CN103290145A - Pouring type bioreactor experiment method and device used by same - Google Patents

Abstract

The invention relates to the technical field of bioreactors in medical devices, and more specifically, to an experimental method for a perfusion bioreactor and a device for use thereof. The present invention proposes an experimental test method aimed at the dual influence of shear force and hydrostatic pressure caused by laminar flow on cells in a perfusion bioreactor, for the purpose of testing the culture medium under the action of gravity in the amplified culture of a perfusion bioreactor. The effect of static pressure applied to cells on cell culture provides a way to quantitatively study. The present invention designs a device that can quantitatively apply hydrostatic pressure on the set fluid shear force, and can provide cell culture under the condition of the set fluid shear force, and then provide the device that changes or remains constant according to the established law. Physical stimuli of varying hydrostatic pressure.

Description

A perfusion bioreactor experimental method and device for use thereof

technical field

The invention relates to the technical field of bioreactors in medical devices, and more specifically, to an experimental method for a perfusion bioreactor and a device for use thereof.

Background technique

Cell culture is the key to tissue engineering, and it is also the necessary scientific and technological basis for the formation of tissue engineering industry. The existing research on perfusion bioreactors for cell culture is based on the conditions of small-scale cell culture. When considering various physical force stimuli on cells, they are all based on the hydrostatic pressure being zero. Assumption, that is, under the premise that the acceleration of gravity of the liquid is zero, the fluid shear force of the fluid under the laminar flow model is mainly considered.

为流体流量，

为培养液的粘滞系数，b为图1中培养腔3的宽度，h为培养腔3的高度。

、b、h都是常量，只需调节培养液从培养液入口1进入培养腔3的流量，或调节培养液从培养液出口2流出的流量Q，就能获得所需的流体剪切力。 The existing main experimental tool for providing accurate fluid shear stress conditions under the laminar flow model is a parallel plate flow chamber device. Its structural diagram is shown in Figure 1, and its function is to cultivate The cells provide precise stimulation of fluid shear force, and study the effect of cell culture under the stimulation of different fluid shear force, in order to test the quantitative relationship between fluid shear force and cell culture effect. The principle is: in the parallel plate flow chamber shown in Figure 1, the values of length L and width b are much greater than the height h, then the fluid in the middle part of the culture chamber 3 can be considered as a fully developed laminar flow movement, and the fluid produced by the fluid The shear force can be calculated by Poiseuille's law as: ,

is the fluid flow,

is the viscosity coefficient of the culture solution, b is the width of the culture chamber 3 in FIG. 1 , and h is the height of the culture chamber 3 .

, b, and h are all constants, and the required fluid shear force can be obtained only by adjusting the flow rate of the culture solution entering the culture chamber 3 from the culture solution inlet 1, or adjusting the flow Q of the culture solution flowing out from the culture solution outlet 2.

However, in order to achieve large-scale cell culture, it is necessary to consider the problems caused by the scale-up of the existing small bioreactors. Due to the scale-up, the hydrostatic pressure generated by the liquid depth of the culture medium will affect the cell culture. No longer a physical-mechanical factor that can be ignored, it needs to be studied. That is, when the existing parallel plate flow chamber is used as an experimental method and tool for studying the mechanical stimulation factors produced by the culture medium in the perfusion bioreactor, it mainly has the following deficiencies:

(1) The existing research on the hydrodynamic effect in the perfusion bioreactor only considers the effect of the fluid shear force, and does not consider the effect of the hydrostatic pressure. In fact, when the bioreactor is scaled up, the hydrostatic pressure due to the liquid depth can no longer be ignored, and the influence of different hydrostatic pressures on cell culture needs to be considered;

(2) The existing experimental method for the influence of a single factor that can provide accurate hydrostatic pressure on cell culture is to place the cells in a culture tank with a certain liquid depth for static culture. Although this static culture method can Accurate hydrostatic pressure can be obtained by controlling the liquid depth, but the material exchange required by the cells will be difficult to proceed smoothly due to the non-flow of the culture medium, and the cell culture effect will be seriously affected due to the difficulty of material exchange, and the hydrostatic pressure cannot be tested correctly. factors contributing to stress;

(3) The existing parallel plate flow chamber can only provide accurate fluid shear force, but cannot provide accurate hydrostatic pressure at the same time.

