CN219279916U - Container for organoid culture - Google Patents

Abstract

The utility model provides a container for organoid culture, which comprises a container main body and an experiment groove positioned in the main body, wherein the experiment groove comprises a groove and a boss structure; the side of the groove is provided with a groove side wall, the boss structure is connected with the bottom surface of the groove, the boss structure is positioned in the groove and is provided with a horizontal top surface, the boss structure and the groove side wall of the groove are provided with intervals, and the height of the boss structure is smaller than that of the groove side wall.

Description

Container for organoid culture

Technical Field

The utility model relates to the technical field of organoid culture, in particular to a container for organoid culture.

Background

Currently, one of the key advances made in the field of stem cell research is organoid technology. As a tool, organoid technology has great potential in the study of a wide range of subjects, including developmental biology, disease pathology, cytobiology, regeneration mechanisms, precision medicine, and drug toxicity and efficacy testing.

Taking a tumor organoid as an example, the tumor organoid is taken from tumor tissue and is a miniature in vitro organ model obtained by culture and having the tissue characteristics of the source tumor and retaining the genetic characteristics of the primary tissue and the tumor heterogeneity. Tumor organoids are commonly used in disease model construction, mechanism studies, and screening for personalized precision medications for patients.

In addition, other organoids have also been studied with great importance in the respective fields and scenarios.

In order for organoids to function in the above scenario, it is critical to be able to achieve high-throughput, unified organoid culture.

Although the conventional organoid culture vessel has a plurality of experimental tanks, the conventional organoid culture vessel cannot culture organoids by a gas-liquid junction method.

Disclosure of Invention

Based on the problems, the utility model provides a container for organoid culture, which solves the problem that the existing multi-experiment tank container cannot culture organoids through a gas-liquid junction method in a mode of adding a boss structure into an experiment tank.

The embodiment of the utility model discloses a container which comprises the following components:

a container for organoid culture, said container comprising a container body and an experimental tank in the body, said experimental tank comprising a groove and a plateau formation; the side of recess is the recess lateral wall, boss structure with the recess bottom surface is connected, boss structure is located in the recess, boss structure has horizontal top surface, boss structure with the recess lateral wall of recess has the interval, boss structure's height is less than the height of recess lateral wall.

Optionally, the horizontal top surface of the container has a cover layer for tissue culture.

Optionally, the groove of the container is a cylindrical groove.

Optionally, the boss structure of the container is a cylindrical boss.

Optionally, the width of the horizontal top surface of the container is not less than the diameter of the smallest droplet of matrigel used for tissue culture.

Optionally, the container body has a plurality of the experiment slots.

Optionally, the plurality of experiment slots are arranged in an array on the container body.

Optionally, the width of the horizontal top surface of the boss structure is less than half the width of the groove.

Compared with the prior art, the utility model has the following beneficial effects:

according to the utility model, the boss structure is added in the experimental tank, and the matrigel can be placed on the horizontal top surface of the boss structure, so that the experimental tank can be applied to the culture of organoids by a gas-liquid junction method compared with the prior art.

In addition, the container disclosed by the utility model can comprise a plurality of experimental tanks, the culture environment in each experimental tank can be adjusted according to requirements, and the container provided by the utility model can be used for simultaneously carrying out experimental operation on organoids in a plurality of different culture environments. The experimental tanks can be arranged on the container main body according to an array, and can be compatible with various devices in the using process. The container disclosed by the utility model utilizes the boss structure to lift the organoid, so that the gas-liquid junction area of the organoid is increased, the preservation of the culture environment and the growth of the organoid can be considered on the premise of not influencing the nutrition supply of the organoid, and the characteristics of immunity and the like caused by the gas-liquid junction culture are preserved in the cultured organoid.

Drawings

In order to more clearly illustrate the embodiments of the utility model or the technical solutions of the prior art, the drawings which are used in the description of the embodiments or the prior art will be briefly described, it being obvious that the drawings in the description below are only some embodiments of the utility model, and that other drawings can be obtained according to these drawings without inventive faculty for a person skilled in the art.

FIG. 1 is a schematic illustration of a container provided by the present utility model;

FIG. 2 is a schematic diagram of an experimental tank provided by the utility model;

fig. 3 is a side cross-sectional view of an experimental tank provided by the utility model.

Detailed Description

As described above, the multi-well containers currently used in organoid culture do not support the gas-liquid interface method. If the culture in the culture dish is required by the gas-liquid interface rule, the process is complicated. The efficiency and throughput, i.e., throughput, of organoid culture in petri dishes is inadequate compared to multi-well containers. The culture dish often requires more culture fluid than a multi-well container, which is costly. When the gas-liquid junction method is used for testing the drug sensitivity of the organoids, the conventional multi-tank container for organoid culture does not support the gas-liquid junction method, and the efficiency and the output are insufficient when the culture is performed by using the culture dish.

