A microfluidic glomerular chip with intercellular signal transduction and filtration functions

Abstract

The present invention belongs to the technical field of human bionic organ chips, and in particular relates to a microfluidic glomerular chip with intercellular signal transduction and filtration functions, comprising: a valve control layer, an endothelial cell-mesangial cell culture layer, a podocyte culture layer and a support layer arranged in sequence from top to bottom; the endothelial cell-mesangial cell culture layer comprises an endothelial cell culture combination chamber and a mesangial cell culture combination chamber provided on a second substrate, the endothelial cell culture combination chamber and the mesangial cell culture combination chamber being connected via a channel; the valve control layer is used to control the on-off of the channel; the podocyte culture layer comprises a podocyte culture-primary urine test combination chamber provided on a third substrate, the liquid inlet end of the podocyte culture-primary urine test combination chamber being connected to the endothelial cell culture combination chamber via a porous membrane; the liquid inlet end of the endothelial cell culture combination chamber, the liquid outlet end of the mesangial cell culture combination chamber and the liquid outlet end of the podocyte culture-primary urine test combination chamber extending upward to the valve control layer.

Description

Technical Field

[0001] The present invention belongs to the technical field of human bionic organ chips, and in particular relates to a microfluidic glomerular chip with intercellular signal transduction and filtration functions. Background Art

[0002] The glomerulus is primarily composed of three cell types: mesangial cells, endothelial cells, and podocytes. During kidney development, podocytes produce vascular endothelial growth factor A, which recruits endothelial cells to migrate to the podocytes. These recruited endothelial cells then express platelet-derived growth factor BB, which attracts mesangial cells. Mesangial cells act as a "skeleton," separating the primitive capillary capsule into capillary loops, ultimately forming the glomerular structure.

[0003] After glomeruli are formed, complex signal transduction continues between cells. Endothelial cells and podocytes, both located on the capillary basement membrane, can conduct signaling through bidirectional diffusion of cytokine gradients. Endothelial cells are in direct contact with mesangial cells, allowing them to transmit signals through paracrine secretion and gap junctions. Abnormal expression of any intercellular signaling pathway can cause glomerular disease. Therefore, intervening in abnormal intercellular signaling may serve as a new target for the treatment of glomerular diseases and has broad application prospects in drug screening for glomerular diseases.

[0004] Conventional animal experiments and 2D cell cultures are unable to study intercellular signaling. Current 3D culture systems, such as microfluidic glomerular chips, typically only culture two cell types to simulate the function of the glomerular barrier, which does not conform to the actual cellular direction of blood flow in the human body. Therefore, a microfluidic glomerular chip with intercellular signaling and filtration capabilities is urgently needed to address this issue. Summary of the Invention

[0005] The purpose of the present invention is to provide a microfluidic glomerular chip with intercellular signal transduction and filtration functions to solve the above problems.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] A microfluidic glomerular chip with intercellular signal transduction and filtration functions, comprising: a model component, wherein the model component comprises a valve control layer, an endothelial cell-mesangial cell culture layer, a podocyte culture layer, and a support layer arranged in sequence from top to bottom;

[0008] The endothelial cell-mesangial cell culture layer includes a second substrate, an endothelial cell culture combination cavity and a mesangial cell culture combination cavity opened on the second substrate, and the endothelial cell culture combination cavity and the mesangial cell culture combination cavity are connected through a channel;

[0009] The valve control layer is used to control the on and off of the channel;

[0010] The podocyte culture layer includes a third substrate and a podocyte culture-original urine test combined cavity provided on the third substrate, wherein the liquid inlet end of the podocyte culture-original urine test combined cavity is connected to the endothelial cell culture combined cavity through a porous membrane;

[0011] The liquid inlet end of the endothelial cell culture combination chamber, the liquid outlet end of the mesangial cell culture combination chamber, and the liquid outlet end of the podocyte culture-primary urine test combination chamber extend through the valve control layer.

[0012] Preferably, there are multiple model components.