(4) The existing experimental methods and devices for cell culture cannot provide accurate and controllable fluid shear force and accurate and controllable hydrostatic pressure at the same time, so that these two forces coexist independently of each other and can be adjusted independently force.

Contents of the invention

In order to overcome at least one of the defects described in the above-mentioned prior art, the present invention provides an experimental method for a perfusion bioreactor, which can provide accurate and controllable fluid shear force and accurate and controllable hydrostatic pressure at the same time, so that this The two forces are co-existing forces that do not affect each other and can be regulated separately. The cell culture can be studied on the influence of hydrostatic pressure on the cell culture under the condition of the flow culture of the culture fluid. Further, a device for using it is provided, which has a simple structure and is convenient to use.

In order to solve the above-mentioned technical problems, the technical solution adopted in the present invention is: a perfusion bioreactor experimental method, which comprises the following steps:

S1. According to the requirements of the shear force of the laminar flow fluid in the perfusion bioreactor, adjust the fluid flow at the inlet of the culture solution and the outlet of the culture solution at the same time, so that the shear force of the fluid in the parallel plate culture chamber is the same as that required in the perfusion bioreactor The shear force of the laminar flow fluid is consistent; wherein, the fluid flow regulation between the culture solution inlet and the culture solution outlet is realized by adjusting the inlet peristaltic pump connected to the culture solution inlet and the outlet peristaltic pump connected to the culture solution outlet.

S2. Keep the outflow flow rate of the culture solution at the outlet of the culture solution constant, increase the flow rate of the culture solution at the inlet of the culture solution, so that the culture solution enters the hydrostatic pipe, when the liquid level of the culture solution in the hydrostatic pipe rises to the required height h, restore the inflow of the culture solution at the culture solution inlet;

的液体静压力，

为培养液密度，g为重力加速度，h为液体静压管内液面的高度； S3. The liquid level of the culture medium in the hydrostatic pressure tube will be maintained at the required height h, so that the pressure in the parallel plate culture chamber can be provided as

hydrostatic pressure,

is the density of the culture medium, g is the acceleration of gravity, and h is the height of the liquid level in the hydrostatic tube;

S4. Keep the outflow flow rate of the culture solution at the outlet of the culture solution constant, adjust the inflow flow rate of the culture solution at the inlet of the culture solution, thereby adjusting the height h of the liquid level in the hydrostatic pressure tube, thereby adjusting the hydrostatic pressure in the parallel plate culture chamber the size of;

S5. Synchronously and incrementally adjust the inflow flow of the culture solution at the inlet of the culture solution and the outflow flow of the culture solution at the outlet of the culture solution, so as to adjust the magnitude of the fluid shear force in the parallel plate culture chamber while maintaining the hydrostatic pressure in the tube The liquid level remains unchanged.

Further, the synchronous incremental adjustment of the inflow and outflow fluids in the step S1 is specifically:

为流体流量，为培养液的粘滞系数，W为平行板培养腔的宽度，H为平行板培养腔的高度， 、W、H都是常量，只需调节培养液的流量Q，获得所需的流体剪切力。 When adjusting the inflow and outflow of the culture medium, ensure that the inflow and outflow flow are the same and adjusted synchronously, so that the liquid level h in the hydrostatic pressure tube is maintained at 0, and the fluid shear force generated by the fluid in the parallel plate culture chamber can be calculated according to Poiseuille's law is calculated as: ,

is the fluid flow, is the viscosity coefficient of the culture medium, W is the width of the parallel plate culture chamber, H is the height of the parallel plate culture chamber, , W, and H are all constants, and it is only necessary to adjust the flow Q of the culture medium to obtain the required fluid shear force.