In view of this, the present utility model proposes a container for organoid culture, specifically, the container can culture organoids by a gas-liquid interface method, the container having an experimental tank having a boss structure therein, the boss structure having a cover layer for tissue culture on a horizontal top surface thereof. When the gas-liquid junction method is used for testing the drug sensitivity of the organoids, the organoids can be lifted and suspended to be cultured through the boss structure, and a certain amount of culture solution is injected to test the drug sensitivity of the organoids through the gas-liquid junction method. The container provided by the utility model can be provided with a plurality of experimental tanks, and can be used for simultaneously carrying out sensitivity tests on multiple medicaments on the organoids, so that the efficiency and the output of organoid culture are improved compared with the existing container.

In order to make the present utility model better understood by those skilled in the art, the following description will clearly and completely describe the technical solutions in the embodiments of the present utility model with reference to the accompanying drawings, and it is apparent that the described embodiments are only some embodiments of the present utility model, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the utility model without making any inventive effort, are intended to be within the scope of the utility model.

Fig. 1 is a schematic view of a container provided by the present utility model.

The container schematic includes a container body 100 and an experiment slot 101 shown in phantom in the body, and the container schematic shown in fig. 1 includes numerals and letters on the container body for marking the experiment slot.

Fig. 2 is a schematic diagram of an experimental tank provided by the utility model.

Wherein, the experimental tank comprises a groove 201 and a boss structure 202. The boss structure is located in the groove, and the boss structure is spaced from the side wall of the groove. The grooves are cylindrical grooves, and the boss structure is a cylindrical boss.

Fig. 3 is a side cross-sectional view of an experimental tank provided by the utility model.

The boss structure 301 is connected with the groove bottom 303, the boss structure 301 has a horizontal top surface, the boss structure 301 and the groove side wall 303 have a certain interval, the boss structure has a horizontal top surface, and the height of the boss structure is smaller than that of the groove side wall.

In the case of the container shown in fig. 1, a plurality of test cells are provided, and a container having 96 test cells is used for illustration, but it is understood that other numbers of test cells are also possible and are within the scope of the present utility model. The center interval of two experimental slots of the universal 96-slot cell culture plate is 9 mm, and considering the experimental slot distribution of the universal 96-slot cell culture plate, the experimental slots in the container provided by the utility model can be consistent with the experimental slot distribution of the universal 96-slot cell culture plate, and can be consistent with the experimental slot distribution of other universal multi-slot cell culture plates, and the experimental slots can be distributed according to experimental requirements, which belongs to the protection scope of the utility model. The size of the experimental tank can be designed according to a common 96-tank cell culture plate or can be designed independently, and the experimental tank belongs to the protection scope of the utility model.

The container provided by the utility model can be compatible with the conventional devices such as an enzyme-labeled instrument, a living cell workstation, a laser confocal microscope and the like through the arrangement mode, can support other standard devices, and can realize the same functions of a universal 96-groove cell culture plate. The enzyme-labeled instrument, namely the enzyme-linked immunosorbent assay instrument, is a special instrument for enzyme-linked immunosorbent assay and is also called a microplate detector. The living cell workstation is an instrument for living cell imaging, cell migration determination, cell analysis optimization, cell culture quality control and cell proliferation analysis. The laser confocal microscope is an instrument for obtaining a fluorescence image of a microstructure inside a cell or a tissue by performing image processing by using a computer.

The container provided by the utility model is particularly suitable for organoids which have a volume of more than 500 mu m and are simultaneously required to be cultured by a gas-liquid junction method, and particularly comprises but is not limited to glioma, brain organs and the like.

In the experimental tank schematic diagram shown in fig. 2, a cylindrical groove and a cylindrical boss structure are selected as examples, however, the groove may be a groove with other shapes, and the boss structure may be a boss with other shapes, which are all in the protection scope of the present utility model.

In order to meet the experimental requirements, a culture solution can be injected into the groove, and the culture solution can be a tumor microenvironment required by a tumor organoid. The tumor microenvironment is composed of tumor cells, tumor-associated stromal cells, immune cells, and secretion products of the corresponding cells and non-cellular components of the extracellular matrix. The tumor microenvironment provides a good growth environment and nutrients for the tumor, and promotes the progress and metastasis of the tumor.

Wherein, the width of the horizontal top surface of the boss structure may be less than half the width of the groove. In practical application, the culture solution is placed in the groove, part of the space of the groove is formed by the boss structure, and the width of the horizontal top surface of the boss structure can be smaller than half of the width of the groove, so that the groove has enough space for placing the culture solution.

