A cell co-culture system simulating hepatic sinusoids

Abstract

A cell co-culture system simulating hepatic sinusoids includes a multi-layer cell culture box, a microporous filter, a circulating peristaltic pump, a venous blood simulation box and an arterial blood simulation box. A culture fluid injection port is provided on the top of the culture box, an arterial blood interface and a venous blood interface are provided on the upper side wall of the culture box, and a liquid outlet is provided on the lower side wall of the culture box. The liquid outlet is connected to the venous blood simulation box and the arterial blood simulation box respectively through a circulation tube. The new culture system can realize the co-culture of multiple cells, simulate the real mechanical / chemical / biological microenvironment of cell growth by controlling the nutrient composition, flow rate, oxygen saturation and pH value of arteries and veins, and can perform drug screening experiments, study the interaction between different cells, analyze the conditions for tumor cell growth and the resulting environmental changes, and provide clues and basis for the prevention and treatment of liver diseases.

Description

Technical Field

[0001] The invention relates to a cell co-culture system for simulating liver sinusoids. Background Art

[0002] Traditional cell culture methods mostly grow in a flat surface in a culture bottle, while cells in the body are constantly affected by mechanical, chemical and biological factors in the body. The latest 3D cell culture system and co-culture system have solved some problems, but there is still a long way to go to build an in vitro cell culture system that simulates the real growth environment of cells in the body. The hepatic lobule is the basic unit for the liver to function. The hepatic sinusoids in the hepatic lobule mix arterial and venous blood, and the inner wall of the hepatic sinusoids is highly permeable. The hepatic sinusoid endothelial cells, liver / liver cancer cells and liver macrophages in the hepatic lobule have a unique mechanical / chemical / biological microenvironment, which cannot be simulated under ordinary in vitro culture. Therefore, it is urgent to build a multi-cell dynamic spatial co-culture system that simulates the hepatic sinusoid mechanical / chemical microenvironment to simulate the survival microenvironment of various cells in the hepatic lobule. It can also be combined and split to study the complex interactions between different cells in the hepatic lobule. Summary of the invention

[0003] In order to solve the deficiencies in the prior art, the present invention provides a multi-cell co-culture system that simulates the internal force / chemistry / biology microenvironment of liver sinusoids.

[0004] In order to solve the above technical problems, the present invention adopts the following technical scheme: a cell co-culture system simulating hepatic sinusoids, comprising a multi-layer culture box, a microporous filter, a circulating peristaltic pump, a venous blood simulation box and an arterial blood simulation box, a culture medium injection port is provided on the top of the multi-layer culture box, an arterial blood interface and a venous blood interface are provided on the upper side wall of the multi-layer culture box, a liquid outlet is provided on the lower side wall of the multi-layer culture box, and the liquid outlet is respectively connected to the venous blood simulation box and the arterial blood simulation box through a circulation tube, the microporous filter and the circulating peristaltic pump are sequentially arranged on the circulation tube, the venous blood simulation box is connected to the venous blood interface through a venous tube, the arterial blood simulation box is connected to the arterial blood interface through an arterial tube, a sample addition port is provided on the venous blood simulation box, and the arterial blood simulation box is also connected to an oxygen cylinder and a carbon dioxide cylinder.

[0005] The multi-layer incubator comprises a box body with an open front side, a box door is hinged at the open front side of the box body, a sealing strip is arranged on the periphery of the inner side of the box door, and an arterial blood interface and a venous blood interface are arranged on the box door; a first heating element is arranged inside the inner wall of the box body, a first display screen and a first microcontroller are arranged on the outer side of the box body, a first temperature sensor and a carbon dioxide concentration sensor are arranged on the inner wall of the box body, and the first heating element, the first temperature sensor, the carbon dioxide concentration sensor and the first display screen are respectively connected to the first microcontroller through lines;

[0006] A rectangular support member and two groups of wedge-shaped support members are fixed in the box, the rectangular support member is located between the two groups of wedge-shaped support members, the rectangular support member is composed of two long strip prisms, the two prisms are arranged side by side front and back, and the two ends of the prisms are respectively fixed on the left and right inner walls of the box, and the wedge-shaped support member is composed of two wedge-shaped support edges, the two support edges are arranged side by side front and back, and the two ends of the support edges are respectively fixed on the left and right inner walls of the box;