[0013] Preferably, the valve control layer includes a first substrate, a gas-filled chamber is provided in the middle of the bottom surface of the first substrate, a film is fixed in the gas-filled chamber, and a gas-filled hole communicating with the gas-filled chamber is provided on the top surface of the first substrate;

[0014] The inflation chamber is inflated with air to cause the film to expand and squeeze into the channel to control the opening and closing of the channel;

[0015] The first substrate is provided with a culture medium inlet channel for communicating with the liquid inlet end of the endothelial cell culture combination chamber, a mesangial cell culture medium outlet for communicating with the liquid outlet end of the mesangial cell culture combination chamber, and a podocyte culture medium outlet for communicating with the liquid outlet end of the podocyte culture-primary urine test combination chamber.

[0016] Preferably, the endothelial cell culture combination chamber includes an endothelial cell culture chamber, which is provided through the second substrate, one side of the endothelial cell culture chamber is connected to one end of the channel, the other side of the endothelial cell culture chamber is connected to one end of the first flow channel, and the other end of the first flow channel is connected to the culture medium inlet channel;

[0017] The bottom of the endothelial cell culture chamber is communicated with the podocyte culture-primary urine test combination chamber through the porous membrane.

[0018] Preferably, the mesangial cell culture combination chamber includes a mesangial cell culture chamber, which is opened on the second substrate, one side of the mesangial cell culture chamber is connected to the other end of the channel, the other side of the mesangial cell culture chamber is connected to one end of the second flow channel, and the other end of the second flow channel is connected to the mesangial cell culture medium outlet.

[0019] Preferably, the podocyte culture-primary urine test combined chamber includes a podocyte culture chamber, the podocyte culture chamber is provided on the third substrate, the shape of the podocyte culture chamber matches the endothelial cell culture chamber, the podocyte culture chamber is located directly below the endothelial cell culture chamber, and the top of the podocyte culture chamber is connected to the bottom of the endothelial cell culture chamber through the porous membrane;

[0020] One side of the podocyte culture chamber is connected to one end of a third flow channel, the other end of the third flow channel is connected to one end of the primary urinary cavity, the other end of the primary urinary cavity is connected to one end of a fourth flow channel, the other end of the fourth flow channel is connected to a liquid flow channel connection outlet, and the liquid flow channel connection outlet is connected to the podocyte culture medium outlet.

[0021] Preferably, the supporting layer includes a fourth substrate, and the fourth substrate is fixed to the bottom of the third substrate.

[0022] Preferably, the film is a PDMS film.

[0023] Preferably, the channel is a microchannel array, which is arranged on an arc-shaped partition between the endothelial cell culture chamber and the mesangial cell culture chamber. The microchannel array includes a plurality of microfluidic channels, one end of the microfluidic channel is connected to the endothelial cell culture chamber, and the other end of the microfluidic channel is connected to the mesangial cell culture chamber, and the distance between two adjacent microchannels is equal.

[0024] Compared with the prior art, the present invention has the following advantages and technical effects:

[0025] The present invention is provided with three cell culture chambers, namely, an endothelial cell culture combination chamber, a mesangial cell culture combination chamber and a podocyte culture-primary urine test combination chamber. The endothelial cell culture combination chamber and the podocyte culture-primary urine test combination chamber are separated by a porous membrane, allowing endothelial cells and podocytes to grow on the upper and lower sides of the porous membrane respectively, thereby achieving the purpose of simulating the barrier function of the glomerulus. The endothelial cell culture combination chamber is connected to the mesangial cell culture combination chamber through a channel, and the channel can be controlled by the valve control layer. The blood replacement culture medium enters the mesangial cell culture combination chamber through the endothelial cell culture combination chamber and the channel. By controlling the channel, the inflow and outflow of the blood replacement culture medium and the cell treatment fluid in the cell research process are controlled. A primary urine test chamber is provided in the podocyte culture-primary urine test combination chamber, and the blood replacement culture medium filtered through the glomerular barrier flows into the chamber. The present invention can culture glomerular endothelial cells, glomerular mesangial cells and podocytes in different culture chambers according to the structure of the glomerulus. The cells are connected by porous membranes or channels that can exchange information, so that the study of intercellular signal transduction can be carried out. The blood replacement culture medium enters the podocyte culture-original urine test combination chamber through the porous membrane through the endothelial cell culture combination chamber, and can also enter the mesangial cell culture combination chamber through the channel. It is more suitable for the real structure of the glomerulus than the previous dual-channel microfluidic organ-on-chip; the glomerular endothelial cells and podocytes grow on both sides of the porous membrane to form a glomerular barrier; the endothelial cell culture combination chamber and the mesangial cell culture combination chamber are connected by a channel, and the inflow and outflow of blood culture medium and cell treatment fluid are controlled by controlling the on-off of the channel, which facilitates the study of glomerular-related diseases and drug screening. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. Those skilled in the art can also derive other drawings based on these drawings without inventive work.