In the side cross-sectional view of the experimental tank shown in fig. 3, the horizontal top surface of the boss structure may have a cover layer for facilitating the attachment of the matrigel. Organoid culture typically requires placement of the organoid on a matrigel, and the coating on the horizontal top surface of the plateau provided by the present utility model provides room for the matrigel to perform organoid culture. In the practical experiment process, considering that the utility model needs to perform organoid culture on the horizontal top surface of the boss structure, the horizontal top surface of the boss structure is made of Tissue treated material, the horizontal top surface of the Tissue treated material is used as a covering layer, and the covering layer is convenient for the adhesion of matrigel, and can be a covering layer generated by other materials, and can be a covering layer without a covering layer, so that the utility model belongs to the protection scope of the utility model.

The matrigel is generally dripped on the position where organoid culture is needed through matrigel, and the width of the horizontal top surface of the boss structure is preferably not smaller than the diameter of the minimum liquid drop of matrigel in consideration of practical application scenes. Through the design, the matrigel can not drop into the groove, and the matrigel can be better attached to the horizontal top surface, so that the usability is stronger. Considering that the matrigel drop is generally one side attached to the horizontal plane and the other side is an ellipsoid after the horizontal plane, and considering the shape of the matrigel drop, the boss structure adopts a cylindrical boss, and the cylindrical boss is adopted to better adapt to experimental scenes.

In the scene that needs to cultivate organoids through the gas-liquid juncture method, pour into the culture solution into the recess, matrigel drops on the horizontal top surface of the boss structure, organoids are placed in matrigel, considering the demand of the gas-liquid juncture method, the organoids that cultivate need to contact with culture solution and air at the same time, here need to let the boss structure have a height smaller than the height of the recess lateral wall, meet the demand of the gas-liquid juncture method through such design.

In the scene of carrying out drug susceptibility testing on the organoids by utilizing a gas-liquid interface method, the organoids can be subjected to drug susceptibility testing in one experimental tank, and the container provided by the utility model can be used for simultaneously carrying out multiple drug susceptibility testing on the organoids due to the design of multiple experimental tanks. According to the actual test requirements, the types of the tested medicines are only required to be smaller than the number of the experiment grooves of the container, and compared with the medicine sensitivity test by using the culture dish, the medicine sensitivity test by using the container provided by the utility model has the advantages that the test efficiency and the output are improved.

The container provided by the utility model is provided with a plurality of experiment tanks, each experiment tank is used for culturing one organoid, when the cultured organoids are required to be subjected to drug susceptibility test, the drug susceptibility test can be directly carried out in the container provided by the application, and the organoids cultured by the existing container are required to be transferred from the culture container to the test container, so that a great amount of time is saved compared with the existing process.

Compared with the organoid cultured by the existing container, the organoid cultured by the container provided by the utility model can retain more immune cells in the organoid, and retain more immune cells so that the cultured organoid is more similar to the original tissue. Compared with the drug screening of the tumor antibody drug for the organoids cultured by the container, the drug screening of the tumor antibody drug for the organoids cultured by the container has more accurate results.

In a scene of culturing organoids by a gas-liquid interface method, preparing organoids, a culture solution with microenvironment and matrigel, placing the culture solution into a culture solution injection gun, and placing the matrigel into a matrigel gun. And (3) dripping the matrigel on the boss structure of the container provided by the utility model by using a matrigel gun, and performing certain treatment to enable the matrigel to be attached to the horizontal top surface of the boss structure. The organoids are placed on matrigel with the aid of a viewing instrument, such as a microscope. A quantity of culture fluid is injected into the well by means of a culture fluid injection gun such that the fluid level of the culture fluid contacts the matrigel but does not pass through the matrigel. Organoids are cultured, examined or observed.

In the present specification, each embodiment is described in a progressive manner, and each embodiment is mainly described in a different point from other embodiments, and identical and similar parts between the embodiments are all enough to refer to each other.

The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present utility model. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the utility model. Thus, the present utility model is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims (8)

1. A container for organoid culture, said container comprising a container body and an experimental tank in the body, said experimental tank comprising a groove and a plateau formation; the side of recess is the recess lateral wall, boss structure with the recess bottom surface is connected, boss structure is located in the recess, boss structure has horizontal top surface, boss structure with the recess lateral wall of recess has the interval, boss structure's height is less than the height of recess lateral wall.

2. The container of claim 1, wherein the horizontal top surface has a cover layer for tissue culture.

3. The container of claim 1, wherein the recess is a cylindrical recess.

4. The container of claim 1, wherein the boss structure is a cylindrical boss.

5. The container of claim 4, wherein the width of the horizontal top surface is no less than the diameter of the smallest droplet of matrigel used for tissue culture.

6. The container of claim 1, wherein the container body has a plurality of the assay channels.

7. The container of claim 6, wherein the plurality of test cells are arranged in an array on the container body.

8. The container of claim 1, wherein the width of the horizontal top surface of the boss structure is less than one-half the width of the groove.

Images