Cell 3D culture chip suitable for differential pressure environment

Abstract

The utility model provides a cell 3D culture chip suitable for pressure difference environment, which comprises a base and a plurality of culture tanks filled with culture solution, the culture tanks are arranged on the base, and separators are arranged in the culture tanks to divide the culture tanks into culture areas and variable pressure areas which are communicated with each other. And the top of the separator is provided with a vent hole for communicating the variable pressure area with the outside. According to the utility model, shear stress can be provided, so that an actual shear stress environment in the chip is closer to that of a human body, and the cell culture efficiency and culture quality are improved.

Description

TECHNICAL FIELD

[0001] The utility model relates to the field of biology technique, concretely relates to a cell 3D culture chip suitable for differential pressure environment. BACKGROUND

[0002] Cell 3D culture and organoid culture technology has important significance in researching cell behavior and disease mechanism. Cell 3D culture and organoid culture often need to reproduce various physical, chemical and biological factors that cells face in the body as much as possible in vitro to create an environment closer to the physiological state, so as to more accurately study the behavior, morphology, structure and function of cells. Cells in the body are affected by various physical factors, including pressure, tension and shear stress, so cell culture needs to simulate some more complex tissue or organ functions.

[0003] The human body environment simulation process provides shear stress for cell culture, which can be realized through the complex structure and fluid parameter control in the culture cavity to improve the growth and gene expression of cells, and the microenvironment created by the culture chip is closer to the microenvironment of cells in the human body than the traditional experimental environment, and is more easily to simulate the more complex microenvironment in the body.

[0004] The existing cell 3D culture chip has the problems of low cell culture efficiency, large difference between the shear stress provided by the microenvironment and the human body and poor gas circulation. UTILITY MODEL CONTENT

[0005] In view of the deficiencies of the prior art, the utility model provides a cell 3D culture chip suitable for differential pressure environment, which can provide shear stress to make the chip more close to the actual shear stress environment of the human body, improve the culture efficiency and culture quality of cells.

[0006] The technical scheme of the utility model is as follows:

[0007] A cell 3D culture chip suitable for differential pressure environment, comprising a base and a plurality of culture grooves filled with culture solution arranged on the base, a partition is arranged in the culture groove to divide the culture groove into a culture area and a pressure change area in communication with each other, and a ventilation hole for connecting the pressure change area with the outside is arranged on the top of the partition.

[0008] As a preferred embodiment, the base is in a stepped structure and is divided into a first base and a second base, and a chip cover is arranged on the base to cover the first base.

[0009] As a preferred embodiment, the gap between the chip cover and the first base A forms a gas cavity, and a ventilation gap for connecting the gas cavity with the outside is arranged on the bottom of the chip cover.

[0010] As a preferred embodiment, the partition comprises a partition cover arranged on the top of the culture tank and a hollow partition column arranged in the culture tank from the partition cover.

[0011] As a preferred embodiment, the partition is integrally formed with the base through the partition cover.

[0012] As a preferred embodiment, the partition is detachably connected with the base through the partition cover.

[0013] As a preferred embodiment, the liquid level of the culture solution is higher than the lowest point of the partition column in the culture tank, so that the culture area forms a closed space.

[0014] Compared with the prior art, the present application has the following advantages:

[0015] The utility model provides a cell 3D culture chip suitable for differential pressure environment, including base and a plurality of filling culture tank of culture solution which are set up on base, be provided with partition piece in culture tank and divide culture tank into culture area and pressure change area which communicate with each other, the top of partition piece is equipped with the vent hole of pressure change area with outside communication, can press down or raise the liquid level height in pressure change area 2B through changing the air pressure state in pressure change area 2B, thereby change the liquid level height in culture area 2A, in this way simulate the shear stress that cell is provided in human body environment body fluid, can make the shear stress environment in microenvironment more close to human actual shear stress environment, improve the culture efficiency and culture quality of cell. BRIEF DESCRIPTION OF DRAWINGS

[0016] In order to more clearly illustrate the technical scheme in the embodiment of the utility model or prior art, the following will briefly introduce the drawing needed to be used in embodiment or prior art description, obviously, the drawing in the following description only some embodiments of the utility model, for ordinary skilled person in the art, under the premise of not paying the creative labor, still can obtain other drawings according to these drawings.

