A system for storing or culturing organ or tissue models and uses thereof

Abstract

The invention relates to a system for storing or cultivating organ or tissue models and its use. The system is designed for verifying the functionality of the harvested organ or tissue models and allowing them to be processed. The system can be used for cultivating organ and / or tissue models harvested from donors and / or prepared using other techniques, such as using 3D and / or 4D bioprinting or electrospinning techniques, and comprises a chamber (2) for the organ or tissue models, which is equipped with means for measuring and adjusting the temperature of the perfusion liquid in the chamber, a perfusion liquid container (1), which is equipped with means for measuring and adjusting the temperature of the perfusion liquid therein, means (4) for measuring the glucose concentration in the perfusion liquid and adding glucose to the perfusion liquid in a metered amount, means (3) for measuring the pH of the perfusion liquid and adding a substance for adjusting the pH of the perfusion liquid in a metered amount, means for non-contact mixing of the perfusion liquid, and means for measuring and controlling the flow parameters of the perfusion liquid.

Description

Technical Field

[0001] This invention relates to a system for storing or culturing organ or tissue models having a flow system or vascular system, and its uses. The system is designed to, but is not limited to, verify the function of a harvested organ or tissue model and allow for its processing. The system can be used to culture organ or tissue models harvested from a donor and / or prepared using other techniques, such as 3D and / or 4D bioprinting and / or electrospinning. The system can be used for the flow culture of organs that cannot be harvested in a patient or otherwise processed. The term "organ flow system" refers to an artificially prepared organ, indicating a system having formed channels. The term "organ vascular system" refers to a vascular system with endothelial cells, either harvested from a donor or artificially prepared. The term "storage" refers to maintaining an organ or tissue model at a low temperature during reduced metabolism or before cell colonization. The term "culture" refers to maintaining an organ or tissue model colonized with cells at a normal body temperature environment corresponding to in vivo conditions. The term "tissue model" refers to a three-dimensional structure containing living cells suspended in biological material, which may additionally include a vascular system. Background Technology

[0002] US Patent No. 9756851B2 discloses a composition, method, and apparatus for maintaining the viability of a harvested organ prior to implantation. The organ perfusion apparatus includes a storage chamber for storing the organ. The perfusion circuit is provided with a first line for supplying oxygenated fluid to the organ and a second line for draining the used fluid from the organ. The perfusion apparatus also includes means functionally connected to a perfusion circuit for maintaining the organ at normal body temperature. Furthermore, the apparatus includes means for controlling the pressure of the perfusion fluid, an oxygenation device for oxygenating at least some of the fluid, a filtration device, and a flow control device for controlling the flow of at least some of the fluid.

[0003] Patent document WO1996029865A1 describes an organ perfusion device capable of perfusing an organ at near-normal temperatures using blood or other oxygen-carrying substances. This device allows for the assessment of organ activity using online measurements of physiological efficiency. Embodiments use a computer-controlled blood pump to characterize the physiological conditions of perfusion. The device includes components that allow for the replenishment of lost circulating volume and the infusion of nutrients, drugs, and perfusion fluid components to aid in maintaining or regenerating the organ. The device can also regulate pressure, pH, and temperature, automatically measure the production rates of urine, bile, pancreatic duct secretions, or other physiological secretions, and determine blood or perfusion fluid flow rates, vascular resistance, and organ swelling.

[0004] US2017339945A1 discloses an apparatus for multi-organ perfusion, the organs being selected from the group consisting of the heart, liver, kidney, and lung. The apparatus includes a basic unit configured to be detachably connected to a perfusion module for organ perfusion. The basic unit includes: tubing for connecting a perfusion fluid source to the organ, such that the perfusion fluid circulates through the organ; a first pump and a second pump connected to the tubing to drive the perfusion fluid to circulate within the tubing; and a controller configured and connected to control the first and second pumps to control the circulation of the perfusion fluid through the organ. The controller can control the first and second pumps based on perfusion parameters selected based on organ type to perform organ perfusion.

[0005] Document EP1879997A2 discloses a portable organ perfusion device having an organ chamber for supporting organ immersion in perfusion fluid. A pump circulates the perfusion fluid around a circuit containing the pump, a heat exchanger for cooling the perfusion fluid, an oxygenator for oxygenating the perfusion fluid, and a device for ensuring a constant supply of fluid to the organ's vascular system. A bypass channel provides a fluid connection, allowing excess fluid to bypass the organ. The device also includes an oxygen source, sensors, a power supply, and a control unit for receiving information from the sensors and providing control commands to a control device.

[0006] Solutions are being sought to ensure effective control over maintaining normal body temperature conditions, regulating fluids, or controlling pH to efficiently, more automatically, and accurately repair or culture organ and tissue models. The object of this invention is to provide suitable conditions for culturing and storing organs harvested from deceased donors, such as pancreas, liver, kidney, lung, heart, small intestine, large intestine, thyroid gland, skin, brain, or prepared using other techniques, such as 3D and / or 4D bioprinting, like printing tissue models with vascular systems. The term "bionic organ" refers to tissue models printed directly with cells using 3D and / or 4D techniques, as well as separate scaffolds (vascular systems and tissue models), which are then colonized with their respective cells. The term "3D bioprinting" refers to the use of additive manufacturing techniques (layer-by-layer material application) to form three-dimensional structures containing living cells. In 4D bioprinting, the fourth dimension is time; the technique produces strips of material that are later transformed into a previously defined object (shape). Summary of the Invention