[0027] Figure 1 It is a schematic diagram of the structure of the present invention;

[0028] Figure 2 This is a bottom view of the first substrate of the present invention;

[0029] Figure 3 Schematic diagram of the second substrate structure of the present invention;

[0030] Figure 4 For the present invention Figure 3 A partial enlarged view of the middle part;

[0031] Figure 5 This is a schematic diagram of the third substrate structure of the present invention;

[0032] Among them, 1. first substrate; 2. second substrate; 3. third substrate; 4. fourth substrate; 5. endothelial cell culture combination chamber; 6. porous membrane; 7. podocyte culture-primary urine test combination chamber; 8. thin film; 9. channel; 10. mesangial cell culture combination chamber; 101. air-filled chamber; 102. air-filled hole; 103. culture medium inlet channel; 104. mesangial cell culture medium outlet; 105. podocyte culture medium outlet; 501. endothelial cell culture chamber; 502. first flow channel; 1001. mesangial cell culture chamber; 1002. second flow channel; 701. podocyte culture chamber; 702. third flow channel; 703. primary urine chamber; 704. fourth flow channel; 705. liquid flow channel connection outlet. DETAILED DESCRIPTION

[0033] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0034] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the present invention is further described in detail below with reference to the accompanying drawings and specific embodiments.

[0035] Reference Figures 1 to 5 The present invention discloses a microfluidic glomerular chip with intercellular signal transduction and filtration functions, comprising: a model component, the model component comprising a valve control layer, an endothelial cell-mesangial cell culture layer, a podocyte culture layer and a support layer arranged in sequence from top to bottom;

[0036] The endothelial cell-mesangial cell culture layer includes a second substrate 2, an endothelial cell culture combination chamber 5 and a mesangial cell culture combination chamber 10 provided on the second substrate 2, and the endothelial cell culture combination chamber 5 and the mesangial cell culture combination chamber 10 are connected via a channel 9;

[0037] The valve control layer is used to control the on and off of channel 9;

[0038] The podocyte culture layer includes a third substrate 3 and a podocyte culture-original urine test combined chamber 7 provided on the third substrate 3. The liquid inlet end of the podocyte culture-original urine test combined chamber 7 is connected to the endothelial cell culture combined chamber 5 through a porous membrane 6.

[0039] The liquid inlet end of the endothelial cell culture combination chamber 5, the liquid outlet end of the mesangial cell culture combination chamber 10 and the liquid outlet end of the podocyte culture-primary urine test combination chamber 7 extend through the valve control layer.

[0040] The present invention is provided with three cell culture chambers, namely, an endothelial cell culture combination chamber 5, a mesangial cell culture combination chamber 10 and a podocyte culture-primary urine test combination chamber 7. The endothelial cell culture combination chamber 5 and the podocyte culture-primary urine test combination chamber 7 are separated by a porous membrane 6, allowing the endothelial cells and podocytes to grow on the upper and lower sides of the porous membrane 6 respectively, thereby achieving the purpose of simulating the barrier function of the glomerulus. The endothelial cell culture combination chamber 5 is connected to the mesangial cell culture combination chamber 10 through a channel 9. The on-off of the channel 9 can be controlled by the valve control layer. The blood replacement culture medium enters the mesangial cell culture combination chamber 10 through the endothelial cell culture combination chamber 5 and the channel 9. By controlling the on-off of the channel 9, the inflow and outflow of the blood replacement culture medium and the cell treatment fluid in the cell research process are controlled. A primary urine test chamber is provided in the podocyte culture-primary urine test combination chamber 7, and the blood replacement culture medium filtered through the glomerular barrier flows into the chamber. The present invention can culture the endothelial cells, mesangial cells and podocytes of the glomerulus in different culture chambers according to the structure of the glomerulus. The cells are connected by a porous membrane 6 or a channel 9 that can exchange information, so that the study of intercellular signal transduction can be carried out. The blood replacement culture medium passes through the endothelial cell culture combination chamber 5 and the porous membrane 6 to enter the podocyte culture-original urine test combination chamber 7, and can also enter the mesangial cell culture combination chamber 10 through the channel 9. It is more suitable for the real structure of the glomerulus than the previous dual-channel microfluidic organ-on-chip; the glomerular endothelial cells and podocytes grow on both sides of the porous membrane 6 to form a glomerular barrier; the endothelial cell culture combination chamber 5 and the mesangial cell culture combination chamber 10 are connected by the channel 9, and the inflow and outflow of the blood culture medium and the cell treatment fluid are controlled by controlling the on-off of the channel 9, so as to facilitate the study of glomerular-related diseases and drug screening.

