Dynamic organ preservation system

Abstract

The dynamic organ preservation system includes an insulated container with a lid that seals an internal volume. Inside the internal volume is an organ container with a lid that contains an organ immersed in a perfusion fluid, the lid having at least one inlet port and at least one outlet port. At least one pressure bag for further storing the perfusion fluid is inclined at a certain angle above the organ container. At least one infusion line enters the organ container through at least one inlet port and connects at least one pressure bag to the organ, allowing the perfusion fluid to flow into the organ by gravity. A sampling line connected to the organ exits the organ container through at least one outlet port so that a user can monitor the organ's condition. At least one cooling mechanism maintains the organ at hypothermia for a certain period of time without requiring electricity.

Description

Detailed Description of the Invention

[0001] (Related Applications) This application claims the benefit of U.S. Provisional Application No. 63 / 606,000 filed on December 13, 2023, entitled "Dynamic Organ Preservation System", and U.S. Provisional Application No. 63 / 538,471 filed on September 14, 2023, entitled "Dynamic Organ Preservation". These provisional applications are hereby incorporated herein by reference in their entirety for all purposes. TECHNICAL FIELD

[0002] The present disclosure relates generally to extracorporeal organ support, and more specifically, to a dynamic organ preservation system that enables cold preservation of extracorporeal organs with perfusion and monitoring functions without requiring an external power source during operation. BACKGROUND ART

[0003] Organ transplantation is the only curative treatment for many end-stage diseases. The shortage of viable donor organs for transplantation is exacerbated by significant bottlenecks arising from inadequate organ preservation, recovery, and transport strategies when donor organs become available. Static cold preservation is a common method of organ preservation, but the use of static cold preservation is limited by many problems, including tissue damage induced by prolonged hypothermic preservation, difficulty in assessing the function and viability of donor organs, the inevitability of ischemia-reperfusion injury, and limited opportunities for organ repair. As an alternative, mechanical perfusion systems are expected to revolutionize organ preservation by enabling more advanced organ perfusion and being equipped with monitoring functions. However, mechanical perfusion systems are expensive, bulky, heavy, require significant external power, and may include many or all of the other limitations that static cold preservation has. SUMMARY OF THE INVENTION PROBLEMS TO BE SOLVED BY THE INVENTION

[0004] This specification describes a dynamic organ preservation system that enables the refrigerated storage of extracorporeal organs for longer periods and in a more stable state than conventional preservation mechanisms. This dynamic organ preservation system does not require an external power source during operation and is equipped with perfusion and monitoring functions (e.g., ensuring temperature stability). Furthermore, the dynamic organ preservation system is highly portable, significantly reducing the cost of preserving and / or transporting extracorporeal organs that may be life-saving. [Means for solving the problem]

[0005] In one embodiment, the disclosure includes an organ preservation system for dynamically preserving an extracorporeal organ. The system may include an insulated container having a fixable lid for sealing the inner volume of the insulated container. The inner volume of the insulated container may further house an organ container, at least one pressure bag, at least one infusion line, a sampling line, and at least one cooling mechanism. The organ container may house an organ at least partially immersed in a certain volume of perfusion fluid and may have a lid for completely sealing the organ container. The lid may include at least one inlet port and at least one outlet port. At least one pressure bag may store another volume of perfusion fluid, is positioned above the organ container, and may be inclined at an angle to the lid of the organ container. At least one infusion line may enter the organ container through at least one inlet port and connect at least one pressure bag to the organ, allowing perfusion fluid to flow from at least one pressure bag to the organ by gravity. A sampling line may be connected to the organ, allowing a user to monitor the organ's condition. The sampling line may exit the organ container through at least one outlet port. At least one cooling mechanism can maintain a hypothermic state in organs for a certain period of time without requiring electricity.

[0006] In another embodiment, the disclosure may include a method for preserving a hypothermic organ by dynamically preserving an extracorporeal organ. The steps include: placing the organ in an organ container such that the organ is at least partially immersed in a certain volume of perfusion fluid, the organ container being located in the inner volume section of an insulated container; connecting at least one infusion line and a sampling line to the organ; sealing the organ container with a lid having at least one inlet port for receiving at least one infusion line and at least one outlet port for receiving a sampling line; connecting at least one infusion line to at least one pressure bag that places the organ container in an insulated container having an inner volume section, a fixable lid for sealing the inner volume section, and at least one cooling mechanism located within the inner volume section; placing at least one pressure bag storing another volume of perfusion fluid above the organ container in the insulated container, tilted at an angle to the lid of the organ container, so that another volume of perfusion fluid flows into the organ by gravity; cooling the inner volume section of the insulated container using one or more cooling mechanisms to maintain the organ at hypothermia for a certain period of time without requiring electricity. The organ is perfused for a set period of time without requiring power, using a perfusion fluid supplied by gravity from at least one pressure bag to the organ via at least one infusion line, where at least one infusion line is cannulated into at least one vein and / or artery of the organ. The user can monitor the state of the organ using at least one sensor accessible by a sampling line, with a lockable lid closed.

