A culture plate for organoids

Abstract

The utility model provides a kind of organoid's culture plate, including plate body and at least one culture well being arranged on the plate body, the culture well has culture bottom surface;The culture bottom surface is equipped with the grid structure that is protruding upwards, the grid structure separates the culture bottom surface into multiple independent grid spaces;The grid space is configured to accommodate matrix glue, the height of the grid structure is set to be higher than the height of the contained matrix glue, organoid is effectively limited in respective grid space, so that researcher can repeatedly observe specific organoid under microscope.

Description

Technical Field

[0001] This utility model relates to the field of culture plate technology, and in particular to a culture plate for organoids. Background Technology

[0002] Organoids are miniature tissue structures that self-organize from stem cells or tissue-specific progenitor cells in a three-dimensional culture system, capable of mimicking the structure and function of corresponding organs in vitro. With the rapid development of organoid technology in disease modeling, drug screening, and personalized treatment, higher demands are being placed on the long-term stable culture, observation, and manipulation of organoids.

[0003] Currently, commonly used organoid cultures employ cell culture containers, such as standard multi-well plates like 6-well and 24-well plates. In this system, organoids typically grow in the culture medium in a spherical, suspended or semi-adherent form. However, the following drawbacks exist: The lack of physical constraints in the culture wells allows organoids to move or drift randomly during culture due to the flow of the culture medium. This makes it difficult to repeatedly locate specific target organoids under a microscope. Researchers must re-search for the target within the culture well area each time they observe. Utility Model Content

[0004] The purpose of this invention is to address the shortcomings of existing technologies by providing an organoid culture plate to solve the problem of difficulty in locating organoids during the culture process.

[0005] To achieve the above objectives, this utility model provides the following technical solution: An organoid culture plate includes a plate body and at least one culture well disposed on the plate body, the culture well having a culture bottom surface; The culture bottom surface is provided with an upwardly protruding grid structure, which divides the culture bottom surface into multiple independent grid-like spaces; The grid-like space is configured to accommodate the matrix adhesive, and the height of the grid structure is set to be higher than the height of the matrix adhesive it accommodates.

[0006] In a preferred embodiment, the barrier structure includes multiple partition bars arranged in a crisscross pattern.

[0007] In a preferred embodiment, the height of the partition strip is less than 1 mm.

[0008] In a preferred embodiment, the width of the partition strip is 100 µm–200 µm.

[0009] In a preferred embodiment, the planar projection of the lattice space is square, with a side length of 0.5mm-2mm.

[0010] In a preferred embodiment, a marking structure is provided on the culture bottom surface of the lattice space.

[0011] In a preferred embodiment, the plate and the barrier structure are made of a transparent polymer material.

[0012] Compared with existing technologies, this technical solution has the following advantages: By incorporating a grid-like barrier structure on the bottom surface of the culture wells, the culture bottom is divided into multiple independent grid spaces. This design fundamentally solves the problems of organoids being prone to drift and difficult to reposition in traditional flat-bottomed culture plates. Organoids are effectively confined within their respective grid spaces, ensuring positional stability throughout the culture cycle and enabling researchers to repeatedly observe specific organoids under a microscope. Attached Figure Description

[0013] Figure 1 This is a schematic diagram of the structure of the culture plate for the organoids described in this utility model.

[0014] In the figure: 100 plate body, 110 upper surface of plate body, 120 lower surface of plate body, 200 culture well, 210 culture bottom surface, 220 culture sidewall, 300 grid structure, 300a grid space, 310 partition strip, 311 transverse partition strip, 312 circular partition strip, 313 vertical partition strip. Detailed Implementation

[0015] The following description is intended to disclose the present invention so that those skilled in the art can implement it. The preferred embodiments described below are merely examples, and other obvious variations will occur to those skilled in the art. The basic principles of the present invention defined in the following description can be applied to other embodiments, modifications, improvements, equivalents, and other technical solutions that do not depart from the spirit and scope of the present invention.

