Organ culture system and device for preserving, repairing and regenerating in-vitro liver

Abstract

The invention relates to an organ culture system and device for preserving, repairing and regenerating an in-vitro liver. Comprising a pulse type centrifugal pump, a first constant flow centrifugal pump, a second constant flow centrifugal pump, an elastic liquid storage device, a bile collector, a first oxygenator, a second oxygenator, a blood storage device, a toxin molecule adsorption dialysis device, a blood gas analysis device, an intelligent organ preservation box, a display screen, a first leukocyte, thrombus and air filter and a second leukocyte, thrombus and air filter. A heparin pump pipe and a display; an antibiotic injection device, an insulin injection device, a glucagon and sodium bicarbonate injection device, a blood replacement injection inlet and a blood replacement outlet are arranged on the blood storage device; meanwhile, a main system and 13 subsystems are arranged, intelligent operation is achieved, the environment where the liver is located in vivo is bionic, long-time preservation and culture of isolated organs can be achieved, and the system is used for organ function evaluation, novel medicine exploration, edge organ repair, organ regeneration promotion and the like.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to an organ culture system and device for preservation, repair and regeneration of an ex vivo liver. Background Art

[0002] The shortage of donor livers is a major factor limiting the development of liver transplantation. According to data from the China Organ Donation Administration Center, approximately 300,000 people await organ transplants each year, but only a little over 10,000 are ultimately eligible. Many patients with end-stage liver disease die while waiting. Further expanding the supply of donor livers, repairing marginal organs, reducing surgical risks, and alleviating the organ shortage remain bottlenecks we must strive to overcome.

[0003] Normothermic machine perfusion (NMP), a new organ preservation technology that has emerged in recent years, has achieved numerous successes in the field of liver preservation and repair. Among the achievements to date, NMP primarily impacts liver donors in six key areas: 1. Assessing liver quality; 2. Prolonging liver preservation; 3. Reducing liver ischemia-reperfusion injury; 4. Repairing marginal donor livers; 5. Facilitating drug therapy and intervention; and 6. Improving post-transplant liver function. Based on this technology and system, NMP can provide a guarantee for expanding the liver donor pool.

[0004] With improvements in normothermic mechanical perfusion technology, the preservation time limit for isolated organs at room temperature has gradually been broken, and the longest preservation time record has been continuously refreshed. One research team performed normothermic mechanical perfusion after liver separation in vitro, with the longest half-liver perfusion time reaching 14 days, but the median liver viability was only 125 hours. These results demonstrate the limitations of commercially available normothermic mechanical perfusion systems. They are unable to provide the physiological conditions required for normal liver metabolism in vitro, and the liver perfusion time is limited, which cannot meet the needs of long-term repair and regeneration of isolated livers in vitro and "organ culture."

[0005] Currently, the main limitations on long-term ex vivo preservation of ex vivo livers include hemolysis, thrombosis, internal environmental disturbances, hepatocyte metabolic disorders, infection, and the accumulation of inflammatory factors and toxins. When the perfusion system environment does not match the physiological needs of the liver, hemodynamic instability can occur. After prolonged perfusion, the liver will exhibit symptoms such as swelling and increased resistance. Currently available normothermic liver mechanical perfusion technology lacks biomimetic capabilities and inherent system design flaws, making it unable to meet the needs of extremely long-term ex vivo preservation. Summary of the Invention

[0006] The object of the present invention is to provide an organ culture system and device for preservation, repair and regeneration of an isolated liver.

[0007] In order to achieve the above object, the present invention provides the following technical solutions: An organ culture system and device for the preservation, repair and regeneration of an ex vivo liver, comprising a pulse centrifugal pump, a first constant flow centrifugal pump, a second constant flow centrifugal pump, an elastic liquid reservoir, a bile collector, a first oxygenator, a second oxygenator, a blood reservoir, a toxin molecule adsorption dialysis device, a blood gas analysis device, an intelligent organ preservation box, a display screen, a first white blood cell, thrombus and air filter, a second white blood cell, thrombus and air filter, a heparin pump tube and a display.

[0008] The blood reservoir is provided with an antibiotic injection device, an insulin injection device, a glucagon injection device, a sodium bicarbonate injection device, a blood replacement injection inlet, and a blood replacement outflow port; the blood reservoir is provided with perfusion fluid (blood); the toxin molecule adsorption dialysis device and the blood gas analysis device are connected to the blood reservoir pipeline via a heparin pump tube; the intelligent organ preservation box contains a liver; one end of the hepatic artery catheter, the portal vein catheter, the vena cava catheter, and the bile duct catheter are connected to the liver pipeline, and the other end is connected to the blood reservoir pipeline via a heparin pump tube.