[0017] Figure 1 It is a structure schematic view of the cell 3D culture chip suitable for differential pressure environment of the utility model;

[0018] Figure 2 It is a structure schematic view of the base in the cell 3D culture chip suitable for differential pressure environment of the utility model;

[0019] Figure 3 It is Figure 2 The cross section structure schematic view of;

[0020] Figure 4The utility model discloses a cross section structure schematic diagram of cell 3D culture chip suitable for differential pressure environment in the embodiment 2 of the utility model.

[0021] The drawings are identified as:

[0022] 1-base, 1A-first base, 1B-second base, 2-culture tank, 2A-culture area, 2B-pressure change area, 3-separator, 3A-separation cover, 3B-separation column, 31-vent hole, 4-chip cover, 41-vent gap. DETAILED DESCRIPTION

[0023] The technical scheme in the embodiments of the utility model will be clearly and completely described below with reference to the drawings in the embodiments of the utility model, and obviously, the described embodiments are only part of the embodiments of the utility model, not all the embodiments. Based on the embodiments in the utility model, all other embodiments obtained by the person skilled in the art without creative labor belong to the protection scope of the utility model.

[0024] In the description of the utility model, it needs to be explained that the orientation or position relation indicated by the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer" and the like is the orientation or position relation based on the drawing shown, and is only for the convenience of describing the utility model and simplifying the description, and therefore cannot be understood as the restriction on the utility model. In addition, the terms "first", "second", "third", "fourth" and the like are only for the description purpose, and cannot be understood as indicating or implying the relative importance.

[0025] In the description of the utility model, it needs to be explained that, unless otherwise explicitly specified and limited, the terms "mounting", "connection", "connecting" should be understood in a broad sense, for example, can be fixed connection, can also be detachable connection, or integrally connected, can be mechanical connection, can also be electrical connection, can be directly connected, can also be indirectly connected through the intermediate medium, and can be the communication inside two elements. For the person skilled in the art, the specific meaning of the above terms in the utility model can be understood according to the specific circumstances.

[0026] Before describing the embodiments of the utility model, it needs to be explained that cells are often exposed to fluid shear stress environment in vivo, such as vascular endothelial cells in blood are constantly subjected to the friction force of blood flow. Therefore, appropriate shear stress can be applied in the process of cell culture, which can simulate the physiological environment in vivo, so that the cells grow and differentiate under conditions closer to nature.

[0027] Specifically, appropriate shear stress can promote the growth and metabolism of cells. By stimulating cell surface receptors and signal transduction pathways, shear stress can activate intracellular metabolic pathways, improve cell energy supply and material metabolism levels, which helps cells maintain viability and proliferative capacity during culture, resulting in more cells for subsequent experiments or treatment. Therefore, it is particularly important to simulate the shear stress of fluid flow in the human body during cell culture. The following will specifically describe how to simulate the shear stress in the human body through a cell 3D culture chip to achieve the purpose of promoting cell culture.

[0028] Embodiment 1

[0029] Referring to Figures 1 to 3 The utility model discloses a cell 3D culture chip suitable for differential pressure environment, which can be applied to the fields of cell 3D culture, organoid culture and tissue culture, etc. The cell 3D culture chip comprises a base 1 and a plurality of culture tanks 2 filled with culture solution arranged on the base 1. A partition 3 is arranged in each culture tank 2 to divide the culture tank into a culture area 2A and a pressure change area 2B that are in communication with each other. The top of the partition 3 is provided with a vent hole 31 that connects the pressure change area 2B with the outside.