[0007] In a first aspect of the invention, a system is provided for storing or culturing organ or tissue models, said organ or tissue models preferably collected from a donor, preferably selected from pancreas, liver, kidney, lung, heart, small intestine, large intestine, thyroid gland, skin, brain, or prepared using other techniques, preferably using 3D bioprinting and / or 4D bioprinting and / or electrospinning techniques, preferably in the form of organ and / or tissue models printed with a flow system or vascular system, said system comprising:

[0008] - A chamber for an organ or tissue model, said chamber being equipped with means for measuring and regulating the temperature of the perfusion fluid placed in said chamber during operation of said system, wherein said means for measuring and regulating the temperature of said perfusion fluid preferably includes a water jacket.

[0009] - An infusion fluid container equipped with means for measuring and regulating the temperature of the infusion fluid in the container during system operation, preferably including a heating-cooling plate, wherein the infusion fluid container is connected to the organ chamber via at least one first line and at least one second line, during operation the infusion fluid can flow from the infusion fluid container to the organ chamber through the first line, and during operation the infusion fluid can flow from the organ chamber to the infusion fluid container through the second line.

[0010] - A device for measuring the glucose concentration in the perfusion solution and for adding glucose to the perfusion solution in a metered manner.

[0011] - A device for measuring the pH of the infusion solution and for adding, in metered amounts, a substance for adjusting the pH of the infusion solution.

[0012] - A device for non-contact mixing of the infusion fluid,

[0013] - A device for measuring and controlling the flow parameters of the infusion fluid, said flow parameters including infusion fluid pressure and / or infusion fluid output expressed as a unit volume per unit time.

[0014] - Optionally, an apparatus for measuring and controlling one or more parameters selected from: lactate concentration in the perfusion fluid, sodium ion concentration in the perfusion fluid, chloride ion concentration in the perfusion fluid, potassium ion concentration in the perfusion fluid, and oxygenation level of the perfusion fluid, wherein the system is configured to circulate the perfusion fluid between the organ chamber and the perfusion fluid container during system operation.

[0015] Perfusion fluid output is expressed as the perfusion fluid flow rate per unit volume per unit time. The term "organ compartment" refers to a compartment suitable for storing or culturing organ or tissue models.

[0016] Preferably, the system for storing or culturing organ or tissue models includes a system control device configured and programmed to acquire a measurement of at least one parameter selected from: the temperature of the perfusion fluid in the organ chamber, the temperature of the perfusion fluid in the perfusion fluid container, the glucose concentration in the perfusion fluid, the pH of the perfusion fluid, the flow parameters of the perfusion fluid including the pressure of the perfusion fluid expressed as a unit volume per unit time and / or the output of the perfusion fluid, the lactate concentration in the perfusion fluid, the sodium ion concentration in the perfusion fluid, and the chloride ion concentration in the perfusion fluid. The system control device is further configured and programmed to automatically and / or manually adjust at least one parameter selected from the perfusion fluid temperature in the organ chamber, the perfusion fluid temperature in the perfusion fluid container, the glucose concentration in the perfusion fluid, the pH of the perfusion fluid, the flow parameters of the perfusion fluid including the pressure and / or output of the perfusion fluid, the lactate concentration in the perfusion fluid, the sodium ion concentration in the perfusion fluid, the chloride ion concentration in the perfusion fluid, and the potassium ion concentration and oxygenation in the perfusion fluid, based on predetermined criteria.

[0017] Preferably, the system for storing or culturing organ or tissue models further includes means for measuring and controlling the oxygenation of the perfusion fluid and / or means for removing air from the perfusion fluid, wherein the system control means is preferably configured and programmed to obtain measurements of the oxygenation of the perfusion fluid and to automatically and / or manually adjust the oxygenation of the perfusion fluid based on predetermined criteria.

[0018] Preferably, the system for storing or culturing organ or tissue models further includes means for measuring the loss of the perfusion fluid, preferably based on a reading of the weight of the perfusion fluid removed from the system, and for replenishing the perfusion fluid, wherein the system control means is preferably configured and programmed to automatically and / or manually replenish the perfusion fluid based on predetermined criteria.

[0019] Preferably, the device for non-contact mixing of the infusion fluid is configured as an oscillating mechanism for oscillating the infusion fluid container, wherein the system control device is preferably configured and programmed to automatically and / or manually adjust the oscillation of the infusion fluid container.

[0020] Preferably, the oxygenation of the infusion fluid is performed using an oxygenator, which is externally connected to the infusion fluid container to form an oxygenator circuit, and the device for mixing the infusion fluid is configured in the form of a cylindrical design of the infusion fluid container, wherein during system operation, the infusion fluid is mixed by positioning the inlet and outlet of the oxygenator circuit on opposite sides of the infusion fluid container, tangent to the cylindrical wall of the infusion fluid container.

[0021] Preferably, the system includes means for obtaining a sample of the perfusion fluid.

[0022] Preferably, the organ chamber includes a rotation mechanism that allows the organ chamber to rotate during system operation, wherein the system control device is preferably configured and programmed to automatically and / or manually adjust the rotation of the organ chamber, wherein the system control device is preferably configured and programmed to rotate the organ chamber 180° every 30 minutes for at least two hours, or to rotate the organ chamber 90° every 15-60 minutes at least four times for at least two hours.