[0009] The bile collector is connected to the bile duct catheter pipeline via a heparin pump tube; the hepatic artery catheter is also connected to the flow controller F1, pressure sensor P1, oxygen saturation probe O1, pulse centrifugal pump, first white blood cell, thrombus and air filter and first oxygenator pipeline via the heparin pump tube; the portal vein catheter is also connected to the flow controller F2, pressure sensor P2, oxygen saturation probe O2, second white blood cell, thrombus and air filter and second oxygenator pipeline via the heparin pump tube; the vena cava catheter is also connected to the flow controller F3, pressure sensor P3, elastic liquid reservoir and second constant flow centrifugal pump pipeline via the heparin pump tube; the intelligent organ preservation box includes an intelligent organ preservation box shell, an intelligent organ preservation box top cover, an internal liquid storage device of the intelligent organ preservation box, an organ carrying net bag and a water bath outlet.

[0010] The intelligent organ preservation box is further provided with a peristaltic pump, a first perfusion line heating device, a water flow power outlet pipe, a water flow power return suction pipe, an intelligent organ preservation box water bath liquid inlet, an intelligent organ preservation box water bath liquid outlet, an internal circulation perfusion line, a second perfusion line heating device, and a temperature control probe on the internal liquid storage device of the intelligent organ preservation box. The intelligent organ preservation box is provided with preservation liquid. The intelligent organ preservation box balances the upward and downward forces exerted on the liver, thereby achieving suspended preservation of the organ (liver) and preservation at room temperature in vitro.

[0011] Preferably, the blood reservoir is an elastic sac device.

[0012] Preferably, the first leukocyte, thrombus and air filter and the second leukocyte, thrombus and air filter are hollow structures, and filter screens and leukocyte removers are provided in the first leukocyte, thrombus and air filter and the second leukocyte, thrombus and air filter.

[0013] Preferably, the portal vein catheter is further provided with an infrahepatic vena cava vascular clamp.

[0014] Preferably, the pressure range of the elastic liquid reservoir is 5-12 cmH2O.

[0015] Preferably, the pressure range of portal vein catheterization is 13-24 cmH2O, and the flow rate is set at 0.7-1.0 ml / g / min according to the liver weight.

[0016] Preferably, the pressure range of hepatic artery catheterization is 75-105 mmHg, the pulse frequency is 60-100 bpm, and the flow rate is set at 0.2-0.3 ml / g / min according to the liver weight.

[0017] Preferably, it includes a central control system, which includes 13 subsystem internal feedback loop control systems, and the 13 subsystem internal feedback loop control systems include a temperature control system, a pipeline pressure and flow sensing and regulation system, a blood glucose stabilization regulation system, a pH regulation system, a blood oxygen saturation control system, an electrolyte balance system, an inflammatory mediator monitoring and removal system, a toxin molecule dialysis adsorption system, an antibiotic injection system, a nutrient and trace element injection system, a liver surface pressure monitoring system, a coagulation function monitoring system and a thrombus removal system.

[0018] The beneficial effects of the present invention are: 1. The overall system adopts a bionic design to simulate the physiological environment of the liver in the body, including temperature, pressure, flow, oxygen saturation, cardiac cycle, and health of the perfusate (blood). This maintains system stability, reduces mechanical system-related liver damage, and provides a repair and regeneration support platform for long-term preservation of the liver in vitro and organ regeneration.

[0019] 2. For the first time, each module was systematized, 13 subsystems were integrated, and the intelligent operation of the overall system was realized, achieving automation, visualization, and scientific perfusion of organs.

[0020] 3. The intelligent organ preservation box adopts a suspension state similar to that of a baby in amniotic fluid, innovatively using the upward power of the water bath to offset the effect of gravity, thereby minimizing the compressive damage caused by the liver's own gravity.

[0021] 4. Constant flow centrifugal pumps are used in the portal vein and superior and inferior vena cava, and pulse centrifugal pumps are used in the hepatic artery to simulate the hemodynamic characteristics in vivo.

