Bioreactor for dynamically pressurizing and culturing tubular organoid

Abstract

The utility model designs a bioreactor for dynamically pressurizing and culturing tubular organs. The bioreactor comprises a biological reaction bin, a bracket, a culture solution supply device and a pressurizing device, the biological reaction bin is arranged on the bracket; the biological reaction bin comprises a reaction container and an air bag; the air bag is arranged in the reaction container, and the reaction container is fixed on the bracket; the pressurizing device penetrates through the reaction container and is connected with the air bag; and the culture solution supply device is connected with the reaction container. The tubular organoid is placed between the air bag and the reaction container, the air bag is inflated and expanded so as to apply pressure to the tubular organoid to achieve dynamic culture, perfusion of culture liquid is achieved through the infusion tube and the peristaltic pump, and finally efficient, convenient and sterile dynamic expansion pressurization culture of the tubular organoid is achieved.

Description

Technical Field

[0001] The utility model belongs to the field of cell culture, and particularly relates to a bioreactor for dynamically pressurizing and culturing tubular organoids. Background Art

[0002] In the repair and reconstruction of tubular tissues and organs (such as trachea, esophagus, blood vessels, etc.), tissues from patients or artificial organoids are generally used. Artificial organoids generally include categories such as metals, ceramics, polymer materials, and biomaterial cell scaffolds. Among them, the repair expectation of biomaterial cell scaffolds is the best. However, the initial strength of the cell scaffold is not high, and it is easily damaged when directly implanted. It is necessary to use a bioreactor to culture it, and then use it for repair or reconstruction after it has a certain strength.

[0003] In order to simulate the in-vivo growth environment and promote cell growth, in addition to perfusion, it is necessary to pressurize and culture the organoids. Traditional pressurized culture is to apply hydrostatic pressure to the cell scaffold in a closed container. The environment simulated by this method is quite different from the growth environment of the tissue cells in the body. Existing mechanical pressurized culture adopts various schemes. The power systems used, the waveforms of the pressurization pressure changes, and other environmental conditions required for culture are all different. And those used to test the culture effects are mostly sheet-like organoids.

[0004] So far, no bioreactor that is direct, simple, low-cost, and can ensure dynamic expansion mechanical stimulation has been found. An ideal tubular organoid culture device should be able to ensure the nutrients and bioactive factors required for the growth of the culture, and the mechanical stimulation provided should also have the same frequency as the stimulation received in the body. The tubular organoid deforms under the mechanical force of divergent radiation. As the strength of the pressurized cultured tubular organoid increases, it is necessary to gradually increase the expansion pressure to finally realize the simulation of the mechanical environment of the culture. Summary of the Utility Model

[0005] In order to solve the problem that the existing tubular organoids cannot obtain a simulated strain environment similar to in-vivo pressurization, which leads to poor culture effects of tubular organoids, this application designs a bioreactor for dynamically pressurizing and culturing tubular organoids. By placing the tubular organoid between an airbag and a reaction container, the airbag inflates and expands to apply pressure to the tubular organoid to achieve dynamic culture, and perfusion of the culture solution is realized through an infusion tube and a peristaltic pump, finally realizing efficient, convenient, and aseptic dynamic expansion pressurized culture of tubular organoids.

[0006] A bioreactor for dynamically pressurizing and culturing tubular organoids includes a biological reaction chamber, a bracket, a culture solution supply device, and a pressurizing device;

[0007] The biological reaction chamber is arranged on the bracket;

[0008] The biological reaction chamber includes a reaction vessel and an airbag; the airbag is arranged inside the reaction vessel, and the reaction vessel is fixed on a bracket;

[0009] The bottom and top of the reaction vessel and the airbag are rigid for fixing purposes, and the rest are elastic.

[0010] Preferably, 3 to 5 support feet are distributed on the inner wall of the lower part of the reaction vessel to fix the airbag and prevent the culture from tilting and hitting the wall;

[0011] The pressurizing device is connected through the reaction vessel and the airbag;

[0012] The culture medium supply device is connected to the reaction vessel.

[0013] Preferably, the pressurizing device includes a three-way air pipe and an air pump;

[0014] The air pump is connected to the airbag through the three-way air pipe;

[0015] A barometric sensor is further arranged on the three-way air pipe.

[0016] Preferably, the three-way air pipe is further connected to an electric control valve. The first connection port of the three-way air pipe is connected to the air pump, the second connection port of the three-way air pipe is connected to the airbag, and the third connection port of the three-way air pipe is connected to the electric control valve.

[0017] Preferably, the culture medium supply device includes a peristaltic pump and a culture medium container;

[0018] A first infusion pipe is communicated between the top of the culture medium container and the reaction vessel. One end of a second infusion pipe is connected to the bottom of the reaction vessel, and the other end passes through the peristaltic pump and is connected to the culture medium container, that is, a peristaltic pump is arranged on the second infusion pipe to drive the flow of the culture medium.