[0030] Specifically, each culture tank 2 on the base 1 is a cell culture group. The partition 3 divides the space in the culture tank 2 into the culture area 2A for cell culture and the pressure change area 2B for changing the liquid level in the culture tank 2 by injecting gas. The culture area 2A and the pressure change area 2B are in communication at the bottom of the culture tank 2, and the pressure change area 2B is in communication with the outside gas environment through the vent hole 31, so that the whole culture tank 2 can be connected with the outside air to ensure the gas circulation in the culture tank 2.

[0031] Further, the culture solution can flow into the culture tank 2 through the pressure change area 2B, and when the liquid level of the culture solution in the culture tank 2 exceeds the connecting position of the culture areas 2A and 2B at the bottom of the culture tank 2, the culture solution forms a closed environment in the culture area 2A. The liquid level in the pressure change area 2B can be lowered or raised by changing the air pressure state in the pressure change area 2B, so as to change the liquid level in the culture area 2A, thereby simulating the shear stress provided by the body fluid in the human body environment. When the air pressure in the pressure change area 2B is changed, a carbon dioxide incubator can be used to realize air pressure control, and at the same time, the PH value of the culture medium for cell culture can be maintained relatively stable. Specifically, the culture medium used for cell culture generally contains a buffer system such as sodium bicarbonate. When carbon dioxide is dissolved in the culture medium, it will react with water to generate carbonic acid, and the carbonic acid will dissociate into hydrogen ions and bicarbonate ions. During the metabolic process of cells, some acidic substances such as lactic acid will be produced, so that the pH value of the culture medium tends to decrease. The bicarbonate ions dissociated from the carbonic acid generated by the reaction of carbon dioxide and water can neutralize the acidic substances produced by cell metabolism, thereby offsetting the decrease in pH value to a certain extent and maintaining the relative stability of the pH value of the culture medium.

[0032] As a preferred embodiment, the base 1 is in a stepped structure and is divided into a first base 1A and a second base 1B. A chip cover 4 is arranged on the base 1 to cover the first base 1A. The chip cover 4 protects the culture tank 2 in the first base 1A, so that the culture tank 2 of the cell 3D culture chip is not damaged due to improper operation. The stepped structure design facilitates the operator to take the cell 3D culture chip through the second base 1B. The chip cover 4 supported on the first base 1A can also be taken along the edge of the second base 1B, reducing the operation difficulty of the operator.

[0033] As a preferred embodiment, the gap between the chip cover 4 and the first base 1A forms a gas cavity. A ventilation gap 41 is arranged at the bottom of the chip cover 4 to communicate the gas cavity with the outside. The cell 3D culture chip is generally placed in a culture box. The ventilation gap 41 can balance the air pressure in the culture box and the pressure change area 2B, and can change the liquid level in the culture area 2A by changing the air pressure in the culture box, so as to simulate the shear stress in the human body. The ventilation gap 41 supports the gas flow between the culture box and the pressure change area 2B. The carbon dioxide and other gases can enter the culture solution through the fluid cavity to maintain the relative balance of the acid-base in the culture environment.

[0034] As a preferred embodiment, the partition 3 comprises a partition cover 3A arranged on the top of the culture tank 2 and a hollow partition column 3B arranged in the culture tank 2 from the partition cover 3A, and the culture tank 2 is divided into a culture area 2A in the partition column 3B and a pressure change area 2B outside the partition column 3B by the hollow partition column 3B.

[0035] As a preferred embodiment, the partition 3 is integrally formed with the base 1 through the partition cover 3A, which can increase the stability and airtightness of the whole structure of the cell 3D culture chip, and ensure that the cell culture process in the culture tank 2 is not affected by the outside world. Furthermore, the material of the sealing cover can be soft silicone or other soft materials with elasticity, which can better seal the environment in the culture cavity 3.

[0036] As a preferred embodiment, the partition 3 is detachably connected with the base 1 through the partition cover 3A, and the partition cover 3A can also be detachably connected and arranged on the top of the culture tank 2, which can facilitate the observation and processing of the culture tank 2 by the operator.