[0023] Preferably, the organ chamber and / or the perfusion fluid container and / or the device for measuring and controlling the oxygenation of the perfusion fluid have a device for adjusting the temperature in the range of 0 to 37°C.

[0024] Preferably, the system further includes an auxiliary device for metered addition of drugs and / or nutrients and / or infusion fluid components, wherein the system control device is preferably configured and programmed to automatically and / or manually metered add drugs and / or nutrients and / or infusion fluid components based on predetermined criteria.

[0025] Preferably, air is removed from the perfusion fluid by an air eliminator installed on the first line between the perfusion fluid container and the organ chamber, and during system operation, the movement of the perfusion fluid between the perfusion fluid container and the organ chamber in either direction is performed by at least one pump, preferably a peristaltic pump.

[0026] In a second aspect, the system according to the invention is provided for use in storing and / or culturing organ or tissue models, said organ or tissue models being harvested from a donor or prepared by other techniques, preferably using 3D bioprinting and / or 4D bioprinting and / or electrospinning.

[0027] Preferably, the system according to the invention is used to culture 3D bioprinted organs, wherein preferably, the rotating mechanism is used in the endothelial cell colonization vascular system stage.

[0028] Preferably, the disease status or development of the organ or tissue model is assessed during storage and / or culture.

[0029] Preferably, the effects of the bioactive substance on the disease state or development of the organ or tissue model are analyzed during storage and / or culture.

[0030] Preferably, the efficacy of drug treatment and / or gene therapy on the state of the organ or tissue model is evaluated during storage and / or culture.

[0031] Maintaining organ or tissue models at normal body temperature during culture is crucial, as it provides the organ or tissue model with optimal metabolic conditions corresponding to in vivo conditions, allowing for assessment of organ status.

[0032] The perfusion fluid used for culturing harvested organs contains, but is not limited to, concentrated red blood cells that tend to settle. The use of a non-contact device for mixing the perfusion fluid provides a uniform distribution of red blood cells within the fluid; this non-contact device preferably takes the form of a mechanism that agitates the perfusion fluid container or a container configuration that allows eddy currents to be generated within the container. Furthermore, it allows for maintaining the sterility of the flow system and reduces mechanical damage to the red blood cells that occurs when using other types of mixers. Mixing the perfusion fluid helps to uniformly heat the fluid and continuously mix the liquid components.

[0033] The rotation mechanism for rotating organ chambers is unrestricted for use in culturing organs prepared by 3D bioprinting technology and is used in the stage of endothelial cell colonization in bioprinted vascular systems. Preferably, the rotation mechanism rotates the organ chamber 180° every 30 minutes for at least two hours, or rotates the organ chamber 90° every 15-60 minutes in at least four separate sessions for at least two hours. The rotation mechanism enables colonization of the entire vascular system. Rotation is performed in a manner that allows cells to fall onto the channel wall under gravity and "adhere" to the channel wall. Rotation can be performed at small angles, for example, rotating 36° 10 times each time for at least two hours, or continuously rotating at a speed of, for example, one revolution per hour for at least two hours. The use of the rotation mechanism ensures a uniform distribution of colonized cells on the channel surface.

[0034] The system according to the invention can be used to cultivate and / or store and / or process (including regenerate) the following substances:

[0035] - Organs immediately harvested from deceased donors

[0036] - Organs harvested from living donors (e.g., family transplants or cross-transplants).

[0037] - Organs printed using 3D (and 4D) bioprinting techniques (organs with flow systems, including vascular systems).

[0038] - Tissue models with flow systems (including vascular systems) printed using 3D (and 4D) bioprinting technologies.

[0039] - Organ and tissue models prepared by electrospinning technology (organs with flow systems, including vascular systems).

[0040] - Organ and tissue models prepared by combining two methods (bioprinting and electrospinning)

[0041] Organs harvested from reproductive, laboratory, and genetically modified animals.

[0042] - Organs subjected to genetic manipulation

[0043] - Organs prepared under out-of-system (laboratory) conditions using tissue and genetic engineering techniques.

[0044] Organ and tissue models cultured in the system according to the invention can be used for medical and basic research as well as R&D work. Furthermore, the system according to the invention can be used to culture tissue models with flow systems (including vascular systems), which are prepared by 3D / 4D bioprinting or other techniques that allow for the preparation of models with flow systems (including electrospinning, which involves obtaining nanofibers from molten polymers or solutions thereof using an electric field), such as tissue models forming cancerous lesions, induced disease entities, and tissue models suitable for gene therapy research and personalized medicine.

[0045] This system can be used for storing and processing organs before transplantation, for testing the effects (i.e., toxicity and efficacy) of bioactive substances, for assessing the severity of a specific stage of disease, and for evaluating the effectiveness of drugs and gene therapies.