Further, the liquid in the hydrostatic tube is relatively static, the liquid in the parallel plate culture chamber is a fully developed laminar flow movement, and static and dynamic liquids coexist, specifically:

The volume of the liquid entering the parallel plate culture chamber from the culture solution inlet is completely equal to the liquid volume flowing out through the culture solution outlet, so that the volume of the liquid in the parallel plate culture chamber remains unchanged, so that the liquid in the hydrostatic pressure tube can be kept constant. The liquid level remains constant;

提供精确的液体静压力，其中，

为培养液密度，g为重力加速度，h为液体静压管内液面的高度。 In the hydrostatic tube, the liquid at the bottom of the tube participates in the liquid flow in the parallel plate culture chamber, and the liquid above the bottom of the tube does not participate in the liquid flow in the parallel plate culture chamber. The hydrostatic pressure provided by the hydrostatic tube is regarded as The hydrostatic pressure in the static liquid state, so that the formula of the pressure force can be pressed

provide accurate hydrostatic pressure where,

is the density of the culture medium, g is the acceleration of gravity, and h is the height of the liquid level in the hydrostatic tube.

Further, the steps S4 and S5 realize the quantitative control of the fluid shear force and the hydrostatic pressure respectively, and can act on the experimental test method of cell culture at the same time, specifically:

Through step S4, the required hydrostatic pressure can be applied to the cell culture, and the hydrostatic pressure can be precisely quantitatively adjusted, can be changed according to a predetermined rule, and can also maintain a required constant value;

Through step S5, the required fluid shear force can be applied to the cell culture, and the fluid shear force can be precisely quantitatively adjusted, can be changed according to a predetermined rule, and can also be maintained at a required constant value.

A perfusion bioreactor experimental device, comprising a parallel plate culture chamber, a culture solution inlet and a culture solution outlet connected to the parallel plate culture chamber, wherein the parallel plate culture chamber is also provided with a hydrostatic tube, a hydrostatic tube The axial direction is perpendicular to the parallel plate culture chamber, and the hydrostatic pipe is connected with the parallel plate culture chamber; the parallel plate culture chamber is also connected with a liquid pressure measurement interface; the top of the hydrostatic pipe is provided with a filter membrane. The liquid pressure measurement interface can be connected with an inspection device so as to detect the liquid pressure in the parallel plate culture chamber.

Further, the parallel plate culture chamber includes a culture chamber bottom cover, a culture chamber upper cover arranged on the culture chamber bottom cover, and a culture chamber inner chamber arranged between the culture chamber bottom cover and the culture chamber upper cover; the culture chamber The bottom cover and the upper cover of the culture chamber are sealed and fixed by a sealing ring, and the culture solution inlet, culture solution outlet, and liquid pressure measurement interface are arranged on the outer wall of the culture chamber bottom cover, respectively communicating with the inner chamber of the culture chamber; The static pressure tube is arranged on the upper cover of the culture chamber, and its axial direction is perpendicular to the surface of the upper cover of the culture chamber, and communicates with the inner chamber of the culture chamber.

Further, the upper cover of the culture chamber is also provided with fixing screw holes, and the screws pass through the fixing screw holes to fix the upper cover of the culture chamber and the bottom cover of the culture chamber.

Specifically, the parallel plate culture cavity is a rectangular parallelepiped structure with a length of L=60mm, a width of W=20mm, and a height of H=2.5mm. The length L and width W of the parallel plate culture chamber are much greater than the value of its height. The filter membrane is a mesh membrane with a pore size of 0.2 microns. The filter membrane isolates external bacteria. The whole device is made of polymethacrylate Ester (PMMA) made.

Compared with the prior art, the beneficial effects are:

1) The present invention adopts an experimental method in which the accurately controllable fluid shear force and the accurately controllable hydrostatic pressure are independent of each other and can be regulated separately, so that strict and accurate regulation can be carried out according to their respective mechanical action formulas.

