Organ-like culture device and mold for manufacturing same

Abstract

The utility model relates to the technical field of organoid culture, and discloses an organoid culture device and a mold for manufacturing the organoid culture device, which comprise a culture device body, a liquid adding port, a loading chamber, a plurality of strip-shaped culture chambers and a plurality of annular culture chambers, the loading chamber is arranged in the middle of the culture device body, the liquid adding opening is formed in the loading chamber, the plurality of strip-shaped culture chambers radially extend out of the loading chamber, and the annular culture chamber is arranged on the culture device body and is communicated with the parts, far away from the loading chamber, of the plurality of strip-shaped culture chambers; according to the scheme, the situation that the growth rates of organoid cells in matrigel coagulated in the strip-shaped culture chambers are different easily can be effectively avoided; the organoid can absorb more nutrient substances, oxygen and other soluble factors, so that the sustainable growth time of the organoid is prolonged, and the survival rate of the organoid is increased.

Description

Technical Field

[0001] The utility model relates to the technical field of organoid culture, and more specifically, to an organoid culture device and a mold for manufacturing the organoid culture device. Background Art

[0002] Matrigel is used for 3D cell culture, which allows cells / organoids to grow in 3D space. During culture, organoid cells are mixed into the matrigel and then added to the organoid culture device. The matrigel is originally a liquid and will become a gel at room temperature. After the matrigel gels, the organoid culture device is removed and culture medium is added to carry out normal organoid culture. The matrigel is evenly distributed through the organoid culture device and is in uniform contact with the culture medium, so that nutrients are obtained evenly. The developing organoids embedded in the matrigel rely on passive diffusion to supply nutrients and remove waste. As the organoids grow and their metabolic requirements become higher and higher, the diffusion in the internal area of ​​the matrigel is limited, causing the viability of the organoids to gradually decrease, leading to core necrosis.

[0003] The invention patent application with application publication number CN116490603A discloses a device for culturing organoids, which may include: an entry hole configured to receive a solution; a loading chamber, wherein the entry hole is located in the loading chamber; and a plurality of culture chambers, wherein the culture chambers extend radially from the loading chamber so that the solution injected into the loading chamber via the entry hole is distributed into the plurality of culture chambers, wherein the plurality of culture chambers are open to the external environment and include protruding edges at the openings of the plurality of culture chambers.

[0004] The above scheme increases the diffusion degree of organoid cells in the matrix gel from the central area to the outer area by extending a plurality of strip culture chambers radiating from the loading chamber, thereby improving the viability of the organoids. The sustainable growth time of the organoids is increased to at least 21 days, and the survival rate is increased to at least 80%. However, the end of the strip culture chamber away from the central area is a blind end and is relatively narrow and long. The organoid cells in the matrix gel condensed in each strip culture chamber are relatively isolated, and different growth rates are prone to occur. The diffusion degree of the organoid cells in the matrix gel condensed in the strip culture chamber is limited, and the sustainable growth time and survival rate of the organoids cannot be further improved. Utility Model Content

[0005] The utility model aims to overcome at least one defect of the above-mentioned prior art and provide an organoid culture device for solving the technical problem in the prior art that the growth time and survival rate of the organoid cannot be further improved due to the limited diffusion degree of the organoid cells in the matrix gel condensed in the strip culture chamber.

[0006] The technical solution adopted by the utility model is an organoid culture device, comprising a culture device body, a liquid adding port, a loading chamber, an annular culture chamber and a plurality of strip culture chambers; the loading chamber is arranged in the middle of the culture device body, the liquid adding port is arranged on the loading chamber, a plurality of strip culture chambers extend radially from the loading chamber, and the annular culture chamber is arranged on the culture device body and connects the parts of the plurality of strip culture chambers away from the loading chamber.