[0046] Advantageously, this system features precise dosing of drugs and active substances, as well as sampling for biological and chemical analysis. This makes it a versatile device that, in addition to storing / culturing and processing organs, allows for advanced research, preparation of organs for transplantation, and assessment of their functional status. Therefore, it will contribute to improving the effectiveness of transplantation procedures and facilitate advanced research (both basic and preclinical phases). Attached Figure Description

[0047] The invention is illustrated in the accompanying drawings, wherein... Figure 1 The diagram shows the overall structure of the system, which allows for the cultivation of organs harvested from donors, as well as organ or tissue models prepared using other techniques. Figure 2 The changes in perfusion parameters a) pressure [mm Hg], b) flow rate [ml / min], c) perfusion loss [g] and d) glucose concentration [mg / ml] during a 2-day porcine kidney culture in the system of Example 1 are shown. Figure 3 The perfusion parameters during porcine kidney culture in Example 1 are shown as: a) pH, b) Na. + c) Ion concentration [mmol / L], d) Glucose concentration [mg / dL], e) pCO2 [kPA], f) K + Ion concentration [mmol / L], f) saturation (oxygenation) [%], g) pO2 [kPa], h) Cl -Changes in ion concentration [mmol / L] and i) lactate concentration [mmol / L], Figure 4 The system for culturing pig kidneys from Example 1 is shown. Figure 5 A block diagram of the electronic system control device is shown. Figure 6 The geometry of the biomimetic pancreatic organ of Example 2 is shown. Figure 7 The figures show monitoring of a) heating-cooling plate temperature (bottom line) and organ compartment temperature (top line), in °C; b) arterial pressure RR [mm Hg]; and c) flow rate [ml / min]. Figure 8 This image shows a biomimetic pancreas obtained using a resonance method. Figure 9 The parameters of the perfusion fluid during 30 hours of biomimetic pancreas culture in Example 2 are shown as follows: a) pH, b) glucose concentration [mmol / L], c) lactate concentration [mmol / L], d) pO2 [kPa], e) Na+. + Ion concentration [mmol / L], f)K + Changes in ion concentration [mmol / L] Figure 10 The differences in oxygenation of the perfusion fluid after 30 hours are shown: Sample A—obtained after leaving the chamber containing the bionic organ—was a deoxygenated solution; Sample B—obtained before arriving at the chamber—was an oxygenated solution. Figure 11 The diagram illustrates the differences in insulin concentration after stimulation with glucose solutions of different concentrations, where gray lines mark glucose-stimulated islets on control inserts, and black lines mark islets printed in a bionic pancreas maintained in the system according to the invention. Figure 12 The system used in Example 2 for culturing a biomimetic pancreas obtained during 3D bioprinting is shown. Figure 13 A cross-section of the filling fluid container according to Example 3 is shown. Detailed Implementation

[0048] Example 1—Cultivation of pig kidneys

[0049] When optimizing the function of the system according to the invention, a pig kidney is used as a model. There are several practical reasons for using this organ:

[0050] -Easy to obtain and easy to collect, minimizing warm ischemia time.

[0051] -Easy to connect to vascularization

[0052] - High metabolism in organs allows for maximizing the system's potential.

[0053] - It can monitor organ function based on urine secretion.

[0054] Pig kidneys are harvested using a technique that interrupts meat production cycles. The animal is electrocuted, bled, and immediately transported to the operating room after the procedure to harvest the kidneys (thermal ischemia time <10 minutes). Harvesting is completed when the arteries are flushed with UW ice solution (University of Wisconsin solution). During the bleeding, the animal's blood is preserved aseptically (heparinized).

[0055] The composition of the perfusion fluid and the conditions for organ culture in the system according to the present invention are as follows:

[0056] The basic components of the perfusion solution used during culture under normal body temperature conditions are a low-potassium solution (Belzer) used for mechanical perfusion. The MPS fluid was prepared using a UW machine perfusion solution enriched with erythrocyte concentrate at a 7:3 ratio, allowing for a hematocrit of approximately 0.15. Additionally, the MPS fluid was supplemented with CaCl2, amino acid concentrate (Trimel N9-1070 EC), vitamins, sodium bicarbonate (to achieve a pH between 7.35 and 7.45), heparin, antibiotics (PenStrep), insulin, and dexamethasone. The mean perfusion pressure was compared at 50 mmHg and 75 mmHg. The partial pressure of oxygen in the arterial branches of the system was approximately 50 mmHg, flowing through the oxygenator at approximately 3 L / min. Kidneys were cultured for 48 hours before histopathological evaluation. Glucose concentrations were maintained at 100–150 mg% by continuous infusion based on the perfusion fluid concentration. Losses due to diuresis were automatically replenished with Ringer's solution (at a 1:1 ratio). Viability of the cultured kidneys was assessed based on urine production, glucose consumption, and blood saturation (oxygenation) upstream and downstream of the organ. If reduced perfusion is caused by increased vascular resistance, the kidneys are treated with urapidil (Ebrantil) infusion (as needed). Figure 2 and Figure 3 The changes in perfusion fluid parameters during porcine kidney culture in Example 1 are shown.

[0057] Systems for culturing pig kidneys ( Figure 4 This includes the following:

[0058] - A sealed organ chamber 2, equipped with means for regulating the temperature of the perfusion fluid located in chamber 2 during system operation. Chamber 2 includes a heating-cooling circuit 13 in the form of a heating jacket. Chamber 2 is equipped with ports that allow connection of organs, allow infusion fluid inflow, and allow infusion fluid outflow, as well as a bubble trap—bubble eliminator 6 designed to change the pressure or volume of the gas contained in chamber 2.