[0022] 5. A unique liver internal pressure regulation device coordinates the regulation of liver internal pressure through a second constant-flow centrifugal pump and elastic water bladder in the superior and inferior vena cava, a pulse centrifugal pump in the hepatic artery, and a first constant-flow centrifugal pump in the portal vein. This creates a mutual feedback loop to control perfusion pressure and flow, ensuring adequate and uniform perfusion of the liver. This ensures a more physiological state, reproducing the pressure and flow changes that the liver experiences during the cardiac cycle in vivo.

[0023] The above description is only an overview of the technical solution of the present invention. In order to more clearly understand the technical means of the present invention and implement it according to the contents of the specification, the following is a detailed description of the preferred embodiments of the present invention with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a schematic diagram of the main structure of the present invention; Figure 2 Distribution of subsystems of the present invention; Figure 3 It is a schematic diagram of the side view structure of the present invention; Figure 4 It is a schematic diagram of the top view structure of the present invention; Figure 5 This is a schematic diagram of the liver force shown in the present invention.

[0025] In the figure: 1-pulse centrifugal pump; 2-first constant flow centrifugal pump; 3-second constant flow centrifugal pump; 4-elastic liquid reservoir; 5-bile collector; 6-first oxygenator; 7-second oxygenator; 8-blood reservoir; 9-toxin molecule adsorption dialysis device; 10-blood gas analysis device; 11-intelligent organ preservation box; 12-display screen; 13-first white blood cell, thrombus and air filter; 14-second white blood cell, thrombus and air filter; 15-antibiotic injection device; 16-insulin injection device; 17-glucagon; 18-sodium bicarbonate injection device; 19-hepatic artery catheter; 20-portal vein catheter; 21-vena cava catheter; 22-biliary tract catheter Tube; 23-liver; 24-heparin-coated pump tube; 25-blood exchange outflow port; 26-blood exchange injection inlet; 1101-smart organ preservation box housing; 1102-peristaltic pump; 1103-first perfusion line heating device; 1104-water flow power outlet pipe; 1105-water flow power return suction pipe; 1106-water bath liquid injection port; 1107-water bath liquid discharge port; 1108-organ carrying net bag; 1109-water bath outlet hole; 1110-internal liquid storage device; 1111-internal circulation perfusion line; 1112-second perfusion line heating device; 1113-smart organ preservation box top cover; 1114-temperature control probe. DETAILED DESCRIPTION

[0026] The technical solution of the present invention will be clearly and completely described below in conjunction with the accompanying drawings. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. In the description of the present invention, it should be noted that the orientation or position relationship indicated by the terms "center", "up", "down", "left", "right", "vertical", "horizontal", "inside", "outside", etc. is based on the orientation or position relationship shown in the accompanying drawings, which is only for the convenience of describing the present invention and simplifying the description, 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. In addition, the terms "first", "second", and "third" are only used for descriptive purposes and cannot be understood as indicating or implying relative importance.

[0027] In the description of the present invention, it should be noted that, unless otherwise expressly specified or limited, the terms "mounted," "connected," and "connected" should be understood in a broad sense. For example, they may refer to fixed, detachable, or integral connections; mechanical or electrical connections; direct or indirect connections through an intermediate medium; and internal communication between two components. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on the specific circumstances.

[0028] In addition, the technical features involved in the different embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0029] like Figure 1-Figure 5 As shown, an organ culture system and device for preservation, repair and regeneration of an ex vivo liver comprises a pulse centrifugal pump 1, a first constant flow centrifugal pump 2, a second constant flow centrifugal pump 3, an elastic liquid reservoir 4, a bile collector 5, a first oxygenator 6, a second oxygenator 7, a blood reservoir 8, a toxin molecule adsorption dialysis device 9, a blood gas analyzer 10, an intelligent organ preservation box 11, a display screen 12, a first white blood cell, thrombus and air filter 13, a second white blood cell, thrombus and air filter 14, a heparin pump tube and a display; the blood reservoir 8 is provided with an antibiotic injection device 15, an insulin injection device 16, a glucagon injection device 17, a sodium bicarbonate injection device 18, a blood replacement injection inlet 26, and a blood replacement outflow port 25; the blood reservoir 8 is provided with a perfusate (blood); the toxin molecule adsorption dialysis device 9 and the blood gas analyzer 10 are connected to the blood reservoir 8 via a heparin pump tube; The intelligent organ preservation box 11 contains a liver 23; one end of the hepatic artery catheter 19, the portal vein catheter 20, the vena cava catheter 21, and the bile duct catheter 22 are connected to the liver 23 pipeline, and the other end is connected to the blood reservoir 8 pipeline through a heparin pump tube; the hepatic artery catheter 19 is connected to the hepatic artery of the liver 23, the portal vein catheter 20 is connected to the portal vein of the liver 23, and the vena cava catheter 21 is connected to the vena cava of the liver 23; the bile collector 5 is connected to the bile duct catheter 22 pipeline through a heparin pump tube, and the bile collector 5 is used to collect bile produced by the liver 23; the hepatic artery catheter 19 is connected to the hepatic artery of the liver 23, the portal vein catheter 20 is connected to the portal vein of the liver 23, and the vena cava catheter 21 is connected to the vena cava of the liver 23. The catheter 20 is also connected to the flow controller F1, pressure sensor P1, oxygen saturation probe O1, pulse centrifugal pump 1, first white blood cell, thrombus and air filter 13 and first oxygenator 6 pipelines through a heparin pump tube; the portal vein catheter 20 is also connected to the flow controller F2, pressure sensor P2, oxygen saturation probe O2, second white blood cell, thrombus and air filter 14 and second oxygenator 7 pipelines through a heparin pump tube; the vena cava catheter 21 is also connected to the flow controller F3, pressure sensor P3, elastic liquid reservoir 4 and second constant flow centrifugal pump 3 pipelines through a heparin pump tube.