[0007] The top layer culture plate assembly is placed on the uppermost wedge-shaped support, the middle layer culture plate assembly is placed on the middle rectangular support, and the bottom layer culture plate assembly is placed on the lowermost wedge-shaped support;

[0008] The top culture plate assembly comprises a first rectangular frame, the bottom surface of the first rectangular frame is blocked by a bottom plate, the bottom plate is an inclined structure and is lower on the left and higher on the right, the inclination of the bottom plate is adapted to the inclination of the top surface of the supporting edge, a rectangular supporting inner frame is fixed to the inner frame wall of the first rectangular frame, a rectangular pressing frame body is arranged in the first rectangular frame, the pressing frame body is located above the supporting inner frame, the front side of the pressing frame body is hinged to the front side of the first rectangular frame through a pin shaft, a torsion spring is arranged on the pin shaft, a semipermeable membrane is arranged in the first rectangular frame, the periphery of the semipermeable membrane is arranged on the supporting inner frame of the first rectangular frame, and under the action of the torsion spring, the pressing frame body is pressed on the periphery of the semipermeable membrane;

[0009] The top culture plate assembly has the same structure as the bottom culture plate assembly; two first perforations are provided on the front side of the first rectangular frame of the top culture plate assembly; a venous blood inlet tube and an arterial blood inlet tube are placed on the upper surface of the semipermeable membrane; the venous blood inlet tube and the arterial blood inlet tube are placed side by side on the left and right; one end of the venous blood inlet tube passes through the first perforation and is connected to the venous blood interface of the box door; one end of the arterial blood inlet tube passes through another first perforation and is connected to the arterial blood interface of the box door; a plurality of first branches are connected to the venous blood inlet tube in parallel; a plurality of second branches are connected to the arterial blood inlet tube in parallel; a plurality of first branches and second branches are arranged one by one, and a plurality of liquid outlet holes are provided on the opposite surfaces of the first branches and the adjacent second branches; a plurality of first diversion interfaces are provided on the left side of the bottom plate;

[0010] A plurality of first through holes are provided on the right side frame of the first rectangular frame of the bottom culture plate assembly, the first through holes are in an L-shaped structure, a second flow guide interface is installed on the upper end of each first through hole, and a plurality of third flow guide interfaces are provided on the left side of the bottom plate of the bottom culture plate assembly, and the plurality of third flow guide interfaces are connected to the liquid outlet of the lower side wall of the multi-layer culture box through pipelines;

[0011] The middle culture plate assembly includes a second rectangular frame, the front frame wall and the rear frame wall of the second rectangular frame are both fixed with a long support plate, the rear frame wall of the second rectangular frame is provided with an arc groove, the arc groove is located above the long support plate, the right frame wall of the second rectangular frame is provided with a plurality of second through holes, the left frame wall of the second rectangular frame is provided with a plurality of third through holes, the second through holes and the third through holes are both L-shaped and located above the long support plate, the lower end of the second through hole is provided with a fourth flow guide interface, and the upper end of the third through hole is provided with a fifth flow guide interface;

[0012] The several first flow guide interfaces of the top culture plate assembly are respectively connected to the several fifth flow guide interfaces of the middle culture plate assembly through the flow guide hose, and the fourth flow guide interface of the middle culture plate assembly is respectively connected to the several second flow guide interfaces of the bottom culture plate assembly through the flow guide hose.