[0007] The features described above and other features of this disclosure will become apparent to those skilled in the art in which this disclosure relates by reading the following description with reference to the accompanying drawings. [Brief explanation of the drawing]

[0008] [Figure 1] This is a schematic diagram of an example of a dynamic organ preservation system. [Figure 2] This is a schematic diagram of an example of a dynamic organ preservation system. [Figure 3]Figure 2 is a schematic diagram of the system including the dynamic organ preservation system and external devices. [Figure 4] This figure shows the setup and use of an exemplary dynamic organ preservation system, similar to those shown in Figures 1 and 2. [Figure 5] This figure shows the setup and use of an exemplary dynamic organ preservation system, similar to those shown in Figures 1 and 2. [Figure 6] This figure shows the setup and use of an exemplary dynamic organ preservation system, similar to those shown in Figures 1 and 2. [Figure 7] This figure shows the setup and use of an exemplary dynamic organ preservation system, similar to those shown in Figures 1 and 2. [Figure 8] This figure shows the setup and use of an exemplary dynamic organ preservation system, similar to those shown in Figures 1 and 2. [Figure 9] This figure shows the setup and use of an exemplary dynamic organ preservation system, similar to those shown in Figures 1 and 2. [Figure 10] This figure shows the setup and use of an exemplary dynamic organ preservation system, similar to those shown in Figures 1 and 2. [Figure 11] This figure shows an example of the configuration of an organ container. [Figure 12] Figures 1 and 2 show a graphical representation of the temperatures of an exemplary organ and adiabatic container (registered trademark: Yeti) over a 12-hour period in a dynamic organ preservation system similar to those shown. [Figure 13] This figure shows an example of a pressure bag. [Figure 14] This figure shows an example of an insertion port. [Figure 15] Figures 1 and 2 show kidney containers for use in a dynamic organ preservation system. [Figure 16] Figures 1 and 2 show kidney containers for use in a dynamic organ preservation system. [Figure 17]This is a diagram of cannulation into the heart for use in the system described above. [Figure 18] This is a diagram of cannulation into the heart for use in the system described above. [Figure 19] This is a photographic diagram showing examples of different cannulas that may be used by one or more of the systems described above. [Figure 20] This is a photographic diagram showing examples of different cannulas that may be used by one or more of the systems described above. [Figure 21] This is a photographic diagram showing examples of different cannulas that may be used by one or more of the systems described above. [Figure 22] This is a process flowchart showing a method for hypothermically preserving an organ using a dynamic organ preservation system. [Figure 23] This shows a schematic diagram of an experiment and a photographic diagram of an example system setup. [Figure 24] This is a graphical representation of experimental results. [Figure 25] This is a graphical representation of experimental results. [Figure 26] This is a graphical representation of experimental results. [Figure 27] This is a graphical representation of experimental results. [Figure 28] This is a photograph and graphical representation of experimental results. [Figure 29] This is a photograph and graphical representation of experimental results. [Mode for Carrying Out the Invention]

[0009] Definitions Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present disclosure belongs.

[0010] As used herein, the singular forms "a", "an" and "the" may include the plural forms as well, unless the context clearly indicates otherwise.

[0011] As used herein, the terms “includes” and / or “contains” may identify the presence of the described features, steps, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, steps, operations, elements, components, and / or combinations thereof.

[0012] As used herein, the term "and / or" may include any combination of one or more of the related enumerated items.

[0013] As used herein, terms such as “first,” “second,” etc., should not be used to limit the elements described by these terms. These terms are used solely to distinguish one element from another. Thus, the “first” element discussed below may also be called the “second” element without departing from the teachings of this disclosure. The order of operations (or actions / steps) is not limited to the order shown in the claims or drawings unless otherwise specified.

[0014] As used herein, terms such as “dynamic organ preservation” refer to techniques that enable the preservation of organs outside the body with perfusion and monitoring capabilities. In a dynamic organ preservation system, the perfusion fluid flows through the organ, oxygenating it to maintain it in or near physiological stability, and monitoring one or more health factors of the organ.

[0015] As used herein, the term “perfusion fluid” refers to any fluid circulating within an extracorporeal organ to help maintain the organ’s physiological stability. Perfusion fluid may be blood and / or artificial solutions. Perfusion fluid may contain one or more added nutrients and / or added oxygen. In some examples, perfusion fluid may be liquid, and / or primarily liquid.

[0016] As used herein, the term “pressurized bag” refers to a specialized container designed to use air pressure to increase the infusion rate of a fixed volume of liquid (e.g., perfusion fluid) stored within the container into an infusion line and organs.

[0017] As used herein, the term “infusion line” refers to a pathway through which a perfusion fluid (e.g., blood, artificial perfusion fluid, etc.) flows from a pressure bag to an organ in order to supply oxygen and nutrients.

[0018] As used herein, the term “effluent” refers to waste fluid from organs and / or organ containers (e.g., used perfusion fluid).

[0019] As used herein, the term “(organ) condition” refers to the condition of an organ relating to its appearance, quality, function, etc. Organ condition can be determined based on one or more measured biomarkers. Examples include, but are not limited to, venous oxygen saturation (SvO2) and arterial oxygen saturation (SaO2) (inflow and outflow saturation), lactate concentration, mitochondrial markers, glucose concentration, pH, hematocrit and / or hemoglobin (Hgb) levels, organ-specific markers such as troponin for the heart, organ-specific exosome markers, temperature (inflow, outflow, and / or container), organ-specific metabolites, inflow pressure of perfusion fluid, line pressure of infusion lines, and organ-specific venous and / or arterial blood flow (e.g., measured in the portal vein and / or hepatic artery of the liver), depending on the type of organ.