2) The present invention adopts the experimental method that the accurate and controllable fluid shear force and the accurate and controllable hydrostatic pressure can act on the same action point at the same time, so that the mechanical conditions of the cells at the action point and the effect of cell culture The relationship can be accurately quantified and analyzed, and the influence of a single factor of hydrostatic pressure can be separated, providing a quantifiable and accurate analysis method for the influence of hydrostatic pressure on cell culture in a perfusion bioreactor.

3) The experimental device adopted in the present invention can provide an implementation device for the experimental method proposed in the present invention, and can provide accurate and controllable fluid shear force and accurate and controllable hydrostatic pressure at the same time, and the two forces provided They can be independent of each other, can be regulated separately, and can act on the same point of action.

Description of drawings

Fig. 1 is a schematic diagram of the structure of an existing parallel plate flow chamber.

Fig. 2 is a schematic flow chart of the experimental method of the present invention.

Fig. 3 is a schematic diagram of the overall structure of the experimental device of the present invention.

Fig. 4 is an enlarged schematic diagram of the experimental device of the present invention.

Detailed ways

The accompanying drawings are for illustrative purposes only, and should not be construed as limitations on this patent; in order to better illustrate this embodiment, certain components in the accompanying drawings will 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 descriptions thereof may be omitted in the drawings.

As shown in Figures 3 and 4, a perfusion bioreactor experimental device includes a parallel plate culture chamber 40, a culture solution inlet 10 and a culture solution outlet 30 communicated with the parallel plate culture chamber 40, wherein the parallel plate culture chamber 40 Also be provided with hydrostatic pressure tube 50 on the top, the axial direction of hydrostatic pressure tube 50 is perpendicular to parallel plate culture cavity 40, hydrostatic pressure tube 50 is communicated with parallel plate culture cavity 40; Parallel plate culture cavity 40 is also connected with liquid pressure The measuring interface 20 ; the top of the hydrostatic pipe 50 is provided with a filter membrane 60 . The liquid pressure measurement interface 20 can be connected with an inspection device so as to detect the liquid pressure in the parallel plate culture chamber 40 .

The parallel plate culture chamber 40 comprises a culture chamber bottom cover 41, a culture chamber loam cake 43 arranged on the culture chamber bottom cover 41, a culture chamber inner chamber 42 arranged between the culture chamber bottom cover 41 and the culture chamber loam cake 43; The chamber bottom cover 41 and the culture chamber upper cover 43 are sealed and fixed by a sealing ring. The culture solution inlet 10, the culture solution outlet 30, and the liquid pressure measurement interface 20 are arranged on the outer wall of the culture chamber bottom cover 41, respectively communicating with the culture chamber inner chamber 42. ; The hydrostatic tube 50 is located on the upper cover 43 of the culture chamber, and its axial direction is perpendicular to the surface of the upper cover 43 of the culture chamber, and communicates with the inner chamber 42 of the culture chamber.

The culture chamber loam cake 43 is also provided with fixing screw holes 44, and the screws pass through the fixing screw holes 44 to fix the culture chamber loam cake 43 and the culture chamber bottom cover 41. The parallel plate culture chamber 40 is a rectangular parallelepiped, with a length of L=60mm, a width of W=20mm, and a height of H=2.5mm. The filter membrane 60 is a mesh membrane with a pore size of 0.2 microns. The length L and width W of the parallel plate culture chamber are much larger than the height, and its size meets the requirements of culturing cells. The filter membrane 60 isolates external bacteria, and the whole device is made of polymethyl methacrylate (PMMA). become.