[0013] The venous blood simulation box comprises a box body, the inner cavity of the box body is divided into a solution cavity and a component installation cavity by a partition, a peristaltic pump is installed in the component installation cavity, a second display screen and a second microcontroller are arranged on the outer side of the box body, a pH sensor, a second temperature sensor and a blood oxygen saturation probe are arranged in the solution cavity, a second heating element is arranged on the inner wall at the bottom of the solution cavity of the box body, the peristaltic pump, the second display screen, the pH sensor, the blood oxygen saturation probe, the second temperature sensor and the second heating element are respectively connected to the second microcontroller through lines, and the liquid inlet pipe of the peristaltic pump is connected to the bottom of the solution cavity; a sample addition port is arranged at the top of the solution cavity of the venous blood simulation box;

[0014] The structure of the arterial blood simulation box is the same as that of the venous blood simulation box. The oxygen cylinder and the carbon dioxide cylinder are connected to the bottom of the solution chamber of the arterial blood simulation box through pipes, the circulation tube is connected to the upper part of the solution chamber of the arterial blood simulation box and the venous blood simulation box respectively, the outlet tube of the peristaltic pump of the arterial blood simulation box is connected to the arterial tube, and the outlet tube of the peristaltic pump of the venous blood simulation box is connected to the venous tube.

[0015] By adopting the above technical scheme, the present invention has the following beneficial effects: the present invention can realize multi-layer cell graded culture and simulate the growth environment of cells in the liver of a normal human body; the present invention can respectively extract the top culture plate assembly, the middle culture plate assembly and the bottom culture plate assembly to analyze the changes in the growth environment such as growth factors in different cell culture processes, which is beneficial to analyzing the growth characteristics and growth conditions of cells; the new technology can add anti-tumor drugs and other drugs at different levels to conduct experiments on drugs and other chemical substances. The new technology can control the flow rate, oxygen saturation and pH value of arteries and veins to further explore the conditions for cell growth, which is beneficial to analyzing the conditions for normal cell growth, and can conduct drug screening experiments to provide experimental data for anti-tumor drugs, etc. The conditions for tumor cell growth and the resulting environmental changes can be analyzed to provide clues for anti-tumor and provide a basis for treating diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 It is a schematic diagram of the structure of the present invention;

[0017] Figure 2 It is a schematic diagram of the structure of a multi-layer incubator;

[0018] Figure 3 is a schematic diagram of the structure of the top culture plate assembly;

[0019] Figure 4 It is an installation layout diagram of the semipermeable membrane, venous blood input tube and arterial blood input tube on the top culture plate assembly;

[0020] Figure 5 is a schematic diagram of the structure of the bottom culture plate assembly;

[0021] Figure 6 is a schematic diagram of the structure of the middle culture plate assembly;

[0022] Figure 7 It is a schematic diagram of the structure of the venous blood simulation box. DETAILED DESCRIPTION

[0023] like Figure 1-7 As shown, a cell co-culture system simulating hepatic sinusoids of the present invention comprises a multi-layer culture box 1, a microporous filter 2, a circulating peristaltic pump 3, a venous blood simulation box 4 and an arterial blood simulation box 5, a culture fluid injection port 6 is provided on the top of the multi-layer culture box 1, an arterial blood interface 7 and a venous blood interface 8 are provided on the upper side wall of the multi-layer culture box 1, a liquid outlet 9 is provided on the lower side wall of the multi-layer culture box 1, and the liquid outlet 9 is respectively connected to the venous blood simulation box 4 and the arterial blood simulation box 5 through a circulation tube 10, the microporous filter 2 and the circulating peristaltic pump 3 are sequentially arranged on the circulation tube 10, the venous blood simulation box 4 is connected to the venous blood interface 8 through a venous tube 51, the arterial blood simulation box 5 is connected to the arterial blood interface 7 through an arterial tube 52, a sample addition port 11 is provided on the venous blood simulation box 4, and the arterial blood simulation box 5 is also connected to an oxygen cylinder 12 and a carbon dioxide cylinder 13.

[0024] The multilayer incubator 1 comprises a box body 14 with an open front side, a box door 15 is hingedly connected to the open front side of the box body 14, a sealing strip 16 is arranged on the periphery of the inner side of the box door 15, and an arterial blood interface 7 and a venous blood interface 8 are arranged on the box door 15; a first heating element is arranged inside the inner wall of the box body 14, a first display screen 17 and a first microcontroller are arranged on the outer side of the box body 14, a first temperature sensor and a carbon dioxide concentration sensor are arranged on the inner wall of the box body 14, the first heating element, the first temperature sensor, the carbon dioxide concentration sensor and the first display screen 17 are respectively connected to the first microcontroller through lines, and the first heating element, the first temperature sensor, the carbon dioxide concentration sensor and the first microcontroller are all prior art and are not shown in the figure;