[0007] This solution connects the parts of multiple strip culture chambers away from the loading chamber by setting up an annular culture chamber, so that the distance between the matrix gel condensed in the strip culture chambers no longer exists, eliminating the defect of relative isolation of organoid cells in the matrix gel, and effectively avoiding the situation where the organoid cells in the matrix gel condensed in each strip culture chamber are prone to different growth rates; the diffusion degree of the organoid cells in the matrix gel condensed in the strip culture chamber is increased, and the surface area of ​​the matrix gel in contact with the culture medium is increased, so that the organoids wrapped in the matrix gel can absorb more nutrients, oxygen and other soluble factors, thereby improving the sustainable growth time and survival rate of the organoids.

[0008] Furthermore, the loading chamber is circular, and the annular culture chamber is concentric with the loading chamber, which is equivalent to dividing the annular culture chamber equally into each strip culture chamber, so that the culture medium contacted by the organoid cells in the matrix gel after coagulation is relatively uniform, thereby making the activity and growth rate of the organoid cells relatively uniform.

[0009] Furthermore, the annular culture chambers are concentrically and evenly spaced in the culture device body. The annular culture chambers are evenly spaced and cross-connected with the strip culture chambers, which overcomes the defect of relative isolation of the organoid cells in the matrix gel condensed in each strip culture chamber, so that the inoculated organoid cells can be uniformly absorbed into the culture medium, and the activity and growth rate of the cultured organoid cells can be more uniform.

[0010] Furthermore, the width of the strip-shaped culture chamber is equal to the radial width of the annular culture chamber. This arrangement allows the organoid cells to be more evenly distributed in the culture medium, so that the organoid cells evenly absorb the culture medium to achieve even growth.

[0011] Furthermore, the culture device body is made of PDMS material. Polydimethylsiloxane, also known as PDMS or dimethylsiloxane, is a polymer widely used in microfluidic chip manufacturing and biological experiment molding. It is non-toxic and odorless, has high transparency, is heat-resistant, hydrophobic, has low surface tension, is physiologically inert, and has good chemical stability.

[0012] The present scheme also discloses a mold for manufacturing an organoid culture device, including mold 1 and mold 2, wherein the mold 1 is provided with a support ring 1, and the mold 2 is provided with a support ring 2; when the mold 1 and the mold 2 are buckled, the support ring 1 and the support ring 2 are connected. During pouring, the support ring 1 and the support ring 2 are connected to block the pouring material to form an annular culture chamber on the body of the organoid culture device, thereby eliminating the relative isolation of the organoid cells between the matrix gel condensed in the strip culture chamber, improving the diffusion degree of the organoid cells in the condensed matrix gel, and increasing the surface area of ​​the matrix gel in contact with the culture medium, so that the organoid can absorb more nutrients, oxygen and other soluble factors.

[0013] Furthermore, the mold 1 is provided with a central pillar 1, the mold 2 is provided with a boss, and the boss is provided with a central pillar 2 concentric therewith; when the mold 1 and the mold 2 are buckled, the central pillar 1 and the central pillar 2 are connected. During pouring, the central pillar 1 and the central pillar 2 are connected to block the pouring material in the mold to form a liquid filling port, and the material between the boss and the mold 1 forms a loading chamber.

[0014] Furthermore, the mold 1 is provided with a plurality of strip pillars 1 around the central pillar 1, and the mold 2 is provided with a plurality of strip pillars 2 connected to the boss around the central pillar 2, and the plurality of strip pillars 1 and the plurality of strip pillars 2 are provided in a one-to-one correspondence; when the mold 1 and the mold 2 are buckled, the strip pillars 1 are connected to the corresponding strip pillars 2. The strip pillars 1 are connected to the corresponding strip pillars 2, blocking the casting material in the mold to form a plurality of strip culture chambers.

[0015] Furthermore, the mold 1 is provided with a positioning notch, and the mold 2 is provided with a positioning convex column, and the cross-sectional shape of the positioning convex column is the same as the shape of the positioning notch; when the mold 1 and the mold 2 are buckled, the positioning notch and the positioning convex column are plugged. Multiple positioning notches and positioning convex columns can be provided, which are mainly used to position the mold 1 so that the mold 1 can be accurately buckled on the mold 2.