[0019] Preferably, culture medium valves are further arranged on the first infusion pipe and the second infusion pipe to prevent the culture medium from flowing back by closing the reaction vessel during pressurization.

[0020] Preferably, the culture medium needs to exchange gas with the incubator, and a gas filtration membrane is further arranged on the culture medium container to filter external gas and prevent bacterial contamination.

[0021] The advantages and effects of this application are as follows:

[0022] The present application designs a bioreactor for dynamically pressurizing and culturing tubular organoids, which includes a biological reaction chamber, a bracket, a culture medium supply device, and a pressurizing device; the biological reaction chamber is arranged on the bracket; the biological reaction chamber includes a reaction container and an airbag; the airbag is arranged inside the reaction container, and the reaction container is fixed on the bracket; the pressurizing device passes through the reaction container and is connected to the airbag; the culture medium supply device is connected to the reaction container. By placing the tubular organoids between the airbag and the reaction container, the airbag inflates and presses on the tubular organoids to achieve dynamic culture, and the culture medium perfusion is realized through the infusion tube and the peristaltic pump, finally realizing the efficient, convenient, and aseptic dynamic expansion and pressurization culture of the tubular organoids. The present application develops a culture method of dynamic expansion and pressurization, which can better simulate the pressure changes borne by the tubular tissue in the human body and obtain better culture effects and mechanical strength.

[0023] The above description is only an overview of the technical solution of the present application. In order to be able to understand the technical means of the present application more clearly, so as to be implemented in accordance with the content of the specification, and in order to make the above and other purposes, features, and advantages of the present application more obvious and understandable, the following takes the preferred embodiments of the present application and combines with the drawings to describe in detail as follows.

[0024] According to the following detailed description of the specific embodiments of the present application in combination with the drawings, those skilled in the art will be more clear about the above and other purposes, advantages, and features of the present application. Description of the Drawings

[0025] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings required to be used in the description of the embodiments or the prior art. Obviously, the following drawings are some embodiments of the present application. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained according to these drawings. In all the drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, the elements or parts are not necessarily drawn to actual scale.

[0026] Figure 1 Design drawing of a bioreactor for dynamically pressurizing and culturing tubular organoids designed for the present application;

[0027] Reference numerals:

[0028] 1. Bracket; 101 Support foot; 2. Reaction container; 3. Airbag; 4. Three-way gas pipeline; 5. Air pump; 6. Pressure sensor; 7. Electric control valve; 8. Culture medium valve; 9. First infusion tube; 10. Culture medium container 1001 Filter membrane; 11. Second infusion tube; 12. Peristaltic pump. Detailed Description of the Embodiments

[0029] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the following will clearly and completely describe the technical solutions in the embodiments of this application with reference to the accompanying drawings in the embodiments of this application. Obviously, the described embodiments are part of the embodiments of this application, rather than all of them. In the following description, providing specific details such as specific configurations and components is only to help comprehensively understand the embodiments of this application. Therefore, those skilled in the art should clearly understand that various changes and modifications can be made to the embodiments described here without departing from the scope and spirit of this application. Additionally, for the sake of clarity and conciseness, descriptions of known functions and structures are omitted in the embodiments.

[0030] It should be understood that the "one embodiment" or "this embodiment" mentioned throughout the specification means that the specific features, structures, or characteristics related to the embodiment are included in at least one embodiment of this application. Therefore, the "one embodiment" or "this embodiment" that appears throughout the specification does not necessarily refer to the same embodiment. In addition, these specific features, structures, or characteristics can be combined in one or more embodiments in any suitable manner.

[0031] In addition, this application may repeat reference numerals and / or letters in different examples. This repetition is for the purpose of simplicity and clarity, and in itself does not indicate the relationship between the various embodiments and / or settings discussed.

[0032] The term "and / or" in this article is merely a description of the association relationship of associated objects, indicating that three relationships can exist. For example, A and / or B can represent: A exists alone, B exists alone, and both A and B exist simultaneously. The term " / and" in this article is a description of another association object relationship, indicating that two relationships can exist. For example, A / and B can represent: A exists alone, and both A and B exist. In addition, the character " / " in this article generally indicates that the associated objects before and after are in an "or" relationship.

[0033] The term "at least one" in this article is merely a description of the association relationship of associated objects, indicating that three relationships can exist. For example, at least one of A and B can represent: A exists alone, both A and B exist simultaneously, and B exists alone.

[0034] It should also be noted that in this article, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise", or any other variant thereof are intended to cover non-exclusive inclusion.

[0035] Embodiment 1

[0036] Please refer toFigure 1 , this embodiment mainly introduces a bioreactor for dynamically culturing tubular organoids, which includes a bioreaction chamber, a bracket 1, a culture medium supply device, and a pressurization device;

[0037] The bioreaction chamber is arranged on the bracket 1;

[0038] The bioreaction chamber includes a reaction container 2 and an airbag 3; the airbag 3 is arranged inside the reaction container 2 and fixed on the support feet 101 on the lower inner wall of the reaction container, and the reaction container 2 is fixed on the bracket 1;

[0039] The pressurization device is connected through the reaction container 2 and the airbag 3; the airbag can expand or contract with inflation and deflation, and the tubular organoid is placed in the gap between the airbag and the inner wall of the reaction container;

[0040] The culture medium supply device is connected to the reaction container 2.