[0025] A rectangular support member and two groups of wedge-shaped support members are fixed in the box 14. The rectangular support member is located between the two groups of wedge-shaped support members. The rectangular support member is composed of two long prisms 18. The two prisms 18 are arranged side by side front and back. The two ends of the prisms 18 are respectively fixed on the left and right inner walls of the box 14. The wedge-shaped support member is composed of two wedge-shaped support edges 19. The two support edges 19 are arranged side by side front and back. The two ends of the support edges 19 are respectively fixed on the left and right inner walls of the box 14.

[0026] A top culture plate assembly 20 is placed on the uppermost wedge-shaped support, a middle culture plate assembly 21 is placed on the middle rectangular support, and a bottom culture plate assembly 22 is placed on the lowermost wedge-shaped support;

[0027] The top culture plate assembly 20 includes a first rectangular frame 23, the bottom surface of the first rectangular frame 23 is blocked by a bottom plate 24, the bottom plate 24 is an inclined structure and is lower on the left and higher on the right, the inclination of the bottom plate 24 is adapted to the inclination of the top surface of the supporting edge 19, a rectangular supporting inner frame 25 is fixed to the inner frame wall of the first rectangular frame 23, a rectangular pressing frame 26 is arranged in the first rectangular frame 23, the pressing frame 26 is located above the supporting inner frame 25, the front side of the pressing frame 26 is hinged to the front side of the first rectangular frame 23 through a pin 27, a torsion spring is arranged on the pin 27, a semipermeable membrane 28 is placed in the first rectangular frame 23, and the surroundings of the semipermeable membrane 28 are placed on the supporting inner frame 25 of the first rectangular frame 23, and under the action of the torsion spring, the pressing frame 26 is pressed on the surroundings of the semipermeable membrane 28;

[0028] The top culture plate assembly 20 has the same structure as the bottom culture plate assembly 22; two first perforations 29 are provided on the front side of the first rectangular frame 23 of the top culture plate assembly 20, and a venous blood inlet tube 30 and an arterial blood inlet tube 31 are placed on the upper surface of the semipermeable membrane 28. The venous blood inlet tube 30 and the arterial blood inlet tube 31 are placed side by side. One end of the venous blood inlet tube 30 passes through the first perforation 29 and is connected to the venous blood interface 8 of the box door 15, and one end of the arterial blood inlet tube 31 passes through another first perforation 29 and is connected to the arterial blood interface 7 of the box door 15. A plurality of first branches 32 are connected to the venous blood inlet tube 30 in parallel, and a plurality of second branches 33 are connected to the arterial blood inlet tube 31 in parallel. The plurality of first branches and second branches are arranged one by one, and a plurality of liquid outlet holes 34 are provided on the opposite surfaces of the first branch 32 and the adjacent second branch 33; a plurality of first diversion interfaces 35 are provided on the left side of the bottom plate 24;

[0029] A plurality of first through holes 36 are provided on the right side frame of the first rectangular frame 23 of the bottom culture plate assembly 22. The first through holes 36 are in an L-shaped structure. A second flow guide interface 37 is installed at the upper end of each first through hole 36. A plurality of third flow guide interfaces 38 are provided on the left side of the bottom plate 24 of the bottom culture plate assembly 22. The plurality of third flow guide interfaces 38 are connected to the liquid outlet 9 of the lower side wall of the multi-layer culture box 1 through pipelines.

[0030] The middle culture plate assembly 21 includes a second rectangular frame 39, and the front frame wall and the rear frame wall of the second rectangular frame 39 are both fixed with a long support plate 40, and the rear frame wall of the second rectangular frame 39 is provided with an arc groove 41, and the arc groove 41 is located above the long support plate 40, and the right frame wall of the second rectangular frame 39 is provided with a plurality of second through holes 42, and the left frame wall of the second rectangular frame 39 is provided with a plurality of third through holes, and the second through holes 42 and the third through holes are both L-shaped structures and are located above the long support plate 40, and the lower end of the second through hole 42 is installed with a fourth guide interface, and the upper end of the third through hole is installed with a fifth guide interface 43;

[0031] The several first flow guide interfaces 35 of the top culture plate assembly 20 are respectively connected to the several fifth flow guide interfaces 43 of the middle culture plate assembly 21 through the flow guide hose, and the fourth flow guide interface of the middle culture plate assembly 21 is respectively connected to the several second flow guide interfaces 37 of the bottom culture plate assembly 22 through the flow guide hose.