[0059] - An infusion container 1 is equipped with a device for regulating the temperature of the infusion fluid in the form of a heating-cooling plate 11, the temperature of which is regulated by a Peltier module 27. The infusion container 1 is connected to the chamber 2 via at least one first line 17 and at least one second line 18. During operation, the infusion fluid can flow from the infusion container 1 to the organ chamber 2 via the first line 17, and during operation, the infusion fluid can flow from the organ chamber 2 to the infusion container 1 via the second line 18.

[0060] - An apparatus for measuring and controlling the oxygenation of the infusion fluid, comprising an oxygenation device: an oxygenator 5, an oxygenation-saturation sensor 19, a gas mixture cylinder assembly, and a pressure reducer, which provides appropriate pressure and gas flow through the oxygenator. The oxygenator 5 also includes a temperature sensor 32 for the oxygenator 5 and a heating-cooling circuit 12 in the form of a heating jacket for the oxygenator 5. The apparatus for measuring and controlling the oxygenation of the infusion fluid is also referred to as the circuit of the oxygenator 5.

[0061] The temperature of the perfusion fluid is regulated by a heating-cooling circuit 12 and 13 of the organ chamber 2 and oxygenator 5 that allows the flow of heating or cooling fluid, a heating-cooling circuit of the perfusion fluid container 1, a temperature sensor 30 of the organ chamber 2, a temperature sensor 31 of the perfusion fluid container 1, and a temperature sensor 32 of the oxygenator 5, a regulating system that provides the flow of heating or cooling fluid, preferably distilled water, and a pump 9, wherein the heating or cooling fluid does not come into contact with the organ or perfusion fluid and is only used to transfer heat from the heating device to the heating jacket of the organ chamber 2 and the heating jacket of the oxygenator 5.

[0062] - A device 4 for measuring glucose concentration in the perfusion fluid and for metering glucose into the perfusion fluid includes an enzyme electrode-based sensor 25, an electronic measurement system, and a pump 9 for metering glucose-containing replenishment solution. The enzyme electrode used is a disposable device specifically designed for measurement in a flow system and in direct contact with the perfusion fluid. The glucose concentration data is displayed on a user panel, and based on this reading, glucose-containing replenishment solution is metered and added automatically or as needed by the operator via system control.

[0063] -A device 3 for measuring the pH of an irrigation solution and for metering the addition of a substance for adjusting the pH of the irrigation solution, comprising a pH electrode, an electronic measuring system, and a pump for metering the addition of the substance for adjusting the pH.

[0064] - A device for non-contact mixing of the filling fluid, provided in the form of a swing mechanism 16 for the filling fluid container 1. The swing mechanism 16 causes the filling fluid container 1 to swing.

[0065] - A device for removing air from the perfusion fluid includes two perfusion fluid air bubble traps—air bubble eliminators 6. One air bubble trap is located in the circuit of the oxygenator 5, and the other air bubble trap (mentioned above) is located directly in front of the organ chamber 2.

[0066] - A device for measuring and controlling the flow parameters (including the perfusion fluid pressure) of the perfusion fluid flowing from the perfusion fluid container 1 into the organ chamber 2 during system operation. The device for measuring and controlling the flow parameters of the perfusion fluid includes an electromechanical membrane pressure sensor 7, an electronic control system, and a peristaltic pump 9. Pressure measurement is performed non-contactly. The perfusion fluid is separated from the sensor 7 by a flexible membrane. The perfusion pressure is set by the perfusion operator using the system control device on a user panel. The system control device automatically selects the flow rate to achieve the set pressure. A maximum permissible flow rate is also preset on the user panel.

[0067] - A device 14 for measuring the loss and replenishment of the infusion fluid includes a weight-based system based on a tension sensor, an electronic measurement system, and a metering pump 9. The amount of fluid added is displayed on the operator panel and, depending on the settings, can be automatically or manually added by the operator using the system control device.

[0068] - A system control device configured and programmed to acquire a measurement value of at least one parameter selected from: the temperature of the perfusion fluid in organ chamber 2, the temperature of the perfusion fluid in perfusion fluid container 1, the concentration of glucose in the perfusion fluid, the pH of the perfusion fluid, the flow parameters of the perfusion fluid expressed per unit volume per unit time, including the pressure and / or output of the perfusion fluid, the concentration of lactate in the perfusion fluid, the concentration of sodium ions in the perfusion fluid, the concentration of chloride ions in the perfusion fluid, the concentration of potassium ions in the perfusion fluid, and the oxygenation of the perfusion fluid, and to automatically and / or manually adjust based on a predetermined standard for at least one parameter selected from: the temperature of the perfusion fluid in organ chamber 2, the temperature of the perfusion fluid in perfusion fluid container 1, the concentration of glucose in the perfusion fluid, the pH of the perfusion fluid, the flow parameters of the perfusion fluid including the pressure and / or output of the perfusion fluid, the concentration of lactate in the perfusion fluid, the concentration of sodium ions in the perfusion fluid, the concentration of chloride ions in the perfusion fluid, the concentration of potassium ions in the perfusion fluid, and the oxygenation of the perfusion fluid.

[0069] The main controller of the system control device consists of two parts: the first part is based on a microprocessor 21, which ensures real-time operation by controlling all peripheral devices of the device. The second processor 20 is responsible for the graphical user interface (GUI), enabling the observation and input of parameters.

[0070] The TEC controller 23 is designed to control the Peltier modules 27 so that they maintain the infusion fluid temperature between 0 and 37°C, thereby allowing organ processing under normal and low body temperature conditions. The fan 28 is used to control the radiator temperature, ensuring that the temperature difference between the hot and cold sides of the Peltier modules does not exceed 20°C.