[0041] To further optimize the solution, multiple model components are provided.

[0042] One model component or multiple model components can be set on the same chip. When multiple model components are set on the same chip, parallel experiments can be carried out. The number of model components can be matched according to the number required for parallel experiments. The model components can be arranged in parallel or in a matrix. The present invention does not impose any restrictions on this.

[0043] The present invention preferably has four groups of model components, and the four groups of model components are symmetrically arranged in pairs on the same chip.

[0044] According to a further optimized solution, the valve control layer includes a first substrate 1, a gas-filled chamber 101 is provided in the middle of the bottom surface of the first substrate 1, a film 8 is fixed in the gas-filled chamber 101, and a gas-filled hole 102 is provided on the top surface of the first substrate 1 and communicates with the gas-filled chamber 101;

[0045] The air in the air-filled chamber 101 is inflated to expand the film 8 and squeeze it into the channel 9 to control the opening and closing of the channel 9;

[0046] The first substrate 1 is provided with a culture medium inlet channel 103 for communicating with the liquid inlet end of the endothelial cell culture combination chamber 5, a mesangial cell culture medium outlet 104 for communicating with the liquid outlet end of the mesangial cell culture combination chamber 10, and a podocyte culture medium outlet 105 for communicating with the liquid outlet end of the podocyte culture-primary urine test combination chamber 7.

[0047] According to a further optimized solution, the film 8 is a PDMS film.

[0048] Located at the top of the chip is the valve control layer, which includes a first substrate 1. The thickness of the first substrate 1 is preferably 1 mm. An air-filled chamber 101 is provided on the bottom surface of the first substrate 1. A film 8 is fixed in the air-filled chamber 101. The film 8 is selected to be a PDMS film. The PDMS film is fixed in the air-filled chamber 101 by bonding. The film 8 cooperates with the air-filled chamber 101 to form an air valve. The air-filled hole 102 is connected in the air-filled chamber 101. The film 8 is pressed down to contact or detach from the channel 9 by the presence or absence of gas pressure. The film 8 is ductile. When in contact with the channel 9, it can fill to the inside of the channel 9, thereby blocking the channel 9 to achieve the closure of the channel 9. When the channel 9 needs to be opened, the air-filled hole 102 is opened to relieve the pressure, or a negative pressure is applied to separate the film 8 from the channel 9, so that the chambers on both sides of the channel 9 can be connected.

[0049] The on-off setting of channel 9 can prevent cell crosstalk during cell digestion.

[0050] The size of the inflation chamber 101 is preferably 6 mm × 1.2 mm × 0.25 mm (length × width × height), the size of the film 8 is preferably 6 mm in length, 0.6 mm in width, and 100 um in thickness, and the inner diameter of the inflation hole 102 is 1 mm.

[0051] In a further optimized solution, the endothelial cell culture combination chamber 5 includes an endothelial cell culture chamber 501, which is disposed through the second substrate 2. One side of the endothelial cell culture chamber 501 is connected to one end of the channel 9, and the other side of the endothelial cell culture chamber 501 is connected to one end of the first flow channel 502, and the other end of the first flow channel 502 is connected to the culture medium inlet channel 103.

[0052] The bottom of the endothelial cell culture chamber 501 is connected to the podocyte culture-primary urine test combined chamber 7 through a porous membrane 6 .