[0030] Display screen 12 serves as an observation window for the organ culture system's operation. It serves as one of the control terminals for the central control system and its 13 subsystems. It can set and display various physiological and perfusion parameters of the liver 23 in real time, useful for setting up the culture system and monitoring organ function. Simultaneously, data is transmitted via the internet to a mobile phone terminal, displaying various organ parameters in real time to ensure stable operation.

[0031] The hepatic artery catheter 19, controller F1, pressure sensor P1, oxygen saturation probe O1, pulse centrifugal pump 1, first white blood cell, thrombus and air filter 13 and first oxygenator 6 constitute a first perfusion circulation system; the portal vein catheter 20, flow controller F2, pressure sensor P2, oxygen saturation probe O2, second white blood cell, thrombus and air filter 14 and second oxygenator 7 constitute a second perfusion circulation system; the vena cava catheter 21, flow controller F3, pressure sensor P3, elastic fluid reservoir 4 and second constant flow centrifugal pump 3 constitute a third perfusion circulation system; the first perfusion circulation system, the second perfusion circulation system and the third perfusion circulation system together constitute an intrahepatic pressure and flow regulation device, and together complete the bionic perfusion of the liver 23. The first perfusion circulation system and the second perfusion circulation system are the blood inflow channels of the liver 23. The perfusion fluid (blood) enters the liver 23 through the first perfusion circulation system and the second perfusion circulation system. The third perfusion circulation system is the blood outflow channel of the liver 23. Through the cooperation of the first perfusion circulation system, the second perfusion circulation system and the third perfusion circulation system, the blood flow and internal pressure inside the liver 23 can be jointly regulated, and the flow meter pressure of the liver 23 perfusion can be finely regulated, especially for the uniform perfusion of the liver 23. An elastic fluid reservoir 4 and a constant flow centrifugal pump are added to the outflow channel of the liver 23 to simulate physiological conditions. During the heartbeat, the pressure of the superior and inferior vena cava of the liver fluctuates to realize the regulation of the outflow channel resistance of the liver 23. The first perfusion circulation system and the second perfusion circulation system are combined to jointly regulate the pressure inside the liver 23, thereby making the perfusion of the liver 23 more sufficient.

[0032] The smart organ preservation box 11 includes a smart organ preservation box housing 1101, a smart organ preservation box top cover 1113, and a liquid storage device 1110, an organ carrying net bag 1108, and a water bath outlet 1109 within the smart organ preservation box 11. The liquid storage device 1110 within the smart organ preservation box 11 is also equipped with a peristaltic pump 1102, a first perfusion pipeline heating device 1103, a water flow power outlet pipe 1104, a water flow power return suction pipe 1105, a water bath liquid inlet 1106 for the smart organ preservation box 11, a water bath liquid outlet 1107 for the smart organ preservation box 11, an internal circulation perfusion pipeline 1111, a second perfusion pipeline heating device 1112, and a temperature control probe 1114. The smart organ preservation box 11 is provided with a preservation fluid, which is a water bath fluid (peritoneal dialysis fluid).

[0033] Preferably, the blood reservoir 8 is an elastic capsule device that simulates the elastic blood reservoir function of the large blood vessels and venous network in the body.

