherein The microplate body comprises a rectangular frame (2) and a main board (1) fixed within the rectangular frame (2); an end plate (21) fixed to the four sides of the main board (1) is provided on the top of the rectangular frame (2).

8. The microplate according to claim 7, characterized in that The four inner side surfaces of the rectangular frame (2) are fixedly connected to the four outer side surfaces of the main board (1) via a plurality of ribs (23).

9. The microplate according to claim 7, characterized in that Outwardly protruding ridges (22) are provided around the bottom of the rectangular frame (2).

10. The microplate according to claim 7, characterized in that A positioning notch is provided on one corner of the rectangular frame (2).

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