[0020] As used herein, the term “hypothermia” refers to an ambient or organ temperature of at least 32°C, preferably about 0°C to 14°C. Lowering the temperature of a normal organ to a temperature lower than that normally maintained by organ homeostasis reduces the metabolic rate of the organ, and thus reduces the demand for oxygen and substrates that the tissue needs to protect itself from the effects of ischemia and hypoxia.

[0021] As used herein, the term “living organism” can refer to any warm-blooded organism, including but not limited to humans, pigs, rats, mice, dogs, cats, goats, sheep, horses, monkeys, apes, rabbits, and cattle.

[0022] As used herein, the terms “extracorporeal organ” or “extracorporeal organ” refer to any organ located outside the body of an organism (e.g., an organ provided by an organ donor, an organ cultured in a laboratory, or an organ isolated (voluntarily or involuntarily) from a host organism). Extracorporeal organs may include, but are not limited to, internal organs (e.g., heart, liver, lungs, kidneys, pancreas, small intestine, intestine, etc.), external organs (e.g., skin), tissues, bioengineered grafts, xenografts, or limbs (e.g., arms, legs, hands, feet, fingers, toes, etc.).

[0023] overview Donor organs begin to lose function after the donor's death or immediately after being removed from a living donor's body. Preservation can extend the function of donor organs for transplantation into the recipient's body. Traditionally, donor organs have been preserved via static refrigeration or mechanical perfusion systems, both of which have significant limitations in terms of preservation duration, transportability, and / or quality of preservation. Static refrigeration, the most common method of preserving solid organs, refers to cooling the organ to a hypothermic state and maintaining it for a certain period to slow metabolic processes and degradation. The time limit for static refrigeration is organ-dependent, but is generally limited to about 4-8 hours before ischemia or other irreversible damage occurs. In fact, the combination of ischemia and static refrigeration is linked to an increased risk of post-transplant complications such as graft failure, graft failure, and ischemic biliary injury. Mechanical perfusion systems, either hypothermic or normothermic, continuously or pulsatilely deliver perfusion fluid through the organ's vascular system, which can extend the organ's preservation time by up to approximately 10 hours in some cases. However, mechanical perfusion systems are traditionally heavy, bulky, and extremely difficult to transport due to size limitations and external power requirements for operating all system components. For example, a mechanical perfusion system may include a rotor pump or centrifugal pump, oxygen supply unit, heat exchanger, and bulky monitoring system. The cost per transplanted organ is very high, reaching approximately $25,000 for disposable static refrigeration and around $95,000 for a mechanical perfusion system. Furthermore, transporting a mechanical perfusion system requires specialized transport means (aircraft or automobile) that can withstand its weight and have sufficient power supply capacity and space for the system and technicians.

[0024] The time, logistics, and cost of transporting organs to where they are needed are the biggest obstacles to the success of transplantation using available donor organs. Dynamic organ preservation systems, such as those described herein, which do not require an external power source during operation, can significantly improve each of these obstacles. By using dynamic organ preservation systems, the time for which organs can be safely and effectively preserved can be increased. Furthermore, dynamic organ preservation systems can be easily transported by being at least (1) relatively small and (2) not requiring an external power source during operation. Dynamic cryopreservation systems can uniformly cool organs to a desired hypothermia and maintain that temperature for at least 12 or 24 hours, while simultaneously perfusing the organs using pressure and gravity supply systems. In some examples, sensors (e.g., with reusable built-in batteries) can be connected to external devices to monitor the condition of the organs and sample effluent in real time.

[0025] system One aspect of the present disclosure may include a system 100 (Figure 1) capable of preserving organs at hypothermia for a certain period of time while perfusing them. The system 100 is easily transportable and its size, shape, and weight may be adjusted to accommodate any type and / or number of organs. As shown in Figure 1, the dynamic organ preservation system 100 may include an insulated container 102 (e.g., an insulated container) having an internal volume 104 and a fixable lid 106. The fixable lid 106 may be positioned above the opening of the insulated container 102 to seal the internal volume 104 of the insulated container. The fixable lid 106 may be positioned above the opening and secured by one or more resealable mounting mechanisms (not shown), such as one or more hinges, one or more toggles, one or more snaps, pressure tops, screw tops, etc. The walls of the fixable lid 106 and the insulated container 102 may be filled with one or more types of insulating materials, such as metal, ceramic, glass fiber, polystyrene, cellulose, foam, mineral wool, etc. For example, the fixable lid 106 and the walls of the insulated container 102 can be implemented as a double-wall vacuum insulation structure.