[0071] The controller 22 records the following temperature measurements: the Peltier module 27, the infusion fluid heating-cooling plate 11, the water jacket of the organ chamber 2, and the infusion fluid container 1.

[0072] Several peristaltic pumps 9 are used, the first of which is used for the main perfusion fluid circulation and is controlled to maintain a constant hydraulic pressure controlled by a tension pressure sensor 7. The other three pumps 9 are used for drug delivery. Drug delivery can be performed at a specific rate of ml / min.

[0073] The control panel of the system according to the present invention can be accessed at the web browser level, making it accessible from any device, such as a mobile phone, tablet, or laptop, as long as it has internet access.

[0074] The control panel allows setting the following parameters: temperature of the heating-cooling plate 11, parameters of the PID controller for temperature control, turning pressure control on and off, preset perfusion fluid pressure [mmHg], minimum and maximum flow (output) of the perfusion fluid, preset speed of each stepper motor 29 [ml / min], turning on the perfusion fluid mixing mechanism, and setting the perfusion fluid mixing speed. The control panel also allows monitoring of measured parameters such as the pH of the perfusion fluid, the glucose concentration in the perfusion fluid, and the temperature of the heating-cooling plate 11. Figure 5 This is a block diagram of the electronic system control device.

[0075] Example 2—Cultivation of organs obtained through bioprinting technology.

[0076] Experiments using a biomimetic pancreatic organ were conducted using 3D bioprinting. The organ comprises a scaffold containing the islets of Langerhans made from basic bio-ink and a duct system made from vascular bio-ink. Figure 6 The ductal system consists of a main vessel, branching into three spirally arranged secondary vessels that merge into a single draining vessel. Description of the organ's geometric parameters:

[0077] The overall external dimensions of the model are 32×40×17.5mm.

[0078] - The length of a single secondary blood vessel is 340mm.

[0079] - The total length of secondary vessels is 1020mm

[0080] - The diameter of the main blood vessel is 1.5 mm.

[0081] - Secondary blood vessels have a diameter of 1 mm.

[0082] - The volume of the pancreatic islet stent is 20.5 ml.

[0083] - The volume of the catheter is approximately 0.9 ml.

[0084] The culture process was conducted under controlled temperature conditions: the temperature in organ chamber 2 was between 37 and 39°C; the temperature of the heating-cooling plate 11 used to heat the liquid was between 36.5 and 37°C. Flow and pressure levels were also monitored throughout the experiment. Figure 7 ).

[0085] The method of connecting the bionic organ to organ chamber 2 and monitoring parameters (pressure, flow, temperature) does not affect the structure and function of the vascular system printed in the bionic organ. This was confirmed by the resonance method. Figure 8 ).

[0086] During the incubation of the biomimetic organ in organ chamber 2, the following parameters of the perfusion fluid were monitored in real time: oxygenation, pH, glucose concentration, lactate concentration, sodium ions, and potassium ions. Figure 9 ).

[0087] The control of parameters allows for monitoring of the perfusion fluid composition and, if necessary, improvement of its quality, enabling the maintenance of the biomimetic organ under optimal culture / incubation conditions. Furthermore, red blood cells were added to the perfusion fluid used in this system as biological oxygen carriers. Perfusion fluid samples were obtained during the experiment (after 30 hours), showing significant differences in oxygenation between the perfusion fluids. Figure 10 Sample A – obtained after leaving chamber 2 where the bionic organ is located – deoxygenated solution; Sample B – obtained before reaching chamber 2 – oxygenated solution.

[0088] In addition, the bionic organ was tested to evaluate its function. Besides controlling the aforementioned basic parameters, this included sampling at set time points to assess insulin concentrations after stimulation with glucose solutions of different concentrations. Figure 11 There was no difference in biomimetic organ results between islets printed as biomimetic organs and islets that did not undergo the 3D bioprinting process.

[0089] Systems for cultivating bionic organs Figure 12 This includes the following:

[0090] - A sealed, movable organ chamber 2, equipped with means for regulating the temperature of the perfusion fluid located in chamber 2 during device operation. Chamber 2 includes a heating-cooling circuit 13 in the form of a heating jacket, a rotation mechanism 15 for rotating chamber 2, and a rotation drive for chamber 2. Chamber 2 is equipped with ports that allow connection of organs, allow infusion fluid inflow, allow infusion fluid outflow, and a bubble eliminator 6, which is designed to change the pressure or volume of the gas contained in chamber 2.

[0091] - Infusion fluid container 1, the infusion fluid container 1 is equipped with a device for regulating the temperature of the infusion fluid in the form of a heating-cooling plate 11, the temperature of which is regulated by a Peltier module 27, wherein the infusion fluid container 1 is connected to the chamber 2 via at least one first line 17 and at least one second line 18, during operation the infusion fluid can flow from the infusion fluid container 1 to the organ chamber 2 through the first line 17, and during operation the infusion fluid can flow from the organ chamber 2 to the infusion fluid container 1 through the second line 18.

[0092] The temperature of the perfusion fluid is regulated by a heating-cooling circuit of organ chamber 2 and perfusion fluid container 1, a temperature sensor 30 of organ chamber 2, a temperature sensor 31 of perfusion fluid container 1, a control system, and a pump 9 that provides the flow of heating or cooling fluid. Organ chamber 2 has a heating-cooling circuit 13 that allows the flow of heating or cooling fluid.