[0032] The venous blood simulation box 4 includes a box body 44, the inner cavity of the box body 44 is divided into a solution cavity 45 and a component installation cavity by a partition, a peristaltic pump 46 is installed in the component installation cavity, a second display screen 47 and a second microcontroller are arranged on the outer side of the box body 44, a pH sensor 48, a second temperature sensor 49 and a blood oxygen saturation probe 50 are arranged in the solution cavity 45, a second heating element is arranged on the inner wall at the bottom of the solution cavity 45 of the box body 44, the peristaltic pump 46, the second display screen 47, the pH sensor 48, the blood oxygen saturation probe 50, the second temperature sensor 49 and the second heating element are respectively connected to the second microcontroller through lines, and the liquid inlet pipe of the peristaltic pump 46 is connected to the bottom of the solution cavity 45; the sample addition port 11 is arranged at the top of the solution cavity 45 of the venous blood simulation box 4;

[0033] The structure of the arterial blood simulation box 5 is the same as that of the venous blood simulation box 4. The oxygen cylinder 12 and the carbon dioxide cylinder 13 are connected to the bottom of the solution chamber 45 of the arterial blood simulation box 5 through pipes. The circulation tube 10 is connected to the upper part of the solution chamber 45 of the arterial blood simulation box 5 and the venous blood simulation box 4 respectively. The outlet pipe of the peristaltic pump 46 of the arterial blood simulation box 5 is connected to the arterial tube 52, and the outlet pipe of the peristaltic pump 46 of the venous blood simulation box 4 is connected to the venous tube 51.

[0034] When the present invention performs liver cell mimetic culture, the culture fluid is firstly poured in from the culture fluid injection port 6 at the top of the multi-layer culture box 1, flows out after passing through the top culture plate assembly 20, the middle culture plate assembly 21 and the bottom culture plate assembly 22, and then enters the circulation tube 10. The microporous filter 2 filters out the dead cells in the culture fluid. Then, driven by the circulating peristaltic pump 3, the culture fluid enters the solution chamber 45 of the venous blood simulation box 4 and the arterial blood simulation box 5 respectively. New culture fluid, growth factors and serum can be added to the solution chamber 45 of the venous blood simulation box 4 through the sample addition port 11. The arterial blood simulation box 5 is connected to an oxygen cylinder 12 and a carbon dioxide cylinder 13 for replenishing the solution chamber 45 of the arterial blood simulation box 5. The amount of oxygen and carbon dioxide; the arterial blood simulation box 5 and the venous blood simulation box 4 are both provided with a second heating element, a pH sensor 48, and a blood oxygen saturation probe 50, the second heating element is used to heat the temperature in the solution cavity 45, the pH sensor 48 and the blood oxygen saturation probe 50 are respectively used to monitor the pH value and blood oxygen saturation of the culture solution in the solution cavity 45, and transmit the monitoring data to the second microcontroller, and the second microcontroller transmits the data to the second display screen 47 for display, under the action of the peristaltic pump 46, the culture solution in the solution cavity 45 of the venous blood simulation box 4 flows into the multi-layer incubator 1 through the venous tube 51, and the culture solution in the solution cavity 45 of the arterial blood simulation box 5 flows into the multi-layer incubator 1 through the arterial tube 52;