[0053] The second layer of the chip is the endothelial cell and mesangial cell culture layer, which includes a second substrate 2. The thickness of the second substrate 2 is preferably 0.75 mm. An endothelial cell culture combination cavity 5 and a mesangial cell culture combination cavity 10 are opened on the second substrate 2. The endothelial cell culture combination cavity 5 and the mesangial cell culture combination cavity 10 are connected by a channel 9.

[0054] The endothelial cell culture combination chamber 5 includes an endothelial cell culture chamber 501 and a first flow channel 502. One end of the first flow channel 502 is connected to the culture medium inlet channel 103 for transporting culture medium and perfusing endothelial cell suspension. The other end of the first flow channel 502 is connected to the endothelial cell culture chamber 501. The bottom of the endothelial cell culture chamber 501 is connected to the podocyte culture-primary urine test combination chamber 7 through a porous membrane 6. The side of the endothelial cell culture chamber 501 away from the first flow channel 502 is connected to the mesangial cell culture combination chamber 10 through a channel 9.

[0055] A further optimized solution is provided, in which the mesangial cell culture combination chamber 10 includes a mesangial cell culture chamber 1001, which is opened on the second substrate 2, one side of the mesangial cell culture chamber 1001 is connected to the other end of the channel 9, and the other side of the mesangial cell culture chamber 1001 is connected to one end of the second flow channel 1002, and the other end of the second flow channel 1002 is connected to the mesangial cell culture medium outlet 104.

[0056] The mesangial cell culture combination chamber 10 includes a mesangial cell culture chamber 1001 and a second flow channel 1002. One side of the mesangial cell culture chamber 1001 is connected to the endothelial cell culture chamber 501 through the channel 9, and the other side is connected to one end of the second flow channel 1002. The other end of the second flow channel 1002 is connected to the mesangial cell culture medium outlet 104, which is responsible for the discharge of culture medium that has not been filtered by the glomerulus and the perfusion of mesangial cell suspension.

[0057] A further optimized solution is that channel 9 is a microchannel array, which is arranged on the arc-shaped partition between the endothelial cell culture chamber 501 and the mesangial cell culture chamber 1001. The microchannel array includes a plurality of microfluidic channels, one end of the microfluidic channel is connected to the endothelial cell culture chamber 501, and the other end of the microfluidic channel is connected to the mesangial cell culture chamber 1001, and the distance between two adjacent microchannels is equal.

[0058] The microchannel array consists of 10 microchannels, each of which has a size of 1000um×200um×200um (length×width×height), and adjacent microchannels are spaced 200um apart.

[0059] To conform to the physiological structure of blood vessels, the endothelial cell culture chamber 501 and the mesangial cell culture chamber 1001 are both circular or elliptical. The endothelial cell culture chamber 501 is composed of a semicircle with a radius of 5 mm and a semi-ellipse with a horizontal axis of 1 mm and a vertical axis of 10 mm, with a depth of 0.75 mm. When multiple endothelial cell culture chambers 501 are set in the same chip, the distance between adjacent endothelial cell culture chambers 501 is at least 1 mm.

[0060] The mesangial cell culture chamber 1001 is composed of a crescent shape formed by a parallel arc 1 mm away from the semi-elliptical arc of the endothelial cell culture chamber 501 and a semicircle with a radius of 5 mm and the midpoint of the line connecting the two ends of the arc as the origin, that is, the horizontal axis is 4 mm, the vertical axis is 10 mm, and the depth is 0.5 mm.

[0061] The size of the first flow channel 502 is preferably 5 mm×1 mm×0.5 mm.

[0062] The size of the second flow channel 1002 is preferably 5 mm×1 mm×0.25 mm.

[0063] The microchannel array is arranged corresponding to the film 8 .

[0064] Further optimizing the solution, the podocyte culture-primary urine test combined chamber 7 includes a podocyte culture chamber 701, which is provided on the third substrate 3. The shape of the podocyte culture chamber 701 matches that of the endothelial cell culture chamber 501. The podocyte culture chamber 701 is located directly below the endothelial cell culture chamber 501, and the top of the podocyte culture chamber 701 is connected to the bottom of the endothelial cell culture chamber 501 through the porous membrane 6.

