A cell culture device in an in vitro simulated pressure environment

Abstract

The present invention belongs to the field of biomedicine and discloses a cell culture device and method in an in vitro simulated pressing environment. It also provides a cell culture device and method for simulating pressing stimulation on cells planted on an elastic culture membrane. The cells planted on the elastic culture membrane can be simultaneously loaded with various massage and pressing stimulation effects as needed, and the force acting on the elastic culture chamber culture membrane is similar to that in the real environment, thereby achieving in vitro simulation of various pressing stimulations. The device can be used for research on local human rehabilitation and related research at the cell and gene level.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and discloses a cell culture device and method in an in vitro simulated pressing environment, and provides a device and technical solution for simulating the effect of pressing on the human body and realizing in vitro research on the pressing and massaging effects on the human body. Background Art

[0002] When cells in the human body are subjected to force, changes in their secretion occur. It has been proven that massage can accelerate the recovery of injured parts of the body. This may be related to changes in cell secretion after massage of the damaged part, leading to changes in the tissue microenvironment and accelerated blood circulation. The complex environment in the human body makes it difficult to quantify and parameterize the effects.

[0003] And we know that the effect of a single force on cells does not exist in reality. For this reason, we invented this device to realize simulated pressing in vitro, so that the state of cells can be directly reflected, thereby achieving a more intuitive analysis of the effects of different pressings on cells in tissues. Summary of the Invention

[0004] In order to realize a method of culturing cells under a simulated pressing state, the present invention provides an in vitro cell culturing device and method under a simulated pressing environment. The device realizes the effect of simulating a human body massage state in vitro.

[0005] The present invention comprises a simulated pressing assembly (1), a first peristaltic pump (2), a second peristaltic pump (3), a liquid storage bottle (4), and a buffer chamber (9) connected by a circulating liquid pipeline, which is filled with cell culture fluid (10) to form a culture fluid circulation system, and a simulated finger drive system (5) is used to simulate pressing on the back of the elastic cell culture membrane (1-3), and is controlled by a computer control system (6) through a computer control circuit (8) to achieve a simulated massage effect on the human body. The complete set of devices is placed in a CO2 incubator (11) to form a complete system; wherein the simulated finger drive system (5) is used to simulate pressing on the back of the elastic cell culture membrane (1-3). The pressing assembly (1) is composed of a bottom plate (1-1), a bearing block (1-2) fixed on the bottom plate (1-1), an elastic cell culture membrane (1-3) installed on the top, a simulation guide intermediate plate (1-4) installed on the elastic cell culture membrane (1-3), a simulation finger (1-7) placed in the simulation guide intermediate plate (1-4) and then covered with a packaging top plate (1-8), a first joint (1-5), a second joint (1-6), and a packaging top plate introduction control joint (1-9) are installed on the side of the bottom plate (1-1), and the assembly is sealed and assembled by fixing bolts (1-10).

[0006] The elastic cell culture membrane (1-3) described in the simulated pressing component (1) of the present invention has a thickness between 0.1 mm and 3 mm and is made of a highly elastic material such as silicone or rubber. Under 100% strain, the elastic cell culture membrane can still return to its original shape.

[0007] The simulated fingers (1-7) in the simulated pressing assembly (1) of the present invention can be spherical, elliptical, or other different shapes, and can be made of hard materials or soft rubber materials that are not easily worn or adhered to elastic cell culture membranes. The simulated fingers (1-7) are controlled by a simulated finger drive system (5) to achieve various motions of the simulated fingers (1-7). The motions can achieve different simulated massage modes to achieve up and down massage, and the drive mode can be hydraulic, pneumatic, or electric to achieve different up and down motions of the simulated fingers. There are at least four simulated fingers (1-7), and related motions or combined motions can be controlled individually.

[0008] The first peristaltic pump (2) and the second peristaltic pump (3) in the culture fluid circulation system of the present invention are used, but not limited to, to realize the exchange of culture fluid from the cell culture fluid in the simulated pressing component with the culture fluid in the liquid storage bottle (4); the liquid storage bottle has a filter (4-1) for gas exchange with the external environment, and the operation of the peristaltic pump can also be suspended to replace the culture fluid during long-term use; at the same time, the first peristaltic pump (2) and the second peristaltic pump (3) are connected to the simulated pressing component, so that the culture fluid in the liquid storage bottle (4) and the culture fluid in the simulated pressing component form a circulation effect, but the amount of culture fluid in the buffer chamber will not be increased or decreased.

[0009] In the present invention, the liquid storage bottle (4) is provided with a filter (4-1) to allow the gas in the liquid storage bottle to be exchanged with the outside world, and at the same time, required trace elements, drugs, etc. can be added to the culture fluid to act on the cultured cells.