[0026] The internal volume 104 of the insulated container 102 can hold an organ, as well as elements and components necessary for organ perfusion and hypothermic preservation. For example, the internal volume 104 of the insulated container 102 can accommodate an organ container 108 (containing an organ 110), at least one pressure bag 124 connected to the organ by at least one infusion line 120, a sampling line 122 connected to the organ, and at least one cooling mechanism 126. The organ container 108 may be shaped and sized to accommodate an organ 110 immersed in a fixed volume of perfusion fluid 112a. The organ container 108 may be shaped and sized universally to accommodate multiple types of organs from one or more donors (e.g., based on size, complexity, etc.), and / or shaped and sized specifically to accommodate one type of organ from one or more donors. For example, the organ container 108 may be shaped and sized to fit a heart, liver, one or more lungs, one or more kidneys, pancreas, at least a portion of the small intestine, intestine, a portion of skin, a portion of tissue, a bioengineered graft, a xenograft, an arm, leg, hand, foot, fingers, toes, or other limbs. The volume, concentration, and type (e.g., blood, artificial solution, or a combination thereof) of the perfusion fluid 112a in the organ container 108 may be based on the type of organ 110 and / or the initial state of the organ. The organ container 108 may have a lid 114 that can be positioned above the opening of the organ container to seal the organ 110 inside the organ container. The lid 114 of the organ container 108 may include at least one inlet port 116 and at least one outlet port 118.

[0027] At least one infusion line 120 may enter the organ container 108 through at least one inlet port 116 and be connected to the organ 110 via cannula insertion in a vein and / or artery. A sampling line 122 may also be connected to the organ (e.g., via cannula insertion in a vein and / or artery) and be able to exit the organ container 108 through at least one outlet port 118. The sampling line 122 may be configured to allow a user of the system 100 to monitor the condition of the organ 110. For example, if the organ 110 is the liver, the system 100 may include a first infusion line and a second infusion line for supplying perfusion fluid to the liver. The first infusion line may be connected to the hepatic artery of the liver (cannula insertion), and the second infusion line may be connected to the portal vein of the liver (cannula insertion). The sampling line may be connected to the inferior vena cava of the liver.

[0028] At least one pressure bag 124 may be connected to at least one infusion line 120. For example, one pressure bag 124 may be connected to each of at least one infusion line 120 (only one is shown for ease of illustration). Each pressure bag 124 may store another volume of perfusion fluid 112b, which may be the same or different amount, type, and / or concentration. The other volume of perfusion fluid 112b may be the same type as or different from the perfusion fluid 112a in the organ container 108. Both the perfusion fluid 112a in the organ container 108 and the other volume of perfusion fluid 112b in at least one pressure bag 124 may be pre-oxygenated before use in the system 100. At least one pressure bag 124 is positioned above the organ container 108 in the inner volume 104 of the insulated container 102 and can be tilted at an angle (θ) with respect to the lid 114 of the organ container so as to allow another volume of perfusion fluid 112b to flow from the pressure bag to the organ 110 by gravity through the infusion line 120. The tilt angle (θ) of the at least one pressure bag 124 with respect to the lid 114 of the organ container 108 can be, for example, 1 to 90 degrees, 2 to 65 degrees, 3 to 45 degrees, 4 to 30 degrees, 5 to 20 degrees, etc. In one example, the tilt angle (θ) of the at least one pressure bag 124 with respect to the lid of the organ container may be 5 to 90 degrees. In another example, the tilt angle (θ) of the at least one pressure bag 124 with respect to the lid of the organ container may be 15 degrees.

[0029] At least one cooling mechanism 126 can maintain the organ at a hypothermic temperature for a certain period of time without requiring power during operation (the cooling mechanism 126 and / or other parts of the system 100 may need to be recharged or recooled using commercial power, but commercial power is not required during operation). At least one cooling mechanism 126 may be positioned at different locations within the inner volume 104 (e.g., evenly spaced around at least a portion of the organ container 108) to uniformly control the temperature of the organ for a certain period of time. The cooling mechanism 126 may be positioned on at least one side of the organ container 108 (e.g., the bottom, top, or side). In one example, the cooling mechanism 126 may be a “power-free” cooling mechanism including, for example, a gel cooling pack, a chemical coolant, and / or an ice pack. For example, the cooling mechanism 126 may be based on a chemical coolant (e.g., sodium polyacrylate), such as a eutectic gel pack. In another example, the cooling mechanism 126 may include a long-lasting, self-contained, and rechargeable cooling mechanism. The cooling mechanism 126 can cool at least a portion of the inner volume 104 of the insulated container 102 and / or the organs 110 in the organ container 108 to hypothermia and maintain that hypothermia for a certain period of time without requiring electricity. This period may be, for example, 10 hours or less, 12 hours or less, 15 hours or less, 20 hours or less, 24 hours or less, 48 ​​hours or less, or longer. The hypothermia may be, for example, 0°C to 20°C, 0°C to 14°C, 0°C to 12°C, 0°C to 11°C, or 0°C to 10°C. The hypothermia can be maintained below the desired hypothermia, for example, within 10°C, within 5°C, within 1°C, within 0.5°C, within 0.1°C, etc. In one example, the desired hypothermia may be about 10°C for 24 hours or less. The desired hypothermia can be predetermined for each organ (for example, specifically for each organ and / or for a general type of organ) to optimally slow organ damage and deterioration, such as ischemia-induced injury and deterioration.

[0030] While hypothermia is maintained by the cooling mechanism 126 and the insulation of the insulated container 102, the organ 110 may be perfused for a period of time with another volume of perfusion fluid 122b from at least one pressure bag 124 through at least one infusion line 120 cannulated into at least one vein and / or artery of the organ, without requiring power. The perfusion flow rate may be controlled by gravity (e.g., by changing the tilt angle θ of the pressure bag 124), and / or by one or more flow regulators and / or valves connected to the infusion line (not shown in Figure 1, but described in more detail in Figure 2). In some other examples, the perfusion control rate may be controlled by an external pressure applied to at least one pressure bag.