[0093] - A device 4 for measuring the glucose concentration in the perfusion fluid and adding glucose to the perfusion fluid in a metered manner includes an enzyme electrode-based sensor 25, an electronic measurement system, and a pump 9 for metered addition of glucose-containing liquid. The enzyme electrode used is a disposable device specifically designed for measurement in a flow system with direct contact with the perfusion fluid. The glucose concentration data is displayed on a user panel, and the system control unit automatically adds replenishment fluid in metered manner based on this reading or when required by the operator.

[0094] - A device 3 for measuring the pH of the irrigation fluid and for metering the addition of a substance for adjusting the pH of the irrigation fluid, comprising a pH electrode, an electronic measuring system, and a pump 9 for metering the addition of the substance for adjusting the pH.

[0095] - A device for non-contact mixing of the filling fluid, provided in the form of a swing mechanism 16 for the filling fluid container 1. The swing mechanism 16 causes the filling fluid container 1 to swing.

[0096] - A device for removing air from the perfusion fluid, including a perfusion fluid bubble trap—a bubble eliminator 6 located directly in front of the organ chamber 2.

[0097] - A device for measuring and controlling the flow parameters (including the pressure of the perfusion fluid) of the perfusion fluid flowing from the perfusion fluid container 1 into the organ chamber 2 during system operation. The device for measuring and controlling the flow parameters of the perfusion fluid includes an electromechanical membrane pressure sensor 7, an electronic control system, and a peristaltic pump 9. Pressure measurement is performed non-contactly. The perfusion fluid is separated from the sensor 7 by a flexible membrane. The perfusion pressure is set by the perfusion operator using the system control device on a user panel. The system control device automatically selects the flow rate to achieve the set pressure. A maximum permissible flow rate is also preset on the user panel.

[0098] - The system control device is compatible with that in Embodiment 1. This system control device also allows for control of the rotation of organ chamber 2.

[0099] Example 3

[0100] To culture the porcine kidneys of Example 1, a system corresponding to that of Example 1 was used, wherein the perfusion container 1 was not equipped with the oscillation mechanism 16 and was heated using the Peltier module 27. Container 1 was cylindrical, with the port located at its lower part ( Figure 13 Non-contact mixing of the infusion fluid is achieved by setting the inlet and outlet of the infusion fluid in the oxygenator 5 circuit on opposite sides of the infusion fluid container 1, tangential to its cylindrical wall, thus introducing vortex motion within the container 1. The liquid flow rate output at the inlet and outlet of the oxygenator 5 circuit is set between 0 and 2 L / min. Heating or cooling of the infusion fluid is achieved using a water jacket 33. The infusion fluid container 1 is fabricated using 3D printing technology.

[0101] List of reference numerals in the attached figures: 1-Perfusion fluid container; 2-Organ compartment; 3-Device for measuring pH; 4-Device for measuring glucose concentration; 5-Oxygenator; 6-Bubble eliminator; 7-Pressure sensor for perfusion fluid; 8-Device for obtaining perfusion fluid samples; 9-Peristaltic pump; 10-Container for drugs and / or nutrients and / or perfusion fluid components; 11-Heating-cooling plate; 12-Heating-cooling circuit of the oxygenator; 13-Heating-cooling circuit of the organ compartment; 14-Device for measuring perfusion fluid loss; 15-Rotation mechanism for rotating the organ compartment; 16-Oscillating mechanism of the perfusion container; 17-First pipeline; 18-Second pipeline; 19-Oxygenation / saturation sensor; 20-GUI processor; 21-Real-time microprocessor; 22-TEC controller; 23-Stepper motor controller; 24-Temperature measurement system; 25-Glucose sensor; 26-Level sensor; 27-Peltier module; 28-Fan; 29-Stepper motor; 30-Temperature sensor of organ chamber; 31-Temperature sensor of perfusion container; 32-Temperature sensor of oxygenator; 33-Water jacket of perfusion container of Example 3.

Claims

1. A system for storing or culturing organ or tissue models, said organ or tissue models being harvested from a donor and selected from pancreas, liver, kidney, lung, heart, small intestine, large intestine, thyroid gland, skin, brain, or prepared using other techniques, using 3D bioprinting and / or 4D bioprinting and / or electrospinning techniques, in the form of organ and / or tissue models printed with flow systems or vascular systems, said system comprising: - A chamber (2) for an organ or tissue model, said chamber (2) being equipped with means for measuring and regulating the temperature of the perfusion fluid placed in said chamber (2) during operation of said system, wherein said means for measuring and regulating the temperature of said perfusion fluid includes a water jacket, - An infusion container (1) equipped with means for measuring and regulating the temperature of the infusion fluid in the container (1) during system operation, including a heating-cooling plate (11), wherein the infusion container (1) is connected to the organ chamber (2) via at least one first line (17) and at least one second line (18), wherein during operation the infusion fluid can flow from the infusion container (1) to the organ chamber (2) via the first line (17), and during operation the infusion fluid can flow from the organ chamber (2) to the infusion container (1) via the second line (18). -A device (4) for measuring the glucose concentration in the perfusion solution and for adding glucose to the perfusion solution in a metered manner. -A device (3) for measuring the pH of the infusion solution and for adding, in metered amounts, a substance for adjusting the pH of the infusion solution. - A device for non-contact mixing of the infusion fluid, - A device for measuring and controlling the flow parameters of the infusion fluid, the flow parameters including infusion fluid pressure and / or infusion fluid output expressed as a unit volume per unit time. - Optionally, an apparatus for measuring and controlling one or more parameters selected from: lactate concentration in the perfusion fluid, sodium ion concentration in the perfusion fluid, chloride ion concentration in the perfusion fluid, potassium ion concentration in the perfusion fluid, and oxygenation level of the perfusion fluid, wherein the system is configured to circulate the perfusion fluid between the organ chamber (2) and the perfusion fluid container (1) during system operation. The device for non-contact mixing of the infusion fluid is configured in the form of a cylindrical design of the infusion fluid container (1), wherein during system operation, the infusion fluid is mixed by setting the inlet and outlet of the oxygenator (5) circuit in the infusion fluid container (1) on opposite sides of the infusion fluid container (1) and tangential to the cylindrical wall of the infusion fluid container (1).