[0034] Preferably, the first leukocyte, thrombus and air filter 13 and the second leukocyte, thrombus and air filter 14 are hollow structures, and filter screens and leukocyte removers are provided in the first leukocyte, thrombus and air filter 13 and the second leukocyte, thrombus and air filter 14 for removing bubbles and thrombi.

[0035] Preferably, the portal vein catheter 20 is further provided with an infrahepatic vena cava clamp, which is used to clamp the infrahepatic vena cava of the liver 23 after the liver 23 is placed in the intelligent organ preservation box 11 .

[0036] Preferably, the pressure range of the elastic reservoir 4 is 5-12 cmH2O. At the beginning of perfusion, the pressure of the elastic reservoir 4 is 0. As the perfusion fluid (blood) flows out of the liver 23, the pressure of the elastic reservoir 4 gradually increases. When the pressure of the elastic reservoir 4 rises to 12 cmH2O, the flow rate of the constant flow centrifugal pump increases, accelerating the outflow rate of the perfusion fluid (blood).

[0037] Preferably, the pressure range of the portal vein catheter 20 is 13-24 cmH2O, and is set to 0.7-1.0 ml / g / min according to the weight of the liver (understood as 0.7-1.0 ml of blood per minute per gram of liver). Taking a liver with a mass of 1500 g as an example, the portal vein flow range is set to 1050-1500 ml / min.

[0038] Preferably, the pressure range of the hepatic artery catheter 19 is 75-105 mmHg, the pulse frequency is 60-100 bpm, and it is set to 0.7-1.0 ml / g / min according to the weight of the liver (understood as 0.7-1.0 ml of blood per minute per gram of liver). Taking a liver with a mass of 1500 g as an example, the hepatic artery pulse flow range is set to 300-450 ml / min.

[0039] Preferably, it also includes a central control system, which includes the establishment of internal feedback loop controls for 13 subsystems. The 13 subsystems include a temperature control system, a pipeline pressure and flow sensing and regulation system, a blood glucose stabilization and regulation system, a pH regulation system, a blood oxygen saturation control system, an electrolyte balance system, an inflammatory mediator monitoring and removal system, a toxin molecule dialysis adsorption system, an antibiotic injection system, a nutrient and trace element injection system, a liver surface pressure monitoring system, a coagulation function monitoring system, and a thrombus removal system. Each individual subsystem involves the coordination, monitoring, and intervention of the system including hardware and software, and each individual subsystem simultaneously realizes the automation and intelligent feedback control and operation of each subsystem.

[0040] During use: The intelligent organ preservation box and its tubing are pre-filled with preservation fluid (peritoneal dialysis fluid). After pre-filling the blood reservoir and tubing of the organ culture system with 37°C perfusate (blood), the organ's tubing is connected to the corresponding tubing of the culture system. In the main control system on the display screen, the initial flow rate of the hepatic artery and portal vein is set at 1 ml / g / min based on the organ weight, with a portal vein flow rate: hepatic artery flow rate ratio of 3:1. The organ culture system's tubing pressure and flow sensing and regulation system is activated. Driven by the pulsed centrifugal pump 1, the perfusate (blood) in the blood reservoir 8 enters the perfusion system through the heparin-coated pump tubing 24. It is oxygenated by the first oxygenator 6. The perfusate is then cleared of white blood cells, thrombi, and bubbles by the first white blood cell, thrombus, and air filter 13. The perfusate then enters the liver 23 through the pulsed centrifugal pump 1. Driven by a constant-flow centrifugal pump, the perfusate (blood) in blood reservoir 8 enters the perfusion system through heparin-coated pump tubing 24, passes through the second oxygenator 7 for oxygenation, and then passes through the second leukocyte, thrombus, and air filter 14 to remove white blood cells, thrombi, and bubbles from the perfusate. The blood then passes through the constant-flow centrifugal pump and enters liver 23. Blood in liver 23 enters blood reservoir 8 through the vena cava catheter 21, elastic reservoir 4, and constant-flow centrifugal pump. The first perfusion circulation system uses a pulsed centrifugal pump 1 to simulate cardiac pulsatile output. The second perfusion circulation system uses a constant-flow centrifugal pump to simulate the steady state of blood flow after the confluence of the mesenteric and splenic veins. The third perfusion circulation system simulates the pressure and flow changes in the vena cava in vivo, with a biomimetic design based on the pressure and flow changes of the vena cava as the heart contracts and relaxes.