[0031] Next, referring to Figure 2, a more complex version of the dynamic organ preservation system 200 is shown. The more complex dynamic organ preservation system 200 includes all the components of the dynamic organ preservation system 100 (numbered in the 200s) and may include additional components to improve hypothermic preservation of organs and the overall utilization of the system.

[0032] The flow regulator 228 described above may be connected to at least one infusion line 220 and / or sampling line 222. The flow regulator 228 connected to the infusion line 220 can regulate the rate of another volume of perfusion fluid 212b flowing from the pressure bag 224 to the organ 208, in addition to the gravity flow. The flow regulator 228 connected to the sampling line 222 can regulate the sampling rate of the perfusion fluid (from the volume 212a in which the organ is immersed, and / or from the other volume at a point after it has flowed into the organ 208). The flow regulator 228 may be automatically and / or manually adjustable. The flow regulator 228 may be any type of valve capable of controlling the flow of fluid. At least one removable mounting mechanism 230 (shown as two mounting mechanisms for ease of illustration) can secure the organ container 208 to the insulated container 202. The removable attachment mechanism 230 has a first part connected to the organ container 208 and a second part attached to the insulated container 202, and may be a hook, snap, button, Velcro, etc. The effluent container 232 is connected to the organ 210 and / or organ container 208 by a drainage tube 234 and can receive effluent from the organ through the drainage tube 234. The effluent container 232 may be positioned below the organ container 208, for example, with a cooling mechanism 226 in between. The drainage tube 234 can exit the organ container 208 through the outlet port 218 of the lid 214 or through the side or bottom of the organ container 208.

[0033] System 200 may also include one or more sensors 236 for detecting one or more biomarkers related to the state of an organ 210 and / or for detecting changes in one or more biomarkers indicating a change in the state of the organ. For example, as shown, one or more sensors 236 may be located in the sampling line 222 or in the organ container 208 (or at any position in the line). Not shown, sensors 236 and / or one or more additional sensors may be located in at least one infusion line 220, sampling line 222, effluent container 232, the inner volume section 204 of the insulated container 202, etc. For example, one or more sensors 236 may be located in the sampling line 222 for the liver, allowing for direct sampling of effluent from at least a portion of the inferior vena cava. One or more sensors 236 may detect, for example, temperature or biomarkers such as chemical analyte concentrations (e.g., glucose, lactate, etc.), venous and / or arterial blood oxygen saturation. For example, one or more sensors 236 can detect in real time flavin mononucleotide (FMN) levels, which are markers of mitochondrial complex I damage; lactate levels, which may indicate metabolic function; bile production levels, and / or bicarbonate and sodium secretion, which may indicate the viability of bile duct cells. As another example, one or more sensors 236 can detect levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and / or lactate dehydrogenase (LDH) to predict transplant success and / or organ prognosis.

[0034] Furthermore, one or more sensors 236 may be visible (e.g., through a translucent window), accessible by temporarily removing at least a portion of the fixed lid 206 of the insulated container 202, and / or wirelessly connected to an external device. One or more sensors 236 may be powerless and / or may include a rechargeable battery (so as not to require an external power source while the system is operating). The system 200 may also include one or more transport mechanisms 238 to improve the transportability of the system. One or more transport mechanisms may include one or more handles, straps, wheels, etc. The system 200 is mobile and at least partially reusable (e.g., the insulated container 202 and fixed lid 206, the removable mounting mechanism 230, the sensors 236, etc.).

[0035] Figure 3 shows a system 300 which may include a system 200 with one or more sensors 236 wirelessly connected to an external device 340. The one or more sensors 236 may include wireless transmitters that can wirelessly connect to wireless transmitters (not shown) of the external device 340 to transmit recorded biomarker data and / or changes in one or more detected biomarkers. The external device 340 may be any device including at least a controller 342 and a display 344. For example, the external device 340 may be a computer, smartphone, tablet, or dedicated device. The controller 342 of the external device 340 may include non-temporary memory and a processor (not shown) for storing and executing instructions, respectively. The controller 342 may analyze recordings from one or more sensors 236 (such as one or more sensors in a sampling line) and generate readouts of at least one organ biomarker, organ oxygen consumption, and effluent composition. The external device 340 can alert and / or notify the user of changes in biomarkers and / or the state of organs 210 with visual, audible, and / or tactile alarms. The external device 340 can display organ health data and / or biomarkers, etc., recorded by one or more sensors 236 and / or analyzed by the controller 342 on the display 344.

[0036] Figures 4–10 illustrate an example of how to set up a dynamic organ preservation system for donor livers. Figure 4 shows the inner volume of an example of an insulated container with double-wall insulation. The inner wall of the insulated container has been modified to include multiple hooks (four are shown, but any number greater than one is conceivable, e.g., a removable mounting mechanism) for holding the organ container in place. Multiple gel packs (e.g., a cooling mechanism) are placed within the inner volume of the insulated container. Four gel packs are shown lined up along the bottom and part of the sides inside the insulated container. A total of eight gel packs were used, with others added later (before the secure lid was attached).