[0016] Please refer to Figure 1 An embodiment of the present invention provides an organoid culture plate, including a plate body 100 and at least one culture well 200 disposed on the plate body, wherein the culture well 200 has a culture bottom surface 210; The culture bottom surface 210 is provided with an upwardly protruding grid structure 300, which divides the culture bottom surface 210 into multiple independent grid-like spaces 300a. The grid space 300a is configured to contain the matrix adhesive, and the height of the grid structure 300 is set to be higher than the height of the contained matrix adhesive.

[0017] Each of the grid spaces 300a is used to contain the matrix adhesive for embedding organoids. When the matrix adhesive is placed in the grid space 300a, the raised grid structure 300 can form a physical barrier to confine the organoids in their respective grid spaces 300a and prevent the organoids from shifting between different grid spaces 300a.

[0018] In addition, the height of the barrier structure 300 is set to be higher than the height of the matrix gel it contains, further preventing organoids in different grid spaces 300a from aggregating due to fluid flow disturbance during culture.

[0019] As can be seen, the design of the lattice space 300a provides an independent and fixed culture unit for each organoid. Researchers do not need to search for the target again within the entire culture well 200 each time they observe. They can quickly and accurately locate and track specific organoids over a long period of time based on the position coordinates of the lattice space 300a, which greatly improves the observation efficiency.

[0020] like Figure 1 As shown, the plate 100 includes an upper surface 110 and a lower surface 120 disposed opposite to each other. The lower surface 120 serves as a bearing surface for placement on a platform such as a desktop or microscope stage. The upper surface 110 faces upward, and at least one culture well 200 is formed on the upper surface 110. The vertical distance between the upper surface 110 and the lower surface 120 defines the thickness of the plate 100.

[0021] The culture well 200 includes a culture bottom surface 210 and a culture sidewall 220. The culture sidewall 220 extends downward from the upper surface 110 of the plate and is connected to the periphery of the culture bottom surface 210.

[0022] The vertical distance between the culture bottom surface 210 and the upper surface 110 of the plate defines the depth of the culture hole 200. The depth of the culture hole 200 is less than the thickness of the plate 100.

[0023] In this embodiment, the culture bottom surface 210 is circular, and correspondingly, the cross-sectional shape of the culture well 200 is also circular.

[0024] like Figure 1 As shown, the barrier structure 300 includes multiple partition bars 310, which are arranged in a crisscross pattern.

[0025] Specifically, the multiple partition bars 310 include a circular partition bar 312, multiple horizontal partition bars 311, and multiple vertical partition bars 313. The circular partition bar 312 is arranged near the periphery of the culture bottom surface 210. The multiple horizontal partition bars 311 and the multiple vertical partition bars 313 are connected perpendicularly to each other within the area enclosed by the circular partition bar 312, dividing the enclosed area into multiple arrayed grid spaces 300a.

[0026] Because the horizontal partition strip 311 and the vertical partition strip 313 are arranged perpendicularly, the projection of the grid space 300a onto the plane is square.

[0027] The side length of the lattice space 300a ranges from 0.5mm to 2mm. This size range provides a sufficient number of independent lattice spaces 300a within a single culture well 200, while ensuring that each lattice space 300a has adequate space for organoid growth. In a preferred embodiment, the side length is set to approximately 1mm.

[0028] refer to Figure 1 The width of the partition strip 310 is 100 µm–200 µm.

[0029] The height of the partition strip 310 is less than 1 mm. The preferred height is 200 µm–500 µm.

[0030] The height of the partition strip 310 is based on a key functional requirement: the height of the partition strip 310 must be greater than the height of the matrix gel contained within the grid space 300a. In practice, a pipette is used to spot the matrix gel suspension containing organoids into each of the grid spaces 300a in the form of microdroplets. The spotting volume must be controlled so that the droplets are contained within the grid space 300a, and the surface of the solidified matrix gel is lower than the top of the partition strip 310.