[0041] At the same time, if Figure 3 and Figure 4 As shown, the liver surface pressure monitoring system in the organ culture system is activated. Sterile, room-temperature (36-37°C) organ bath fluid (peritoneal dialysis fluid) is injected into the intelligent organ preservation chamber 11 through the bath fluid inlet 1106 into the internal fluid storage device 1110. Waste organ bath fluid (peritoneal dialysis fluid) is discharged through the bath fluid outlet 1107. After the organ bath fluid is injected, the intelligent organ preservation chamber 11 is operated. The peristaltic pump 1102 drives the organ bath fluid (peritoneal dialysis fluid) through the system, passing through the hydraulic return suction pipe 1105, the peristaltic pump 1102, the internal circulation perfusion line 1111, and the hydraulic outlet pipe 1104 before finally flowing out of the bath outlet 1109 at the bottom of the intelligent organ preservation chamber 11, creating a vertical upward water flow impact force.

[0042] like Figure 5As shown, the thick arrow indicates the direction of water flow. The organ carrying net bag 1108 in the figure plays the role of carrying the liver 23 before the water bath power device is activated. The smart organ preservation box can meet the requirements of non-contact preservation of the liver, so that the liver is suspended in the preservation liquid, avoiding the compression damage caused by the interaction between the liver's own gravity and the carrier. When the liver 23 is placed in the smart organ preservation box 11, the external forces acting on the liver 23 are analyzed, which are the downward gravity G and the upward buoyancy F. 浮 , Net bag support force F 支 and water flow impact force F 冲 ; ; (See Figure 5 Therefore, the characteristic is that the gravity on the liver 23 is balanced with the upward water flow impact force, liquid buoyancy, and the net bag support force, so that the net bag's support force F on the liver 23 is 支 Infinitely approaching 0, thereby minimizing gravitational compression injuries.

[0043] The pressure on the liver 23 in the intelligent organ storage box 11 includes four types: the liver 23's own weight G, the water flow impact force F, 流 Liver 23 buoyancy F 浮 , Net bag support force F 支 . After the water bath power device is started and operated, driven by the peristaltic pump 1102, the vertical upward water flow impact force formed by the water bath outlet 1109 gradually increases. Combined with the buoyancy of the liver 23 in the organ water bath fluid (peritoneal dialysis fluid), the carrying capacity of the net bag on the liver 23 gradually decreases. When the water flow impact force F flow + buoyancy F float = liver 23 gravity mg, the liver 23 can be suspended in the organ water bath fluid (peritoneal dialysis fluid), and the supporting force of the net bag on the liver 23 is 0. The intelligent organ preservation box will protect the liver from gravity compression damage caused by traditional preservation technology, thereby achieving maximum protection for the organ.

[0044] For organ preservation, evaluation, repair, and regeneration, the basic process is the same: the organ culture system's tubing is prefilled with 37°C perfusion fluid (blood), the harvested liver is placed in the intelligent organ preservation chamber, and the corresponding tubing is connected and activated. For livers that require long-distance transportation, this organ culture system monitors and adjusts various parameters in real time to enable long-term ex vivo preservation. For livers with impaired organ function and the potential for primary liver dysfunction or early dysfunction, accurate and comprehensive assessment of organ function is achieved through analysis of various parameters. For marginal livers such as livers from cardiac death donors, fatty livers, elderly donors, and donors with benign tumors, damaged organs can be repaired through drug therapy, gene therapy, or surgical intervention administered through the perfusion system. For split or living donor liver transplantation, the split liver is placed in the system. The system operates over a long period of time, providing sufficient nutrients, oxygen, and blood to promote liver cell division, enabling partial liver regeneration and achieving multiple uses. The implementation of the above functions relies on the patented ultimate biomimetic design in perfusion technology and organ preservation, which can achieve ultra-long-term in vitro preservation of the liver, providing time guarantee for the preservation, repair and regeneration of the liver.