[0037] Figure 5 shows a liver chamber containing a liver (e.g., an organ container housing an organ) partially immersed in volume or perfusion fluid and positioned on a hook within an insulated container. In this example, the hook is connected to a hole in the outer lip of the liver container. Although not visible in the figure, an effluent container is positioned between the gel pack and the liver chamber. Figure 6 shows a magnified view of the liver chamber and liver. A real-time sampling line is positioned to extend into the subhepatic inferior vena cava (IVC). A drainage tube is positioned within the organ container. And an infusion line is shown cannula-inserted into the hepatic artery and portal vein of the liver. Figure 7 is a further magnified view of the liver and liver chamber to show that the drainage tube is positioned to receive effluent from the hepatic vein of the liver. Figure 8 shows the steps for inserting and positioning the real-time sampling tube. In element A of Figure 8, the subhepatic IVC is identified and an extension tube (e.g., a sampling tube) is inserted into the subhepatic IVC. Element B of Figure 8 shows the end of the subhepatic IVC being secured to hold the sampling tool in place. Element C in Figure 8 shows an example of a sampling tool (syringe) for collecting sample perfusion fluid from the liver.

[0038] Figure 9 shows an exemplary system with a lid placed in the liver chamber and additional gel packs positioned around the sides of the inner volume of the insulated container. The lid may be translucent to facilitate visual inspection of the liver. Several ports may be included in the lid to extend infusion lines, sampling lines, and drainage tubes outside the liver chamber. Figure 10 shows two infusion lines connected to a pressure bag, each equipped with a pressure scale and flow regulator for measuring and controlling the flow rate from the pressure bag. A thermometer was placed inside the insulated container, and another thermometer (not shown) was placed inside the liver to record the temperature over time. The pressure bag is shown tilted at a certain angle (approximately 15 degrees) relative to the lid of the liver chamber to allow the perfusion fluid to flow into the liver by gravity. The lid can then be closed over all internal components to insulate the system for a certain period of time (12 hours, as shown in the experiment). Sampling lines may extend outside the insulated container, for example, to allow an external observer to check the condition of the organ without having to open the lid of the insulated container. Figure 11 shows diagrams of other non-limiting and exemplary liver chambers (it should be understood that further configurations of liver chambers may be conceptualized). Element A shows a rounded-bottom chamber of an organ container, with a drainage tube located within the body rather than the lid, and is arranged so that the effluent can be drained when the perfusion fluid reaches the level of the drainage tube opening. Elements A and B further show tube suspension systems for supporting and / or organizing infusion lines. Element B shows a flat-bottom chamber in which the sampling line can extend out from the drainage tube rather than from the outlet port on the lid (otherwise similar to element A).

[0039] Figure 12 is a graphical representation showing how the temperature of organs and organ containers remains relatively constant for at least 12 hours. The graph shows that after the insulated container (in this example, a Yeti cooler, with the line starting at 20°C) was cooled to 11°C in 20 minutes, the temperature of the insulated container was maintained within approximately 4°C of 10°C for 12 hours. The graph also shows that the temperature of the liver (with the line starting just under 10°C) was maintained at approximately 10°C for 12 hours with fluctuations of less than approximately ±1°C.

[0040] Figure 13 shows an example of a pressure bag 324 that can be used in systems 100 and / or 200 (it should be understood that the pressure bag may have a different shape and still function in systems 100 and / or 200). The pressure bag 324 can utilize gravity and / or pressure to deliver perfusion fluid to an organ. The pressure bag 324 may include an outer housing, which may be a hard shell 350 that can protect the perfusion bag from external forces, and an internal pressure bladder 352 that can apply pressure to the perfusion bag 354. For example, the hard shell 350 may function like a clamshell with an upper and lower half that can be opened and closed around the perfusion bag 354 using hinges 356 and a locking mechanism 358. When locked in place, the pressure bladder 352 can apply a constant amount of pressure to the perfusion bag 354, allowing the perfusion fluid to flow from the perfusion outlet 360 into the infusion line at a constant rate corresponding to the applied pressure. The applied pressure can be controlled and / or modified using the pressure bladder port 362. Figure 14 shows an example of an insertion port 370 for connecting the sampling line 322 to the organ 310 (it should be understood that the insertion port may be of any shape and will still function in systems 100 and / or 200). The insertion port 370 can be incorporated into an existing opening in the organ 310 and / or placed in a window created in the organ 310. The insertion port 370 may include a flap 372 that can hold the sampling line 322 in place within the organ 310.

[0041] Figures 15–18 below show various parts of the dynamic organ preservation systems 100, 200, and / or 300, which have configurations for use in organs other than the liver. Figures 15 and 16 show exemplary diagrams of each part of the dynamic organ preservation systems 100, 200, and / or 300 for use in the kidney or a pair of kidneys. Figures 17 and 18 show exemplary diagrams of cannulas for use in the heart.

[0042] Figure 15 shows a renal cisterna container 402 for use in dynamic organ preservation systems 100, 200, and / or 300. The renal cisterna container 402 in Figure 15 may be shaped and sized to hold a single kidney 406 within a renal cisterna 404 (which is also shaped and sized to fit the kidney of a subject). The kidney 406 can be cannulated in the renal artery using a renal artery cannula 408 (e.g., connected to an infusion line) to allow perfusion fluid to be introduced into the kidney. In addition, connectors and / or cannulas that connect to a sampling port 410, and connectors and / or cannulas that connect to an effluent drainage tube 412, may also be connected to the kidney 406 (e.g., a renal vein), respectively, for monitoring physiological parameters and values ​​and for removing effluent. A ureteral stent 414 is placed inside the kidney 406 to allow urine from the kidney to be delivered to a urine collection device 416.