[0035] The multi-layer incubator 1 of the present invention is provided with a first display screen 17, a first heating element, a first temperature sensor and a carbon dioxide concentration sensor, which monitor the temperature and carbon dioxide concentration in the multi-layer incubator 1, and feed back the monitoring data to the first display screen 17 for display. The circulating culture fluid enters the arterial blood input tube 31 through the arterial blood interface 7, and another part of the culture fluid enters the venous blood input tube 30 through the venous blood interface 8. Finally, the culture fluid flows from the liquid outlet 34 to the semipermeable membrane 28 of the top culture plate assembly 20, on which the hepatic sinusoidal endothelial cells are planted. The culture fluid is filtered through the semipermeable membrane 28 and the hepatic sinusoidal endothelial cells and flows to the bottom plate 24. The bottom plate 24 is a slope structure. Under the guidance of the bottom plate 24, the liquid passes through the first guide The interface 35 and the fifth flow guide interface 43 flow to the middle culture plate assembly 21. A plurality of glass slides are placed on the two long support plates 40 of the middle culture plate assembly 21. The plurality of glass slides are arranged side by side, and the adjacent glass slides are close to each other. The arc-shaped groove 41 is convenient for placing or removing the glass slides. Liver cells are cultured on the glass slides. The liquid flows to the surface of the glass slide through the fifth flow guide interface 43, and then the liquid flows into the semipermeable membrane 28 of the bottom culture plate assembly 22 through the fourth flow guide interface and the second flow guide interface 37. Tumor cells are planted on the semipermeable membrane 28. After being filtered by the semipermeable membrane 28, the liquid flows out from the third flow guide interface 38 and finally flows out from the liquid outlet 9 of the multi-layer culture box 1. The outflowing liquid enters the microporous filter 2, so as to perform cyclic culture.

[0036] The first heating element and the second heating element in the present invention are electric heating wires, which belong to the existing conventional technology, and the specific structure and assembly and installation thereof are not described in detail.

[0037] This embodiment does not impose any formal limitation on the shape, material, structure, etc. of the present invention. Any simple modification, equivalent change and modification made to the above embodiment based on the technical essence of the present invention shall fall within the protection scope of the technical solution of the present invention.