[0016] Furthermore, the culture device body is formed by mixing PDMS liquid and PDMS curing agent and pouring them into the fastened mold 1 and mold 2 for curing. The mold 1 and mold 2 are printed by a 3D printer, and the sizes of the mold 1 and mold 2 can be adjusted according to different test requirements. The structure is simple, easy to operate, and more convenient to manufacture and use.

[0017] Compared with the prior art, the beneficial effects of the utility model are as follows: the present scheme connects the parts of multiple strip culture chambers away from the loading chamber by setting up an annular culture chamber, so that the spacing between the matrix gel condensed in the strip culture chambers no longer exists, eliminating the defect of relative isolation of organoid cells in the matrix gel, and can effectively avoid the situation where the organoid cells in the matrix gel condensed in each strip culture chamber are prone to different growth rates; the diffusion degree of the organoid cells in the matrix gel condensed in the strip culture chamber is increased, and the surface area of ​​the matrix gel in contact with the culture medium is increased, so that the organoids can absorb more nutrients, oxygen and other soluble factors, thereby improving the sustainable growth time and survival rate of the organoids. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a schematic diagram of the overall structure of an organ culture device according to an embodiment of the present invention.

[0019] Figure 2 This is a schematic diagram of the mold structure for manufacturing an organoid culture device in Example 2 of the utility model.

[0020] In the figure: 1. culture device body, 2. liquid adding port, 3. loading chamber, 4. strip culture chamber, 5. annular culture chamber, 6. mold 1, 7. mold 2, 8. support ring 1, 9. support ring 2, 10. center pillar 1, 11. center pillar 2, 12. boss, 13. strip pillar 1, 14. strip pillar 2, 15. positioning notch, 16. positioning boss. DETAILED DESCRIPTION

[0021] The drawings of the present invention are only used for illustrative purposes and cannot be construed as limiting the present invention. In order to better illustrate the following embodiments, some parts of the drawings may be omitted, enlarged or reduced, and do not represent the size of the actual product; for those skilled in the art, it is understandable that some well-known structures and their descriptions in the drawings may be omitted.

[0022] Embodiment 1

[0023] like Figure 1 As shown, the present scheme discloses an organoid culture device, comprising a culture device body 1, a liquid adding port 2, a loading chamber 3, a plurality of strip culture chambers 4 and an annular culture chamber 5; the loading chamber 3 is arranged in the middle of the culture device body 1, the liquid adding port 2 is arranged on the loading chamber 3, a plurality of strip culture chambers 4 extend radially from the loading chamber 3, the annular culture chamber 5 is arranged on the culture device body 1 and connects the parts of the plurality of strip culture chambers 4 away from the loading chamber 3, and the parts away from the loading chamber 3 are other parts on the strip culture chamber 4 except the parts connected with the loading chamber 3.

[0024] In this solution, an annular culture chamber 5 is provided to connect the parts of multiple strip culture chambers 4 away from the loading chamber 3, so that the distance between the matrix gel condensed in the strip culture chamber 4 no longer exists, eliminating the defect of relative isolation of the organoid cells in the matrix gel, and can effectively avoid the situation where the organoid cells in the matrix gel condensed in each strip culture chamber 4 are prone to different growth rates; the diffusion degree of the organoid cells in the matrix gel condensed in the strip culture chamber 4 is increased, and the surface area of ​​the matrix gel in contact with the culture medium is increased, so that the organoid can absorb more nutrients, oxygen and other soluble factors, thereby improving the sustainable growth time and survival rate of the organoid.