[0045] When the pressure of the elastic reservoir 4 is lower than 5cmH2O, the internal pressure of the liver 23 decreases, the overall resistance of the liver 23 decreases, the pressure of the hepatic artery and portal vein decreases, the flow rate increases, and the liver 23 is overperfused, with the risk of shear damage caused by high flow. At this time, the first constant flow centrifugal pump 2 and the second constant flow centrifugal pump 3 flow rate decreases or even stops to adjust the pressure and flow of the elastic reservoir 4, thereby adjusting the pressure and flow of the hepatic artery and portal vein to restore their physiological state. On the contrary, when the pressure of the elastic reservoir 4 is higher than 12cmH2O, the internal pressure of the liver 23 rises, the overall resistance of the liver 23 rises, the pressure of the hepatic artery and portal vein increases, the flow rate decreases, the liver 23 is underperfused, and thrombosis and microcirculatory disorders caused by low flow are formed. At this time, the first constant flow centrifugal pump 2 and the second constant flow centrifugal pump 3 flow rate increases to adjust the pressure and flow of the elastic reservoir 4, thereby adjusting the pressure and flow of the hepatic artery and portal vein to restore their physiological state.

[0046] When the sum of the flow of the first constant flow centrifugal pump 2 and the flow of the pulse centrifugal pump 1 is higher than the flow of the second constant flow centrifugal pump 3, the elastic reservoir 4 will expand, and the amount of fluid accumulation will increase. At the same time, the overall pressure of the liver 23 outflow tract will rise, so that the perfusion fluid entering the liver 23 through the portal vein perfusion tube will not flow out of the liver 23 too quickly, so that a certain amount of perfusion fluid will accumulate in the liver 23. When the pressure in the liver 23 reaches 12cmH2O, the speed of the second centrifugal pump will increase, and the outflow tract flow will increase. During the operation of the whole system, they will adjust to each other, thereby controlling the hepatic artery pressure. The pressure is controlled between 75 mmHg and 105 mmHg, with a pulse frequency between 60 and 100 bpm and a flow rate no higher than 400 ml / min. The portal vein pressure is controlled between 13 cmH2O and 24 cmH2O, with a flow rate no higher than 1600 ml / min. The pressure in the superior and inferior vena cava is controlled between 5 mmHg and 12 mmHg, with a flow rate no higher than 2000 ml / min. This cycle is repeated to achieve optimal perfusion of the entire liver. Similarly, the hepatic artery flow rate is set between 15 and 30 cm / s, and the portal vein flow rate is set between 10 and 20 cm / s. The feedback control principle is the same as above.

[0047] According to Bernoulli's equation, . The faster the flow rate, the lower the pressure. The slower the flow rate, the greater the pressure. In order to simulate the blood flow in the liver 23 blood vessels under physiological conditions as much as possible, the flow rate needs to be reduced as much as possible while ensuring the flow rate, which is beneficial for protecting the vascular endothelial cells. The pressure and flow settings of the portal vein and hepatic artery depend on the pressure and flow of the superior and inferior vena cava (catheterization site). If the pressure in the superior and inferior vena cava is reduced, the flow rate there will increase, accompanied by a decrease in pressure and an increase in flow rate in the portal vein and hepatic artery. The vascular shear force associated with high flow rate will cause additional damage. Therefore, the pressure in the superior and inferior vena cava should be as close as possible to the vena cava pressure of 5-12cmH2O, so that the pressure, flow, and flow rate of the portal vein, hepatic artery, and superior and inferior vena cava are as close as possible to the in vivo environment.

[0048] In summary, the present invention has biomimized the environment in which the liver resides in the body, providing a scientific platform for the preservation, evaluation, repair, and regeneration of organs in vitro. Specifically, it can: 1. Prolong the preservation time of donated human livers in vitro, achieve long-term preservation and transportation, buy time for donated organs and transplant recipients, and reduce the organ abandonment rate; 2. Monitor various liver parameters in real time through changes in synthesis, metabolism and other indicators, and accurately evaluate liver function; 3. Provide channels for repair methods such as drug therapy, gene therapy, and surgical treatment of damaged livers; 4. Change liver transplantation from emergency surgery to elective surgery, making the organ, patient, and surgical preparation more adequate and increasing surgical safety; 5. The biomimetic organ culture system can provide sufficient time for the regeneration of the split part of the liver, realize multiple uses of one liver, and ultimately expand the source of organs and save the lives of more patients with terminal diseases.

[0049] The technical features of the above-mentioned embodiments can be combined arbitrarily. In order to make the description concise, not all possible combinations of the technical features in the above-mentioned embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0050] The above-described embodiments merely represent several implementation methods of the present invention. While the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent. It should be noted that a person skilled in the art would be able to make various modifications and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be determined by the appended claims.