[0010] The buffer chamber (9) of the present invention is connected to the culture chamber in the simulated pressing component and is filled with a flowing cell culture fluid (10). The liquid level (L) therein is higher than the height of the culture matrix portion in the simulated pressing component (1) and is connected to the atmosphere. When the simulated finger (1-7) is pressed, a space is formed to avoid the matrix in the simulated pressing component and the matrix flows into the buffer chamber (9).

[0011] The system of the present invention can be placed in an incubator with a constant temperature environment and suitable CO2 and O2 environments.

[0012] The driving simulation finger (1-7) in the present invention enables the elastic cell culture membrane or tissue planted with cells to achieve simulated pressing in a scheduled manner. This pressing can simulate different position states. By pressing the elastic cell culture membrane, it acts on the back side of the surface where the cells are planted, and the tissue deformation and displacement formed by the background support effect creates an effect similar to pressing on the human body.

[0013] When the system is running, the simulated fingers are used to perform pressing movements under different conditions within the set pressure range. The simulated finger drive system can independently control multiple simulated fingers, control the reciprocating motion speed of the simulated fingers, and the range of reciprocating motion. This controllable motion can be used to obtain the force distribution and strain state of the elastic cell culture membrane through computer calculation and analysis.

[0014] The beneficial effects of the cell culture device and method under simulated compression in vitro of the present invention are:

[0015] 1. The present invention enables cells planted in the elastic culture chamber to well simulate the forces and strains that cells are subjected to in a physiological environment;

[0016] 2. The present invention enables the study of co-stimulation of cells grown in elastic culture chambers under pressure, strain, and microcirculatory conditions in a highly reproducible engineering manner, enabling the study of cells and tissues under physiological and non-physiological, normal, and extreme conditions.

[0017] 3. The present invention can simulate the effects of Chinese medicine massage;

[0018] 4. The present invention aims to enable an elastic cell culture membrane or tissue implanted with cells to be stretched and reset in a desired manner while simultaneously achieving a change in the pressure of the environment in which the elastic cell culture membrane or tissue is located, thereby enabling the elastic cell culture membrane or tissue implanted with cells to achieve simulated pressing in a desired manner. This pressing can simulate different positional states. By pressing the elastic cell culture membrane, the back side of the elastic cell culture membrane implanted with cells is acted upon, and the tissue deformation and displacement formed by the background support effect creates an effect similar to that created by pressing on the human body.

[0019] Therefore, the present invention provides a cell culture device under a simulated compression state in vitro. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] 1. Simulated pressing device (1), first peristaltic pump (2), second peristaltic pump (3), liquid storage bottle (4), simulated finger drive system (5), computer control system (6), circulating liquid pipeline (7), computer control circuit (8), buffer chamber (9), cell culture fluid (10), CO2 incubator (11) 2. Simulated pressing assembly (1): bottom plate (1-1), bearing block (1-2), elastic cell culture membrane (1-3), simulated finger guide middle plate (1-4), first joint (1-5), second joint (1-6), simulated finger (1-7), packaging top plate (1-8), control joint (1-9), fixing bolt (1-10), atmospheric joint (9-1); elastic cell culture membrane simulated finger action surface (1-3.1), elastic cell culture membrane cell planting surface (13.2), elastic cell culture membrane limiting ring (1-3.3), filter (4-1).

[0021] Figure 1 Schematic diagram of a cell culture device system in an in vitro simulated pressure environment

[0022] Figure 2 Schematic diagram of a cell culture device system in an in vitro simulated pressure environment

[0023] Figure 3 A schematic diagram of an example of simulating a pressing device pressing

[0024] Figure 4 Schematic diagram of the original position of an example simulation pressing device

[0025] Figure 5 Schematic diagram of an example elastic cell culture membrane DETAILED DESCRIPTION

[0026] After the device through which the culture medium passes is fully sterilized, cells are planted on the elastic cell culture membrane.

[0027] The part in contact with the culture medium and connected with the sealing loop is sterilized and then dried.

[0028] Place all contents in a biosafety cabinet for assembly and connection.

[0029] Connect the entire system according to the system diagram and ensure that the culture medium does not leak;

[0030] Place the system in a CO2 incubator; computer controls the simulated finger drive system to achieve the set simulated finger movement mode;

[0031] Set the time and run the instrument.

[0032] When the instrument reaches the set running time and stops to complete the experiment, the sealed chamber is opened, the elastic culture chamber is removed, and the next experiment is carried out according to the experimental purpose.

[0033] The above description is a detailed description of the preferred embodiments of the present invention, but the embodiments are not intended to limit the scope of the patent application of the present invention. All equivalent changes or modifications completed under the technical solutions suggested by the present invention should fall within the scope of the patent covered by the present invention.