[0043] Figure 16 shows another version of the renal cisterna container 402 for use in dynamic organ preservation systems 100, 200, and / or 300. The renal cisterna container 402 in Figure 16 may be shaped and sized to hold two kidneys 406(1) and 406(2) within a renal cisterna 404 (which is shaped and sized to fit two kidneys as well as a portion of the renal vein and renal artery). The renal vein and renal artery can be clamped distal to the connection to the kidneys. A single renal artery cannula 408 and a connector to an effluent drainage tube 412 can be used to allow the inflow and outflow of perfusion fluid to both kidneys, respectively. An effluent sampling port 410 may be located, for example, on a portion of the renal vein connection to each kidney 406(1) and 406(2) to sample each kidney 406(1) and 406(2) individually. Urine can be collected individually from each kidney in urine collection devices 416(1) and (2). In some cases, a single urine collection device 416 may be used.

[0044] Figures 17 and 18 show illustrative diagrams of a flexible double-lumen cannula 500 for use when the organ in a cryogenic dynamic organ preservation system (e.g., systems 100, 200, and / or 300) is a heart. When the organ is a heart, one of the infusion lines for supplying perfusion fluid to the heart may be connected to the heart via brachiocephalic cannula insertion or direct aortic cannula insertion. The flexible double-lumen cannula 500 may be used with an outer cannula (coronary flow cannula 502) for supplying perfusion fluid into the heart and an inner cannula 503 for draining and / or supplying perfusion fluid and / or effusion fluid from the heart. The outer cannula (coronary flow cannula 502) may be a coronary flow cannula for supplying perfusion fluid to the region of the aortic valve near the sinoatrial junction (upward as shown) so that the perfusion fluid can flow into the left coronary artery (LCA) and the right coronary artery (RCA). The internal cannula 503 may be positioned in the left ventricle and may include a subannular balloon 504 that holds the internal cannula in the left ventricle and, if necessary, functions as a backup valve. The internal cannula 503 may function as a vent using suction and / or draw perfusion and / or effusion from inside the left ventricle using the left ventricular vent 506.

[0045] Figure 18 shows the flexible double-lumen cannula 500 in more detail. Element A of Figure 18 shows a side view of the flexible double-lumen cannula 500 with its front and rear sides marked. Element B of Figure 18 shows a front view of the flexible double-lumen cannula 500, further showing the flow port 508 connected to the coronary flow cannula 502, the suction port 510 connected to the inner cannula 503, and the balloon port 512 for inflating and / or deflating the subannular balloon 504. Element C of Figure 18 shows an internal view of the flexible double-lumen cannula 500 with the flow lumen 514 (of the coronary flow cannula 502) surrounding the suction lumen 516 (of the inner cannula 503). It should be noted that any shape, size, and / or position of the components of the flexible double-lumen cannula are shown for illustrative purposes only and may have other shapes, sizes, and / or positions known to those skilled in the art.

[0046] Figures 19–21 illustrate different exemplary cannula designs for use in any combination of the systems and devices described above. These examples are not intended to be limiting. Figure 19 illustrates an exemplary drainage cannula design (e.g., for connecting a drainage tube 234 to an organ container 208) for use in the cryodynamic organ preservation systems described above (e.g., 100, 200, 300, etc.). The drainage cannula may have a perforated tip designed to optimize drainage of the perfusion fluid, which can be positioned upward or downward depending on the embodiment of the system. The drainage cannula may include one or more grooves surrounding at least a portion of the body to facilitate fixation of the cannula into a blood vessel. For example, one or more grooves may be located adjacent to the perforated tip and be recessed to a sufficient extent to receive a knot or the like without slipping. The drainage cannula may have a tapered end with one or more ridges at the end opposite the perforated tip to ensure secure connection of the drainage cannula to a tube connected to the effluent container. Figure 20 shows a different diagram of an exemplary infusion cannula design with an elongated tip extending from a tapered body. The elongated tip can have a variety of tip lengths and widths to accommodate various blood vessel and / or organ sizes. Similar to the drainage cannula design in Figure 19, the infusion cannula design may also include a tapered other end with one or more grooves to facilitate securing the cannula into a blood vessel and / or organ using a tie, and one or more ridges to securely connect the cannula to an infusion line. Figure 21 shows an example of a vascular cannula design similar to the design in Figure 20, including a Luer connector (shown extending from the widest part of the cannula body, although it should be understood that the Luer connector may extend from any part of the body) and a tapered connector to facilitate connection to a cryodynamic organ preservation system and / or mechanical perfusion without requiring cannula changes.

[0047] method Another aspect of this disclosure may include a method 600 (Figure 22) for improving the preservation of extracorporeal organs (e.g., for transplantation). Method 600 can be carried out using systems 100, 200, or 300 shown in Figures 1, 2, and 3. For brevity, method 600 is illustrated and described as being carried out in series, but it should be understood and recognized that this disclosure is not limited by the illustrated order, as some steps may occur in a different order and / or simultaneously with other steps illustrated and described herein. Furthermore, not all illustrated embodiments are required to implement method 600, and method 600 is not limited to the illustrated embodiments.