[0037] As a preferred embodiment, the liquid level of the culture solution is higher than the lowest point of the partition column 3B in the culture tank 2, so as to form a closed space in the culture area 2A. The liquid level higher than the lowest point of the partition column 3B in the culture tank 2 can change the pressure in the culture area 2A and the pressure change area 2B respectively, the air pressure in the pressure change area 2B can be changed to adjust the liquid level height in the culture area 2A, and the change process of the liquid level difference can simulate the shear stress in the human body.

[0038] The working principle of the utility model is as follows: the cell 3D culture chip suitable for differential pressure environment is placed in a carbon dioxide incubator, the carbon dioxide concentration of the incubator changes the air pressure in the incubator, the pressure change area 2B in the cell 3D culture chip is connected with the environment of the incubator, the air pressure makes the culture solution in the pressure change area 2B move with the liquid level difference, so that the culture solution in the culture area 2A also moves up and down with the change of the liquid level height, the movement frequency can be controlled according to the frequency required by the simulation of human pulse and cell culture, so that an environment simulating the shear stress in the human body can be formed in the culture area 2A of the cell 3D culture chip, the shear stress environment in the microenvironment is closer to the actual shear stress environment of the human body, and the culture efficiency and culture quality of the cells are improved.

[0039] Example 2

[0040] Reference Figure 4The difference between the present embodiment 2 and embodiment 1 is that the bottom of the culture tank 2 is a hemispherical structure, wherein the hemispherical design helps to optimize the growth environment of cells in the 3D culture chip. On the one hand, the hemispherical curved surface can provide a more uniform cell attachment area, which is conducive to the uniform distribution and growth of cells. On the other hand, this design can also promote the flow and mixing of the culture medium in the chip, providing cells with more sufficient nutrients and oxygen while taking away metabolic waste, thereby maintaining a healthier cell growth environment.

[0041] Further, the hemispherical design can simulate the curved morphology of tissues or organs in the body to some extent, providing cells with a growth space closer to the in vivo environment. This design helps to study the changes in cell behavior in complex microenvironments, as well as the interactions between cells and cells, cells and matrix. The hemispherical design can also improve the culture efficiency of the cell 3D culture chip. Since the hemispherical curved surface can increase the cell attachment area and promote the flow and mixing of the culture medium, it can accelerate the growth and differentiation process of cells. At the same time, this design can also reduce the number of cells and the amount of culture medium required during the culture process, thereby reducing the cost of culture.

[0042] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A 3D cell culture chip suitable for differential pressure environments, characterized in that, It includes a base (1) and several culture tanks (2) filled with culture medium disposed on the base (1). The culture tank (2) is provided with a partition (3) to divide the culture tank into a culture area (2A) and a pressure-changing area (2B) that are interconnected. The top of the partition (3) is provided with a vent (31) that connects the pressure-changing area (2B) to the outside.

2. The cell 3D culture chip suitable for differential pressure environments according to claim 1, characterized in that, The base (1) has a stepped structure and is divided into a first base (1A) and a second base (1B). A chip cover (4) covering the first base (1A) is provided on the base (1).

3. The cell 3D culture chip suitable for differential pressure environments according to claim 2, characterized in that, The gap between the chip cover (4) and the first base (1A) forms a gas cavity, and the bottom edge of the chip cover (4) is provided with a ventilation gap (41) for connecting the gas cavity to the outside.

4. The cell 3D culture chip suitable for differential pressure environments according to claim 1, characterized in that, The separator (3) includes a separator cover (3A) disposed on the top of the culture tank (2) and a hollow separator column (3B) extending from the separator cover (3A) into the culture tank (2).

5. The cell 3D culture chip suitable for differential pressure environments according to claim 4, characterized in that, The separator (3) is integrally formed with the base (1) through the separator cover (3A).

6. The cell 3D culture chip suitable for differential pressure environments according to claim 4, characterized in that, The separator (3) is detachably connected to the base (1) via the separator cover (3A).

7. The cell 3D culture chip suitable for differential pressure environments according to claim 4, characterized in that, The liquid level of the culture medium is higher than the lowest point of the separator column (3B) located in the culture tank (2), so that the culture area (2A) forms a closed space.

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