Claims

1. A cell culture device in an in vitro simulated pressure environment, characterized in that: The cell culture device comprises a simulated pressing component (1), a peristaltic pump 1 (2), a peristaltic pump 2 (3), a liquid storage bottle (4), and a buffer chamber (9); the device is connected by a circulating liquid pipeline and filled with cell culture liquid (10) to form a culture liquid circulation system, and the simulated finger drive system (5) is used to simulate pressing on the back of the elastic cell culture membrane (1-3), and the computer control system (6) is controlled through the computer control circuit (8) to simulate the massage effect on the human body. The complete device is placed in a CO2 incubator (11) to form a complete system; The simulated pressing assembly (1) is composed of a bottom plate (1-1), a fixed bearing block (1-2) on the bottom plate (1-1), an elastic cell culture membrane (1-3) installed on the elastic cell culture membrane (1-3), a simulated finger guide middle plate (1-4) installed on the elastic cell culture membrane (1-3), a simulated finger (1-7) placed in the simulated finger guide middle plate (1-4), and then covered with a packaging top plate (1-8), a first joint (1-5) and a second joint (1-6) are installed on the side of the bottom plate (1-1), the packaging top plate is connected to the control joint (1-9), and is sealed and assembled by fixing bolts (1-10).

2. The in vitro cell culture device under simulated pressure environment according to claim 1, characterized in that: The bearing blocks (1-2) in the simulated pressing assembly (1) can be made into different strip-shaped and spherical massage background supports according to the simulation of different pressing positions. The materials can be hard materials, cartilage-like materials, or soft materials that are biocompatible or non-toxic. The distance between the bearing blocks and the elastic cell culture membrane is between 3 and 50 mm.

3. The in vitro cell culture device under simulated pressure environment according to claim 1, characterized in that The elastic cell culture membrane (1-3) in the simulated pressing component (1) has a thickness between 0.1 mm and 3 mm and is made of silicone or highly elastic rubber. Under 100% strain, the elastic cell culture membrane can still return to its original shape.

4. The in vitro cell culture device under simulated pressure environment according to claim 1, characterized in that: The simulated fingers (1-7) in the simulated pressing assembly (1) can have spherical, elliptical, or other different shapes, and can be made of hard materials or soft rubber materials that are not easily worn or adhered to elastic cell culture membranes; the simulated fingers (1-7) are controlled by a simulated finger drive system (5) to realize various motions of the simulated fingers (1-7), and the motions can realize different simulated massage modes to realize up and down massage, and the driving mode can be hydraulic, pneumatic, or electric to realize different up and down motions of the simulated fingers; there are at least four simulated fingers (1-7), and related motions or combined motions can be controlled individually.

5. The in vitro cell culture device under simulated pressure environment according to claim 1, characterized in that: The first peristaltic pump (2) and the second peristaltic pump (3) in the culture fluid circulation system are used, but not limited to, to exchange the culture fluid from the cell culture fluid in the simulated pressing component with the culture fluid in the liquid storage bottle (4); the liquid storage bottle is provided with a filter (4-1) for gas exchange with the external environment, and the operation of the peristaltic pump can be suspended to replace the culture fluid during long-term use; at the same time, the first peristaltic pump (2) and the second peristaltic pump (3) are connected to the simulated pressing component, so that the culture fluid in the liquid storage bottle (4) and the culture fluid in the simulated pressing component form a circulation effect, but the amount of culture fluid in the buffer chamber will not be increased or decreased.

6. The in vitro cell culture device under simulated pressure environment according to claim 1, characterized in that The liquid storage bottle (4) is provided with a filter (4-1) to allow the gas in the liquid storage bottle to be exchanged with the outside world, and at the same time, required trace elements can be added to the culture solution, and the medicine acts on the cultured cells.

7. The in vitro cell culture device under simulated pressure environment according to claim 1, characterized in that The buffer chamber (9) is connected to the culture chamber in the simulated pressing component and is filled with a flowing cell culture fluid (10). The liquid level (L) therein is higher than the height of the culture matrix portion in the simulated pressing component (1) and is connected to the atmosphere. When the simulated finger (1-7) is pressed, a space is formed to avoid the matrix in the simulated pressing component and the matrix flows into the buffer chamber (9).

8. The in vitro cell culture device under simulated pressure environment according to claim 1, characterized in that The complete system can be placed under an incubator with a constant temperature environment and suitable CO2 and O2 environment.

9. The in vitro cell culture device under simulated pressure environment according to claim 1, characterized in that: The cell culture device drives the simulation fingers (1-7) to achieve simulated pressing of the elastic cell culture membrane or tissue planted with cells in a desired manner; this simulated pressing can simulate the deformation and displacement of the tissue formed by pressing the elastic cell culture membrane in different position states, acting on the back of the surface where the cells are planted, and having a background support effect, thereby forming an effect similar to pressing on the human body.

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