Claims

1. A cell co-culture system simulating hepatic sinusoids, characterized in that: It comprises a multi-layer incubator, a microporous filter, a circulating peristaltic pump, a venous blood simulation box and an arterial blood simulation box, wherein a culture fluid injection port is provided on the top of the multi-layer incubator, an arterial blood interface and a venous blood interface are provided on the upper side wall of the multi-layer incubator, a liquid outlet is provided on the lower side wall of the multi-layer incubator, the liquid outlet is respectively connected to the venous blood simulation box and the arterial blood simulation box through a circulating tube, the microporous filter and the circulating peristaltic pump are sequentially arranged on the circulating tube, the venous blood simulation box is connected to the venous blood interface through a venous tube, the arterial blood simulation box is connected to the arterial blood interface through an arterial tube, a sample addition port is provided on the venous blood simulation box, and the arterial blood simulation box is also connected to an oxygen cylinder and a carbon dioxide cylinder; The multi-layer incubator comprises a box body with an open front side, a box door is hinged at the open front side of the box body, a sealing strip is arranged on the periphery of the inner side of the box door, and an arterial blood interface and a venous blood interface are arranged on the box door; a first heating element is arranged inside the inner wall of the box body, a first display screen and a first microcontroller are arranged on the outer side of the box body, a first temperature sensor and a carbon dioxide concentration sensor are arranged on the inner wall of the box body, and the first heating element, the first temperature sensor, the carbon dioxide concentration sensor and the first display screen are respectively connected to the first microcontroller through lines; A rectangular support member and two groups of wedge-shaped support members are fixed in the box, the rectangular support member is located between the two groups of wedge-shaped support members, the rectangular support member is composed of two long strip prisms, the two prisms are arranged side by side front and back, and the two ends of the prisms are respectively fixed on the left and right inner walls of the box, and the wedge-shaped support member is composed of two wedge-shaped support edges, the two support edges are arranged side by side front and back, and the two ends of the support edges are respectively fixed on the left and right inner walls of the box; The top layer culture plate assembly is placed on the uppermost wedge-shaped support, the middle layer culture plate assembly is placed on the middle rectangular support, and the bottom layer culture plate assembly is placed on the lowermost wedge-shaped support; The top culture plate assembly comprises a first rectangular frame, the bottom surface of the first rectangular frame is blocked by a bottom plate, the bottom plate is an inclined structure and is lower on the left and higher on the right, the inclination of the bottom plate is adapted to the inclination of the top surface of the supporting edge, a rectangular supporting inner frame is fixed to the inner frame wall of the first rectangular frame, a rectangular pressing frame body is arranged in the first rectangular frame, the pressing frame body is located above the supporting inner frame, the front side of the pressing frame body is hinged to the front side of the first rectangular frame through a pin shaft, a torsion spring is arranged on the pin shaft, a semipermeable membrane is arranged in the first rectangular frame, the periphery of the semipermeable membrane is arranged on the supporting inner frame of the first rectangular frame, and under the action of the torsion spring, the pressing frame body is pressed on the periphery of the semipermeable membrane; The top culture plate assembly has the same structure as the bottom culture plate assembly; two first perforations are provided on the front side of the first rectangular frame of the top culture plate assembly; a venous blood inlet tube and an arterial blood inlet tube are placed on the upper surface of the semipermeable membrane; the venous blood inlet tube and the arterial blood inlet tube are placed side by side on the left and right; one end of the venous blood inlet tube passes through the first perforation and is connected to the venous blood interface of the box door; one end of the arterial blood inlet tube passes through another first perforation and is connected to the arterial blood interface of the box door; a plurality of first branches are connected to the venous blood inlet tube in parallel; a plurality of second branches are connected to the arterial blood inlet tube in parallel; a plurality of first branches and second branches are arranged one by one, and a plurality of liquid outlet holes are provided on the opposite surfaces of the first branches and the adjacent second branches; a plurality of first diversion interfaces are provided on the left side of the bottom plate; A plurality of first through holes are provided on the right side frame of the first rectangular frame of the bottom culture plate assembly, the first through holes are in an L-shaped structure, a second flow guide interface is installed on the upper end of each first through hole, and a plurality of third flow guide interfaces are provided on the left side of the bottom plate of the bottom culture plate assembly, and the plurality of third flow guide interfaces are connected to the liquid outlet of the lower side wall of the multi-layer culture box through pipelines; The middle culture plate assembly includes a second rectangular frame, the front frame wall and the rear frame wall of the second rectangular frame are both fixed with a long support plate, the rear frame wall of the second rectangular frame is provided with an arc groove, the arc groove is located above the long support plate, the right frame wall of the second rectangular frame is provided with a plurality of second through holes, the left frame wall of the second rectangular frame is provided with a plurality of third through holes, the second through holes and the third through holes are both L-shaped and located above the long support plate, the lower end of the second through hole is provided with a fourth flow guide interface, and the upper end of the third through hole is provided with a fifth flow guide interface; The first flow guide interfaces of the top culture plate assembly are connected to the fifth flow guide interfaces of the middle culture plate assembly through the flow guide hoses, and the fourth flow guide interfaces of the middle culture plate assembly are connected to the second flow guide interfaces of the bottom culture plate assembly through the flow guide hoses; The venous blood simulation box comprises a box body, the inner cavity of the box body is divided into a solution cavity and a component installation cavity by a partition, a peristaltic pump is installed in the component installation cavity, a second display screen and a second microcontroller are arranged on the outer side of the box body, a pH sensor, a second temperature sensor and a blood oxygen saturation probe are arranged in the solution cavity, a second heating element is arranged on the inner wall at the bottom of the solution cavity of the box body, the peristaltic pump, the second display screen, the pH sensor, the blood oxygen saturation probe, the second temperature sensor and the second heating element are respectively connected to the second microcontroller through lines, and the liquid inlet pipe of the peristaltic pump is connected to the bottom of the solution cavity; a sample addition port is arranged at the top of the solution cavity of the venous blood simulation box; The structure of the arterial blood simulation box is the same as that of the venous blood simulation box. The oxygen cylinder and the carbon dioxide cylinder are connected to the bottom of the solution chamber of the arterial blood simulation box through pipes, the circulation tube is connected to the upper part of the solution chamber of the arterial blood simulation box and the venous blood simulation box respectively, the outlet tube of the peristaltic pump of the arterial blood simulation box is connected to the arterial tube, and the outlet tube of the peristaltic pump of the venous blood simulation box is connected to the venous tube.

Images