[0025] The loading chamber 3 is circular, and the annular culture chamber 5 is concentric with the loading chamber 3. It is equivalent to that each strip culture chamber 4 divides the annular culture chamber 5 into equal parts, so that the culture medium contacted by the organoid cells in the matrix gel after coagulation is relatively uniform, so that the activity and growth rate of the organoid cells are relatively uniform. There are several annular culture chambers 5 concentrically and evenly spaced in the culture device body 1. Several annular culture chambers 5 are cross-connected with the strip culture chambers 4 at equal intervals, which overcomes the defect of relative isolation of the organoid cells in the coagulated matrix gel in each strip culture chamber 4. Several annular culture chambers 5 can further increase the diffusion degree of the organoid cells in the coagulated matrix gel. The equal spacing arrangement makes the diffusion degree of the organoid cells increase more evenly from the center to the outside, which can make the activity and growth rate of the cultured organoid cells more uniform.

[0026] The width of the strip culture chamber 4 is equal to the radial width of the annular culture chamber 5. This arrangement allows the organoid cells to be more evenly distributed in the culture medium, so that the organoid cells evenly absorb the culture medium to achieve uniform growth. The culture device body 1 is made of PDMS material; polydimethylsiloxane, also known as PDMS or dimethylsiloxane, is a polymer widely used in microfluidic chip manufacturing and biological experiment molding. It is non-toxic and odorless, has high transparency, is heat-resistant, hydrophobic, has low surface tension, is physiologically inert, and has good chemical stability.

[0027] Embodiment 2

[0028] like Figure 2As shown, this embodiment discloses a mold for manufacturing an organoid culture device as in the first embodiment, including a mold 1 6 and a mold 2 7, a support ring 1 8 is arranged on the mold 1 6, and a support ring 2 9 is arranged on the mold 2 7; when the mold 1 6 and the mold 2 7 are buckled, the support ring 1 8 and the support ring 2 9 are connected. During casting, the support ring 1 8 and the support ring 2 9 are connected to block the casting material to form an annular culture chamber 5 on the organoid culture device body 1, thereby making the relative isolation of the organoid cells in the condensed matrix gel in the strip culture chamber 4 no longer exist, the diffusion degree of the organoid cells in the condensed matrix gel is improved, and the surface area of ​​the matrix gel in contact with the culture medium is increased, so that the organoid can absorb more nutrients, oxygen and other soluble factors. The support ring 1 8 and the support ring 2 9 can be concentrically arranged on the mold 1 6 and the mold 2 7 respectively to realize the casting of multiple annular culture chambers 5, and the mold 1 6 and the mold 2 7 can be selected from high temperature resistant photosensitive resin.

[0029] The mold 1 6 is provided with a central pillar 10, the mold 2 7 is provided with a boss 12, and the boss 12 is provided with a central pillar 2 11 concentric therewith; when the mold 1 6 and the mold 2 7 are buckled, the central pillar 10 and the central pillar 2 11 are connected. During pouring, the central pillar 10 and the central pillar 2 11 are connected to block the pouring material in the mold to form a liquid filling port 2, and the material between the boss 12 and the mold 1 6 forms a loading chamber 3.

[0030] A plurality of strip pillars 13 are arranged around the central pillar 10 on the mold 1 6, and a plurality of strip pillars 14 are arranged around the central pillar 11 on the mold 2 7, all of which are connected to the boss 12. The plurality of strip pillars 13 and the plurality of strip pillars 14 are arranged one by one in correspondence; when the mold 1 6 and the mold 2 7 are buckled, the strip pillars 13 and the corresponding strip pillars 14 are connected. The strip pillars 13 and the corresponding strip pillars 14 are connected in the mold to block the casting material, so as to form a plurality of strip culture chambers 4.

[0031] A positioning notch 15 is provided on the mold 1 6, and a positioning convex column 16 is provided on the mold 2 7. The cross-sectional shape of the positioning convex column 16 is the same as the shape of the positioning notch 15. When the mold 1 6 and the mold 2 7 are buckled, the positioning notch 15 is plugged into the positioning convex column 16. A plurality of positioning notches 15 and positioning convex columns 16 can be provided, which are mainly used to position the mold 1 6 so that the mold 1 6 can be accurately buckled on the mold 2 7.