2. The system for storing or culturing organ or tissue models according to claim 1, comprising a system control device configured and programmed to obtain a measurement of at least one parameter selected from: the temperature of the perfusion fluid in the organ chamber (2), the temperature of the perfusion fluid in the perfusion fluid container (1), the glucose concentration in the perfusion fluid, the pH of the perfusion fluid, the flow parameters of the perfusion fluid including the pressure of the perfusion fluid expressed in unit volume per unit time and / or the output of the perfusion fluid, the lactate concentration in the perfusion fluid, the sodium ion concentration in the perfusion fluid, the chloride ion concentration in the perfusion fluid, and the potassium ion concentration in the perfusion fluid, and the oxygenation level of the perfusion fluid, wherein the system control device is further configured and programmed to perform automatic and / or manual adjustment based on predetermined criteria of the at least one parameter.

3. The system for storing or culturing organ or tissue models according to claim 2, further comprising means for measuring and controlling the oxygenation of the perfusion fluid and / or means for removing air from the perfusion fluid, wherein the system control means is configured and programmed to obtain oxygenation measurements of the perfusion fluid and to automatically and / or manually adjust the oxygenation of the perfusion fluid based on predetermined criteria.

4. The system for storing or culturing an organ or tissue model according to any one of claims 1 to 3, further comprising means (14) for measuring the loss of the perfusion fluid based on a reading of the weight of the perfusion fluid removed from the system and replenishing the perfusion fluid, wherein the system control means is configured and programmed to automatically and / or manually replenish the perfusion fluid based on predetermined criteria.

5. The system for storing or culturing organ or tissue models according to claim 1, wherein oxygenation of the perfusion fluid is performed using an oxygenator (5) externally connected to the perfusion fluid container (1) to form the oxygenator (5) circuit.

6. The system for storing or culturing an organ or tissue model according to any one of claims 1 to 3, comprising means (8) for obtaining a sample of the perfusion fluid.

7. The system for storing or culturing organ or tissue models according to any one of claims 1 to 3, wherein the organ chamber (2) includes a rotation mechanism (14) that allows the organ chamber (2) to rotate during system operation, wherein the system control device is configured and programmed to automatically and / or manually adjust the rotation of the organ chamber (2), wherein the system control device is configured and programmed to rotate the organ chamber (2) by 180° every 30 minutes for at least two hours, or to rotate the organ chamber (2) by 90° every 15-60 minutes at least four times for at least two hours.

8. The system for storing or culturing organ or tissue models according to any one of claims 1 to 3, wherein the organ chamber (2) and / or the perfusion container (1) and / or the device for measuring and controlling the oxygenation of the perfusion has a means for adjusting the temperature in the range of 0 to 37°C.

9. The system for storing or culturing organ or tissue models according to any one of claims 1 to 3, further comprising an additional device for metered addition of drugs and / or nutrients and / or perfusion fluid components, wherein the system control device is configured and programmed to automatically and / or manually metered add drugs and / or nutrients and / or perfusion fluid components based on predetermined criteria.

10. The system for storing or culturing organ or tissue models according to claim 3, wherein air is removed from the perfusion fluid by means of an air eliminator (6) installed on the first line (17) between the perfusion fluid container (1) and the organ chamber (2), and during operation of the system, movement of the perfusion fluid in either direction between the perfusion fluid container (1) and the organ chamber (2) is performed by means of at least one pump (9), a peristaltic pump.

11. The use of the system as defined in any one of claims 1 to 3 for storing and / or culturing organ or tissue models, said organ or tissue models being harvested from a donor or prepared by other techniques, utilizing 3D bioprinting and / or 4D bioprinting and / or electrospinning.

12. The use according to claim 11, wherein the system of claim 7 is used for culturing 3D bioprinted organs, wherein, The rotating mechanism (14) is used in the endothelial cell colonization vascular system stage.

13. The use according to claim 11, wherein the state or development of disease in an organ or tissue model is assessed during storage and / or culture.

14. The use according to claim 13, wherein the effect of the bioactive substance on the state or development of disease in an organ or tissue model is analyzed during storage and / or culture.

15. The use according to claim 13, wherein the efficacy of drug treatment and / or gene therapy on the state of the organ or tissue model is evaluated during storage and / or culture.

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