，以在培养腔内室42内活动所需的流体剪切力：

，

为流体流量，

为培养液的粘滞系数，W为平行板培养腔40的宽度，H为平行板培养腔40的高度。在本发明提出的实验装置中，

、W、H都是常量，只需调节培养液的流量

，就能获得所需的流体剪切力。此时，由于培养液出、入口的流量始终保持一致，培养腔内室42内培养液的体积没有变化，使得培养腔内室42内注满培养液且液体静压管50内培养液的液面高度为0。 In the device shown in Figures 3 and 4, the method flow is shown in Figure 2, the glass slides planted with cells are placed in the central position of the inner chamber 42 of the culture chamber, and the culture chamber upper cover 43 is covered on the bottom of the culture chamber On the cover 41, there is an O-ring seal between them, and then it is reinforced from the fixing screw hole 44 with screws. Then only the peristaltic pump connected to the culture solution inlet 10 is started, and the culture solution is input from the storage tank into the culture chamber inner chamber 42. When the culture chamber inner chamber 42 is filled with the culture solution, the peristaltic pump connected to the culture solution outlet 30 is started. The pump adjusts its flow rate so that the culture chamber 42 is filled with culture solution and the hydrostatic pressure tube 50 is not filled with culture solution. Then adjust the two peristaltic pumps connected with the outlet and inlet of the culture solution synchronously and incrementally, so that the fluid flow in the inner chamber 42 of the culture chamber reaches the required flow value

, with the fluid shear force required for activities in the chamber 42 of the culture chamber:

,

is the fluid flow,

is the viscosity coefficient of the culture solution, W is the width of the parallel plate culture cavity 40, and H is the height of the parallel plate culture cavity 40. In the experimental device proposed by the present invention,

, W, H are all constant, just adjust the flow of the culture medium

, the desired fluid shear force can be obtained. At this time, because the flow rate of the culture solution out and in is always consistent, the volume of the culture solution in the culture chamber 42 does not change, so that the culture chamber 42 is filled with the culture solution and the liquid of the culture solution in the hydrostatic pressure tube 50 Face height is 0.

为培养液密度，g为重力加速度，h为液体静压管50内液面的高度。从而可为培养腔内室42提供压强为

的液体静压力。 Then keep the flow of the peristaltic pump connected to the culture solution outlet 30 constant, only increase the flow of the peristaltic pump connected to the culture solution inlet 10, so that the liquid surface in the hydrostatic pressure tube 50 rises to the required height h, and then Then the flow rate of the peristaltic pump connected to the culture solution inlet 10 is reduced to the flow rate of the peristaltic pump connected to the culture solution outlet 30, so that the liquid level of the culture solution in the hydrostatic pipe 50 remains constant at h. Due to the connection principle of liquid pressure, the liquid pressure received in the chamber 42 of the culture chamber is the same as the liquid pressure received at the bottom of the hydrostatic tube 50, both of which are provide accurate hydrostatic pressure where,

is the density of the culture solution, g is the acceleration of gravity, and h is the height of the liquid level in the hydrostatic tube 50 . Thereby, the pressure in the chamber 42 of the culture chamber can be provided as

of hydrostatic pressure.

，可通过调节培养腔内室42内培养液的流量调节其大小。 So far, it has been realized that two forces are provided at the cell culture place in the inner chamber 42 of the culture chamber, one of which is the hydrostatic pressure , its size can be adjusted by the height h of the liquid level in the hydrostatic tube; another force is the fluid shear force

, the flow rate of the culture solution in the chamber 42 of the culture chamber can be adjusted Adjust its size.

The pressure in the chamber 42 of the culture chamber can be observed in real time through the liquid pressure sensor connected to the liquid pressure measurement interface, so as to obtain accurate data of the hydrostatic pressure.

When the gas in the culture solution flows in the inner chamber 42 of the culture chamber, it can be released into the air from the filter membrane 60 in FIG. 3 , so that the pressure in the chamber 42 of the culture chamber will not be affected by the gas release. The filter membrane 60 is a mesh membrane with a pore size of 0.2 micron, which can pass gas and play the role of isolating bacteria.

The method for adjusting the size of h is as follows: keep the flow rate of the peristaltic pump connected to the culture solution outlet 30 constant, and only change the flow rate of the peristaltic pump connected to the culture solution inlet 10, so that the liquid surface in the hydrostatic pressure tube 50 rises to the required level. The required height h, and then the flow of the peristaltic pump connected to the culture solution inlet 10 is restored to the flow of the peristaltic pump connected to the culture solution outlet 30, so that the liquid level of the culture solution in the hydrostatic pipe 50 is maintained as desired The required h remains unchanged.