[0065] One side of the podocyte culture chamber 701 is connected to one end of the third flow channel 702, the other end of the third flow channel 702 is connected to one end of the primary urinary cavity 703, the other end of the primary urinary cavity 703 is connected to one end of the fourth flow channel 704, and the other end of the fourth flow channel 704 is connected to the liquid flow channel connection outlet 705, which is connected to the podocyte culture medium outlet 105.

[0066] The third layer is a podocyte culture layer, which includes a third substrate 3. The thickness of the third substrate 3 is preferably 0.75 mm. A podocyte culture-primary urine test combination chamber 7 is provided on the third substrate 3. The podocyte culture-primary urine test combination chamber 7 includes a podocyte culture chamber 701, a third flow channel 702, a primary urine chamber 703, a fourth flow channel 704 and a liquid flow channel connection outlet 705.

[0067] Endothelial cell culture chamber 501 and podocyte culture chamber 701 are of equal size and arranged in a vertically aligned manner to simulate the glomerular filtration barrier. Podocyte culture chamber 701 is composed of a semicircle with a radius of 5 mm and a semi-ellipse with a horizontal axis of 1 mm and a vertical axis of 10 mm, with a depth of 0.4 mm.

[0068] The porous membrane 6 is 100 μm thick and is composed of a porous polyester membrane with four surfaces modified with basement membrane extract (BME) to facilitate the adhesion of podocytes. It is located between the endothelial cell culture chamber 501 and the podocyte culture chamber 701, acting as the basement membrane of the glomerulus. To prevent liquid leakage, the porous membrane 6 has a larger area than the endothelial cell culture chamber 501, and its length and width are preferably 10 mm and 6 mm respectively, with an effective pore size of 3 μm. The porous membrane 6 is used to filter the incoming liquid before it enters the podocyte culture chamber 701.

[0069] The original urinary cavity 703 is used to store filtered culture medium, simulating the function of Bowman's capsule. The size of the original urinary cavity 703 is 5×3×0.25 mm.

[0070] The third flow channel 702 and the fourth flow channel 704 have dimensions of 11 mm×1 mm×0.2 mm.

[0071] The liquid channel connection outlet 705 extends upward and penetrates the first substrate 1 , and is communicated with the podocyte culture medium outlet 105 , for discharging the filtrate filtered by the glomerulus.

[0072] In a further optimized solution, the supporting layer includes a fourth substrate 4 , which is fixed to the bottom of the third substrate 3 .

[0073] The upper and lower surface areas of the first substrate 1, the second substrate 2 and the third substrate 3 are equal, specifically selected to be 75 mm long and 25 mm wide. The fourth substrate 4 is a glass bottom plate, and its area is equal to or greater than the area of ​​the third substrate 3. In the present invention, it is specifically selected to be 75 mm long and 25 mm wide.

[0074] After the first substrate 1 , the second substrate 2 , the third substrate 3 and the fourth substrate 4 are fixed together, the size of the entire chip is approximately 75 mm×25 mm×4 mm (length×width×height).

[0075] In the description of the present invention, it should be understood that the terms "longitudinal", "transverse", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc., indicating the orientation or position relationship, are based on the orientation or position relationship shown in the accompanying drawings, and are only for the convenience of describing the present invention, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention.

[0076] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art should fall within the scope of protection defined by the claims of the present invention.

Claims

1. A microfluidic glomerular chip with intercellular signal transduction and filtration functions, characterized in that: include: A model component, comprising a valve control layer, an endothelial cell-mesangial cell culture layer, a podocyte culture layer, and a support layer arranged in sequence from top to bottom; The endothelial cell-mesangial cell culture layer comprises a second substrate (2), an endothelial cell culture combination chamber (5) and a mesangial cell culture combination chamber (10) provided on the second substrate (2), wherein the endothelial cell culture combination chamber (5) and the mesangial cell culture combination chamber (10) are connected via a channel (9); The valve control layer is used to control the opening and closing of the channel (9); The podocyte culture layer comprises a third substrate (3), a podocyte culture-primary urine test combined chamber (7) provided on the third substrate (3), and a liquid inlet end of the podocyte culture-primary urine test combined chamber (7) is connected to the endothelial cell culture combined chamber (5) via a porous membrane (6); The liquid inlet end of the endothelial cell culture combination chamber (5), the liquid outlet end of the mesangial cell culture combination chamber (10), and the liquid outlet end of the podocyte culture-primary urine test combination chamber (7) extend through the valve control layer.