[0041] Furthermore, the pressurization device includes a three-way air pipe 4 and an air pump 5;

[0042] The air pump 5 is connected to the airbag 3 through the three-way air pipe 4;

[0043] A pressure sensor 6 is also arranged on the three-way air pipe 4, and the pressure sensor 6 is used to monitor the air pressure.

[0044] Furthermore, the three-way air pipe 4 is also connected to an electric control valve 7. The first connection port of the three-way air pipe 4 is connected to the air pump 5, the second connection port of the three-way air pipe 4 is connected to the airbag 3, and the third connection port of the three-way air pipe 4 is connected to the electric control valve 7; the two ends of the first connection port and the second connection port are communicated;

[0045] The electric control valve 7 is externally connected to a control circuit for controlling the electric control valve 7.

[0046] Furthermore, the culture medium supply device includes a peristaltic pump 11 and a culture medium container 9;

[0047] A first infusion pipe 9 is communicated between the culture medium container 10 and the top of the reaction container 2. One end of the second infusion pipe 11 is connected to the bottom of the reaction container 2, and the other end passes through the peristaltic pump 12 and is connected to the culture medium container 10.

[0048] A culture medium valve 8 is also arranged on the first infusion pipe 9 and the second infusion pipe 11 to prevent the culture medium from flowing back by closing the reaction container 2 during pressurization.

[0049] The culture medium needs to exchange gas with the incubator. A gas filtration membrane 1001 is also arranged on the culture medium container 10 to filter external gas and prevent bacterial contamination.

[0050] The upper and lower ends of the reaction vessel 2 are connected to the infusion tube, and the culture medium is driven by a peristaltic pump to perfuse the tubular organoids in culture. When pressurization is required, the culture medium valve and the peristaltic pump are closed, the air pump and the control circuit are started, and the electric control valve 7 will open and close at a set frequency under the control of the control circuit. When the valve is closed, the air pump is turned on to inflate the airbag; when the valve is opened, the air pump is turned off and the airbag deflates. Through this cycle, the inflation and deflation of the airbag are realized, so as to dynamically expand and pressurize the tubular organoids in culture.

[0051] A bioreactor for dynamically pressurizing and culturing tubular organoids designed by the utility model provides pressure by an airbag, has a simple and reliable structure, low material cost, controllable applied pressure, and can obtain better culture effects.

[0052] The above are only the preferred embodiments of the utility model, and it does not limit the protection scope of the utility model. For those skilled in the art, the utility model can have various changes and modifications. All changes, modifications, substitutions, integrations, and parameter changes made to these embodiments by conventional substitutions or capable of realizing the same functions without departing from the principle and spirit of the utility model fall within the protection scope of the utility model.

Claims

1. A bioreactor for dynamically pressurizing and culturing tubular organoids, characterized in that: It comprises a biological reaction chamber, a support (1), a culture fluid supply device and a pressurizing device; The bioreactor chamber is arranged on a support (1); The biological reaction chamber comprises a reaction container (2) and an air bag (3); the air bag (3) is arranged in the reaction container (2), and the reaction container (2) is fixed on a support (1); The lower inner wall of the reaction container is provided with support feet (101); The pressurizing device passes through the reaction container (2) and is connected to the air bag (3); The culture solution supply device is connected to the reaction container (2).

2. A bioreactor for dynamically pressurizing and culturing tubular organoids according to claim 1, characterized in that: The pressurizing device comprises a three-way air delivery pipe (4) and an air pump (5); The air pump (5) is connected to the air bag (3) via a three-way air delivery pipe (4); The three-way gas transmission pipe (4) is also provided with an air pressure sensor (6).

3. A bioreactor for dynamically pressurizing and culturing tubular organoids according to claim 2, characterized in that: The three-way gas supply pipe (4) is also connected to the electric control valve (7), the first connection port of the three-way gas supply pipe (4) is connected to the air pump (5), the second connection port of the three-way gas supply pipe (4) is connected to the air bag (3), and the third connection port of the three-way gas supply pipe (4) is connected to the electric control valve (7).

4. The bioreactor for dynamically pressurizing and culturing tubular organoids according to claim 1, characterized in that: The culture solution supply device comprises a peristaltic pump (12) and a culture solution container (10); A first infusion tube (9) is connected between the culture liquid container (10) and the top of the reaction container (2); one end of the second infusion tube (11) is connected to the bottom of the reaction container (2), and the other end passes through a peristaltic pump (12) and is connected to the culture liquid container (10); The first infusion tube (9) and the second infusion tube (11) are also provided with a culture fluid valve (8); The culture liquid container (10) is also provided with a gas filter membrane (1001).

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