Claims

1. An organ culture system and device for preservation, repair and regeneration of an ex vivo liver, characterized in that: It includes a pulse centrifugal pump, a first constant flow centrifugal pump, a second constant flow centrifugal pump, an elastic liquid reservoir, a bile collector, a first oxygenator, a second oxygenator, a blood reservoir, a toxin molecule adsorption dialysis device, a blood gas analysis device, an intelligent organ preservation box, a display screen, a first white blood cell, a thrombus and air filter, a second white blood cell, a thrombus and air filter, a heparin pump tube and a display screen; The blood reservoir is equipped with an antibiotic injection device, an insulin injection device, a glucagon injection device, a sodium bicarbonate injection device, a blood replacement injection inlet, and a blood replacement outflow port; the blood reservoir is provided with perfusion fluid (blood); the toxin molecule adsorption dialysis device and the blood gas analysis device are connected to the blood reservoir pipeline via a heparin pump tube; the intelligent organ preservation box contains a liver; the hepatic artery catheter, portal vein catheter, vena cava catheter, and bile duct catheter are connected to the liver pipeline at one end and to the blood reservoir pipeline at the other end via a heparin pump tube; The bile collector is connected to the bile duct catheter pipeline through a heparin pump tube; the hepatic artery catheter is also connected to the flow controller F1, the pressure sensor P1, the oxygen saturation probe O1, the pulse centrifugal pump, the first white blood cell, the thrombus and the air filter and the first oxygenator pipeline through the heparin pump tube; the portal vein catheter is also connected to the flow controller F2, the pressure sensor P2, the oxygen saturation probe O2, the second white blood cell, the thrombus and the air filter and the second oxygenator pipeline through the heparin pump tube; the vena cava catheter is also connected to the flow controller F3, the pressure sensor P3, the elastic liquid reservoir and the second constant flow centrifugal pump pipeline through the heparin pump tube; The intelligent organ preservation box includes an intelligent organ preservation box shell, an intelligent organ preservation box top cover, an internal liquid storage device of the intelligent organ preservation box, an organ carrying net bag and a water bath outlet. The internal liquid storage device of the intelligent organ preservation box is also provided with a peristaltic pump, a first perfusion pipeline heating device, a water flow power outlet pipe, a water flow power return suction pipe, an intelligent organ preservation box water bath liquid injection port, an intelligent organ preservation box water bath liquid discharge port, an internal circulation perfusion pipeline, a second perfusion pipeline heating device and a temperature control probe; the intelligent organ preservation box is provided with preservation liquid.

2. The organ culture system and device for preservation, repair and regeneration of an ex vivo liver according to claim 1, characterized in that: The blood reservoir is an elastic sac device.

3. The organ culture system and device for preservation, repair and regeneration of an ex vivo liver according to claim 1, characterized in that: The first leukocyte, thrombus and air filter and the second leukocyte, thrombus and air filter are hollow structures, and filter screens and leukocyte removers are provided in the first leukocyte, thrombus and air filter and the second leukocyte, thrombus and air filter.

4. The organ culture system and device for preservation, repair and regeneration of an ex vivo liver according to claim 1, characterized in that: The portal vein catheter is also provided with an infrahepatic vena cava vascular clamp.

5. The bionic liver repair device based on normothermic mechanical perfusion technology according to claim 1, characterized in that: The pressure range of the elastic reservoir is 5-12cmH2O.

6. The organ culture system and device for preservation, repair and regeneration of an ex vivo liver according to claim 1, characterized in that: The pressure range of portal vein catheterization is 13-24 cmH2O, and the flow rate is set at 0.7-1.0 ml / g / min according to the liver weight.

7. The organ culture system and device for preservation, repair and regeneration of an ex vivo liver according to claim 1, characterized in that: The pressure range of hepatic artery catheterization is 75-105 mmHg, the pulse frequency is 60-100 bpm, and the flow rate is set at 0.2-0.3 ml / g / min according to the liver weight.

8. The organ culture system and device for preservation, repair and regeneration of an ex vivo liver according to claim 1, characterized in that: It includes a central control system, which is established by internal feedback loop control of 13 subsystems. The 13 subsystems include a temperature control system, a pipeline pressure and flow sensing and regulation system, a blood sugar stabilization and regulation system, a pH regulation system, a blood oxygen saturation control system, an electrolyte balance system, an inflammatory mediator monitoring and removal system, a toxin molecule dialysis adsorption system, an antibiotic injection system, a nutrient and trace element injection system, a liver surface pressure monitoring system, a coagulation function monitoring system and a thrombus removal system.

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