2. The microfluidic glomerular chip with intercellular signal transduction and filtration functions according to claim 1, characterized in that: There are multiple model components.

3. The microfluidic glomerular chip with intercellular signal transduction and filtration functions according to claim 1, characterized in that: The valve control layer comprises a first substrate (1), a gas-filled chamber (101) is provided in the middle of the bottom surface of the first substrate (1), a film (8) is fixed in the gas-filled chamber (101), and a gas-filled hole (102) is provided on the top surface of the first substrate (1) and is communicated with the gas-filled chamber (101); The inflation chamber (101) is inflated with air to cause the film (8) to expand and squeeze into the channel (9) to control the opening and closing of the channel (9); The first substrate (1) is provided with a culture medium inlet channel (103) for communicating with the liquid inlet end of the endothelial cell culture combination chamber (5), a mesangial cell culture medium outlet (104) for communicating with the liquid outlet end of the mesangial cell culture combination chamber (10), and a podocyte culture medium outlet (105) for communicating with the liquid outlet end of the podocyte culture-primary urine test combination chamber (7).

4. The microfluidic glomerular chip with intercellular signal transduction and filtration functions according to claim 3, characterized in that: The endothelial cell culture combination chamber (5) comprises an endothelial cell culture chamber (501), the endothelial cell culture chamber (501) is arranged through the second substrate (2), one side of the endothelial cell culture chamber (501) is connected to one end of the channel (9), the other side of the endothelial cell culture chamber (501) is connected to one end of the first flow channel (502), and the other end of the first flow channel (502) is connected to the culture medium inlet channel (103); The bottom of the endothelial cell culture chamber (501) is connected to the podocyte culture-primary urine test combined chamber (7) through the porous membrane (6).

5. The microfluidic glomerular chip with intercellular signal transduction and filtration functions according to claim 4, characterized in that: The mesangial cell culture combination chamber (10) comprises a mesangial cell culture chamber (1001), wherein the mesangial cell culture chamber (1001) is provided on the second substrate (2), one side of the mesangial cell culture chamber (1001) is connected to the other end of the channel (9), the other side of the mesangial cell culture chamber (1001) is connected to one end of a second flow channel (1002), and the other end of the second flow channel (1002) is connected to the mesangial cell culture medium outlet (104).

6. The microfluidic glomerular chip with intercellular signal transduction and filtration functions according to claim 4, characterized in that: The podocyte culture-primary urine test combined chamber (7) comprises a podocyte culture chamber (701), the podocyte culture chamber (701) is provided on the third substrate (3), the shape of the podocyte culture chamber (701) matches the endothelial cell culture chamber (501), the podocyte culture chamber (701) is located directly below the endothelial cell culture chamber (501), and the top of the podocyte culture chamber (701) is connected to the bottom of the endothelial cell culture chamber (501) via the porous membrane (6); One side of the podocyte culture chamber (701) is connected to one end of a third flow channel (702), the other end of the third flow channel (702) is connected to one end of the primary urinary cavity (703), the other end of the primary urinary cavity (703) is connected to one end of a fourth flow channel (704), the other end of the fourth flow channel (704) is connected to a liquid flow channel connection outlet (705), and the liquid flow channel connection outlet (705) is connected to the podocyte culture medium outlet (105).

7. The microfluidic glomerular chip with intercellular signal transduction and filtration functions according to claim 1, characterized in that: The supporting layer comprises a fourth substrate (4), and the fourth substrate (4) is fixed to the bottom of the third substrate (3).

8. The microfluidic glomerular chip with intercellular signal transduction and filtration functions according to claim 3, characterized in that: The film (8) is a PDMS film.

9. The microfluidic glomerular chip with intercellular signal transduction and filtration functions according to claim 5, characterized in that: The channel (9) is a microchannel array, which is arranged on an arc-shaped partition between the endothelial cell culture chamber (501) and the mesangial cell culture chamber (1001). The microchannel array includes a plurality of microchannels, one end of each microchannel is connected to the endothelial cell culture chamber (501), and the other end of each microchannel is connected to the mesangial cell culture chamber (1001), and the distance between two adjacent microchannels is equal.

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