[0031] The plate 100 and the grid structure 300 are made of transparent polymer materials, such as polystyrene (PS) or cyclic olefin polymers (COP), to ensure excellent optical transparency and meet the requirements of high-resolution microscopy observation.

[0032] In terms of manufacturing process, the plate 100 and the grid structure 300 can be integrally formed. Specifically, they can be formed in one step through in-mold injection molding; or, the grid space 300a can be first processed on the polymer base plate through laser etching.

[0033] refer to Figure 1The culture bottom surface 210 of the lattice space 300a is provided with a marking structure. The marking structure can be etched numbers, letters, or other labels, and these markings provide a unique coordinate identifier for each lattice space 300a.

[0034] The method of using the organoid culture plate includes the following steps: S1. The organoids to be cultured are mixed evenly with liquid matrix gel at low temperature to form a cell-matrix mixed suspension. Then, the mixed suspension is spotted as microdroplets into each 300a grid space. After the matrix gel solidifies, an independent culture unit of the organoid is formed in each of the 300a grid spaces.

[0035] The matrix gel can be a basement membrane extract (such as Matrigel), collagen, sodium alginate, or other hydrogels that can form a three-dimensional scaffold.

[0036] S2. Add sufficient culture medium to culture well 200. The nutrients in the culture medium can penetrate the matrix gel through diffusion, providing support for the organoids embedded in the matrix gel.

[0037] The culture medium is a cell culture solution adapted to the type of organoid being cultured, and its specific components are known to those skilled in the art.

[0038] S3. Transfer the inoculated culture plate into a cell culture incubator and culture according to the standard conditions for the organoids being cultured. Microscopic observation can be used during culture. Researchers can repeatedly locate specific organoids based on the marker structure of the 300a lattice space.

[0039] S4. When changing the medium or treating the drug, the liquid can be gently added or aspirated along the culture sidewall 220 of the culture well 200 using a pipette. The operation method is the same as that of a conventional culture plate.

[0040] S5. After the experiment, the target organoids can be gently removed from the designated 300a grid space using a pipette or tweezers. For example, the culture plate can be pre-cooled to liquefy the matrix gel, and then gently pipetted to aspirate it. The removed organoids are used for subsequent staining, molecular biological analysis, etc.

[0041] The number of organoids contained in the matrix gel droplets formed within each 300a grid space can be adjusted according to experimental needs, allowing for both single organoid culture and multi-organoid cluster culture.

[0042] The embodiments described above are only used to illustrate the technical ideas and features of this utility model. Their purpose is to enable those skilled in the art to understand the content of this utility model and implement it accordingly. The scope of patent application of this utility model should not be limited by these embodiments. That is, any equivalent changes or modifications made in accordance with the spirit disclosed in this utility model still fall within the patent scope of this utility model.

Claims

1. A culture plate for organoids, characterized in that, It includes a plate body (100) and at least one culture well (200) provided on the plate body, the culture well (200) having a culture bottom surface (210); The culture bottom surface (210) is provided with an upwardly protruding grid structure (300), which divides the culture bottom surface (210) into multiple independent grid spaces (300a). The grid space (300a) is configured to contain the matrix adhesive, and the height of the grid structure (300) is set to be higher than the height of the contained matrix adhesive.

2. The organoid culture plate as described in claim 1, characterized in that, The barrier structure (300) includes multiple partition bars (310), which are arranged in a crisscross pattern.

3. The organoid culture plate as described in claim 2, characterized in that, The height of the partition strip (310) is less than 1 mm.

4. The organoid culture plate as described in claim 2, characterized in that, The width of the partition strip (310) is 100µm–200µm.

5. The organoid culture plate as described in claim 1, characterized in that, The planar projection of the lattice space (300a) is square, with a side length of 0.5mm-2mm.

6. The organoid culture plate as described in claim 1, characterized in that, The culture bottom surface (210) of the lattice space (300a) is provided with a marking structure.

7. The organoid culture plate as described in claim 1, characterized in that, The plate (100) and the barrier structure (300) are made of transparent polymer material.

Images