大小的方法是：同步同增量调节与培养液出、入口相连接的2个蠕动泵，使培养腔内室42内的流体流量达到所需的流量值

。 the regulation

The size method is: synchronously and incrementally adjust the two peristaltic pumps connected to the outlet and inlet of the culture solution, so that the fluid flow in the chamber 42 of the culture chamber reaches the required flow value

.

不变；所述的调节

的方法，在调节的同时，可以保持h不变。 The method for adjusting h, while adjusting h, can maintain

unchanged; adjustments as described

method, adjusting At the same time, h can be kept unchanged.

进行计算。 The hydrostatic pressure is a relatively static force. In the hydrostatic tube 50, only a small amount of culture fluid at the bottom will change due to the flow of the culture fluid in the chamber 42 of the culture chamber. Most of the above culture fluids are relatively static, and the hydrostatic pressure provided in the chamber 42 of the culture chamber can be strictly in accordance with

Calculation.

The experimental method proposed by the present invention enables cells to be cultured in a laminar flow environment while providing the effect of hydrostatic pressure, and overcomes the influence of unfavorable factors on the cell culture effect caused by material exchange when standing in a culture tank , providing a scientific and accurate quantitative experimental approach for evaluating the effect of hydrostatic pressure on cell culture in perfusion bioreactors.

Apparently, the above-mentioned embodiments of the present invention are only examples for clearly illustrating the present invention, rather than limiting the implementation of the present invention. For those of ordinary skill in the art, other changes or changes in different forms can be made on the basis of the above description. It is not necessary and impossible to exhaustively list all the implementation manners here. All modifications, equivalent replacements and improvements made within the spirit and principles of the present invention shall be included within the protection scope of the claims of the present invention.

Claims (9)

1. A perfusion bioreactor experimental method is characterized in that comprising the following steps: S1. According to the requirements of the shear force of the laminar flow fluid in the perfusion bioreactor, simultaneously adjust the fluid flow rate of the culture solution inlet (10) and the culture solution outlet (30), so that the shear force of the fluid in the parallel plate culture chamber (40) Consistent with the required laminar fluid shear in perfusion bioreactors; S2. Keep the outflow flow rate of the culture solution at the culture solution outlet (30) constant, increase the flow rate of the culture solution at the culture solution inlet (10), so that the culture solution enters the hydrostatic tube (50), when the hydrostatic tube (50 When the liquid level of the culture liquid in ) rises to the required height h, the inflow flow of the culture liquid at the culture liquid inlet (10) is resumed; S3. The liquid level of the culture solution in the hydrostatic pressure tube (50) will be maintained at the required height h, thus providing a pressure of hydrostatic pressure,

is the density of the culture solution, g is the acceleration of gravity, and h is the height of the liquid level in the hydrostatic tube (50); S4. Keep the outflow flow rate of the culture solution at the outlet of the culture solution (30) constant, and adjust the flow rate of the culture solution at the inlet of the culture solution (10), thereby adjusting the height h of the liquid level in the hydrostatic pressure tube (50), thereby adjusting the parallel The size of the hydrostatic pressure suffered in the plate culture cavity (40); S5. Synchronously and incrementally adjust the inflow flow rate of the culture solution at the culture solution inlet (10) and the outflow flow rate of the culture solution at the culture solution outlet (30), so that the height h of the liquid level in the hydrostatic pressure tube (50) remains unchanged, thereby Adjust the size of the fluid shear force in the parallel plate culture chamber (40). 2. The experimental method according to claim 1, characterized in that, in the step S1, the fluid flow of the culture solution inlet (10) and the culture solution outlet (30) is adjusted by adjusting the fluid flow rate connected to the culture solution inlet (10). The inlet peristaltic pump and the outlet peristaltic pump connected with the culture solution outlet (30) are realized. 3. experimental method according to claim 1, is characterized in that, in described step S1, to the synchronous increment adjustment of inflow and outflow fluid, be specially: When adjusting the inflow and outflow of the culture solution, ensure that the inflow and outflow flow are the same and adjusted synchronously, so that the liquid level h in the hydrostatic pressure tube (50) is maintained at 0, and the fluid generated in the parallel plate culture chamber (40) The fluid shear force can be calculated according to Poiseuille's law as:

,

is the fluid flow,

is the viscosity coefficient of the culture solution, W is the width of the parallel plate culture cavity (40), H is the height of the parallel plate culture cavity (40),

, W, and H are all constants, and it is only necessary to adjust the flow Q of the culture medium to obtain the required fluid shear force. 4. The experimental method according to claim 1, characterized in that, in the steps S2 and S3, the liquid in the hydrostatic tube (50) is relatively static, and the liquid in the parallel plate culture chamber (40) is fully developed The laminar flow movement, static and dynamic liquid coexist, specifically: The liquid volume entering the parallel plate culture chamber (40) from the culture solution inlet (10) is equal to the liquid volume flowing out through the culture solution outlet (30), so that the volume of the liquid in the parallel plate culture chamber (40) maintains constant, so that the liquid level in the hydrostatic pipe (50) remains constant; In the hydrostatic pressure tube (50), the liquid at the bottom of the tube participates in the liquid flow in the parallel plate culture chamber (40), and the liquid above the tube bottom does not participate in the liquid flow in the parallel plate culture chamber (40). The hydrostatic pressure provided by the tube (50) is regarded as the hydrostatic pressure in the static liquid state, so that the strong formula can be pressed

provide accurate hydrostatic pressure where, is the density of the culture solution, g is the acceleration of gravity, and h is the height of the liquid level in the hydrostatic tube (50). 5. An experimental device using any one of claims 1 to 4, comprising a parallel plate culture cavity (40), a culture solution inlet (10) communicated with the parallel plate culture cavity (40) and a culture solution outlet ( 30), characterized in that the parallel plate culture chamber (40) is also provided with a hydrostatic tube (50), the axial direction of the hydrostatic tube (50) is perpendicular to the parallel plate culture chamber (40), and the hydrostatic tube (50) communicates with the parallel plate culture cavity (40); the parallel plate culture cavity (40) is also connected with a liquid pressure measurement interface (20); the top of the hydrostatic tube (50) is provided with a filter membrane (60). 6. The experimental device according to claim 5, characterized in that, the parallel plate culture chamber (40) comprises a culture chamber bottom cover (41), a culture chamber upper cover arranged on the culture chamber bottom cover (41) (43), the culture chamber inner chamber (42) located between the culture chamber bottom cover (41) and the culture chamber upper cover (43); the culture chamber bottom cover (41) and the culture chamber upper cover (43) pass through the sealing ring Sealed and fixed, the culture solution inlet (10), culture solution outlet (30), and liquid pressure measurement interface (20) are arranged on the outer wall of the culture chamber bottom cover (41), and communicate with the culture chamber inner chamber (42) respectively The hydrostatic tube (50) is arranged on the upper cover (43) of the culture chamber, and its axial direction is perpendicular to the surface of the upper cover (43) of the culture chamber, and communicates with the inner chamber (42) of the culture chamber. 7. The experimental device according to claim 6, characterized in that, the upper cover (43) of the culture chamber is also provided with a fixing screw hole (44), and the screw passes through the fixing screw hole (44) to make the upper cover of the culture chamber Cover (43) is fixed with culture cavity bottom cover (41). 8. The experimental device according to claim 7, characterized in that the parallel plate culture chamber (40) is a cuboid structure with a length of L=60mm, a width of W=20mm, and a height of H=2.5mm. 9. The experimental device according to claim 8, characterized in that the filter membrane (60) is a mesh membrane with a pore size of 0.2 microns.

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