[0032] The culture device body 1 is formed by mixing PDMS liquid and PDMS curing agent and pouring them into the buckled mold 1 6 and mold 2 7 for curing. The PDMS curing agent can be PDMS184PDMS curing agent, which belongs to the prior art and will not be described in detail here; the mold 1 6 and mold 2 7 are printed by a 3D printer, and the size of the mold 1 6 and mold 2 7 can be adjusted according to different test requirements. It has a simple structure, is easy to operate, and is more convenient to manufacture and use.

[0033] When in use: fit mold 1 6 onto mold 2 7, then mix the PDMS liquid and PDMS curing agent and pour them into the fitted mold 1 6 and mold 2 7, and cure them at 65°C for 2 hours to form them, then open mold 1 6, peel off the molding material from mold 2 7, and you can get an organoid culture device made of PDMS; finally, place the prepared organoid culture device into a cell culture well plate of matching size for organoid culture.

[0034] Obviously, the above embodiments of the utility model are only examples for clearly explaining the technical solution of the utility model, and are not intended to limit the specific implementation methods of the utility model. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the claims of the utility model shall be included in the protection scope of the claims of the utility model.

Claims

1. An organoid culture device, characterized in that: The invention comprises a culture device body (1), a liquid adding port (2), a loading chamber (3), an annular culture chamber (5) and a plurality of strip culture chambers (4); the loading chamber (3) is arranged in the middle of the culture device body (1), the liquid adding port (2) is arranged on the loading chamber (3), a plurality of strip culture chambers (4) extend radially from the loading chamber (3), and the annular culture chamber (5) is arranged on the culture device body (1) and communicates with the portions of the plurality of strip culture chambers (4) away from the loading chamber (3).

2. The organoid culture device according to claim 1, characterized in that: The loading chamber (3) is circular, and the annular culture chamber (5) is concentric with the loading chamber (3).

3. The organoid culture device according to claim 2, characterized in that: A plurality of annular culture chambers (5) are concentrically and evenly spaced apart in the culture device body (1).

4. The organoid culture device according to claim 1, characterized in that: The width of the strip-shaped culture chamber (4) is equal to the radial width of the annular culture chamber (5).

5. An organoid culture device according to any one of claims 1 to 4, characterized in that: The culture device body (1) is made of PDMS material.

6. A mold for manufacturing the organoid culture device according to any one of claims 1 to 5, characterized in that: The invention comprises a mold 1 (6) and a mold 2 (7), wherein the mold 1 (6) is provided with a support ring 1 (8), and the mold 2 (7) is provided with a support ring 2 (9); when the mold 1 (6) and the mold 2 (7) are buckled together, the support ring 1 (8) and the support ring 2 (9) are in contact with each other.

7. A mold for manufacturing an organoid culture device according to claim 6, characterized in that: The mold one (6) is provided with a center pillar one (10), the mold two (7) is provided with a boss (12), and the boss (12) is provided with a center pillar two (11) concentric therewith; when the mold one (6) and the mold two (7) are buckled, the center pillar one (10) and the center pillar two (11) are in contact with each other.

8. The mold for manufacturing an organoid culture device according to claim 7, characterized in that: The mold 1 (6) is provided with a plurality of strip pillars 1 (13) around the central pillar 1 (10), and the mold 2 (7) is provided with a plurality of strip pillars 2 (14) connected to the bosses (12) around the central pillar 2 (11), and the plurality of strip pillars 1 (13) and the plurality of strip pillars 2 (14) are provided in a one-to-one correspondence; when the mold 1 (6) and the mold 2 (7) are buckled, the strip pillars 1 (13) are connected to the corresponding strip pillars 2 (14).

9. A mold for manufacturing an organoid culture device according to any one of claims 6 to 8, characterized in that: The mold 1 (6) is provided with a positioning notch (15), and the mold 2 (7) is provided with a positioning convex column (16). The cross-sectional shape of the positioning convex column (16) is the same as the shape of the positioning notch (15); the positioning notch (15) and the positioning convex column (16) are plug-fitted.

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