[0048] Figure 22 illustrates a method 600 for preserving an organ outside the body using a cryogenic dynamic organ preservation system for up to 10, 12, 15, or 24 hours, or longer, without requiring external power. The dynamic organ preservation system can increase the time an organ can be safely and effectively preserved by maintaining an optimal preservation temperature and perfusing the organ. The dynamic cryogenic preservation system can uniformly cool the organ to a desired hypothermia and maintain that temperature for at least 12 or 24 hours, while simultaneously perfusing the organ using a pressure and gravity supply system. In some examples, a sensor with a reusable internal battery can be connected to an external device to monitor the organ's condition.

[0049] In 602, an organ (e.g., an organ from a donor for transplantation and / or scientific research, etc.) can be placed in an organ container with a certain volume of perfusion fluid. The organ container may be shaped and sized to fit one or more types of organs (e.g., general shapes and sizes for small, medium, or large organs), and / or may be shaped and sized specifically for a particular type of organ (e.g., heart, liver, lung, kidney, etc.). The volume and contents of the perfusion fluid placed in the organ container with the organ may be based on the specific needs of the organ. The organ may be at least partially immersed in a certain volume of perfusion fluid. In 604, at least one infusion line and a sampling line can be connected to the organ (e.g., by cannula insertion) (e.g., to at least one vein and / or artery of the organ). For example, if the organ is a liver, at least one infusion line may be a first infusion line connected to the hepatic artery of the liver and a second infusion line connected to the portal vein of the liver. A sampling line may be connected to the inferior vena cava of the liver.

[0050] In 606, the organ container can be secured within the container by closing the lid. The lid may include at least one inlet port and at least one outlet port. At least one infusion line may be passed through the inlet port and a sampling line through the outlet port of the lid. A drainage tube for the perfusion effluent may also be passed through the outlet port of the lid. The drainage tube may be connected to the organ in the “downstream” portion of the circulation and / or be in fluid communication with the volume of perfusion fluid within the organ container to remove contaminants and / or waste. In 608, the organ container may be placed within an insulated container having at least one cooling mechanism (e.g., within the inner volume of the insulated container). The organ container may be secured in place within the insulated container using one or more removable attachment mechanisms (e.g., one or more hooks, snaps, buttons, Velcro, etc., having a first part connected to the organ container and a second part attached to the insulated container). The organ container may be placed above an effluent container (which may be connected to the organ and / or organ container via a drainage tube). The cooling mechanism may be located on at least one side of the organ container (e.g., the bottom, top, or side). The cooling mechanism may be a “power-free” cooling mechanism, including, for example, a gel cooling pack, a chemical coolant, and / or an ice pack. In another example, the cooling mechanism may include a long-lasting, self-contained, and rechargeable cooling mechanism.

[0051] In 610, at least one infusion line can be connected to at least one pressure bag containing another volume of perfusion fluid (which may be the same as or different from the perfusion fluid in the organ container or other pressure bags used). The other volume of perfusion fluid may be pre-oxygenated. In 612, at least one pressure bag can be positioned above the organ container in the inner volume of the insulated container and tilted at an angle (θ) with respect to the lid of the organ container to allow another volume of perfusion fluid to flow from the pressure bag to the organ by gravity. The angle (θ) with respect to the lid of the organ container may be, for example, 5 to 90 degrees, 5 to 80 degrees, 5 to 70 degrees, 5 to 60 degrees, 5 to 50 degrees, 5 to 40 degrees, 5 to 30 degrees, etc. In one example, the angle (θ) with respect to the lid of the organ container may be 15 degrees. To control and regulate the flow of a separate volume of perfusion fluid to the organs in addition to gravity flow, a flow regulator and / or valve can be coupled to at least one infusion line (e.g., along the length of the infusion line, at the inlet port, or at a similar location). After all the components and elements described above have been positioned, secured, and / or activated, the fixable lid of the insulated container can be secured to initiate dynamic hypothermia preservation of the organs.

[0052] Dynamic hypothermia preservation of organs may include the following additional steps: In 614, the internal volume of the insulated container and the organs in the organ container can be cooled to hypothermia by at least one cooling mechanism and maintained at that hypothermia for a certain period of time without requiring power. This period may be, for example, 10 hours or less, 12 hours or less, 15 hours or less, 20 hours or less, 24 hours or less, or 48 hours or less. The hypothermia may be, for example, 0°C to 14°C, 0°C to 12°C, 0°C to 11°C, or 0°C to 10°C. The hypothermia may be maintained within, for example, 1°C, 0.5°C, or 0.1°C from the desired hypothermia. In one example, the desired hypothermia may be 10°C for 24 hours or less. The desired hypothermia may be predetermined for each organ (e.g., organ-specific and / or for general organ types) to optimally delay organ damage or deterioration such as ischemia. While hypothermia is maintained, in 616, the organ can be perfused for a certain period of time without requiring power with at least one perfusion fluid (of a different volume) from at least one pressure bag through at least one infusion line cannulated into at least one vein and / or artery of the organ. The perfusion flow rate may be controlled by gravity, external pressure applied to at least one pressure bag, and / or one or more flow regulators and / or valves connected to the infusion line. The flow regulators may be automatically and / or manually adjustable. Optionally, the concentration of oxygen and / or one or more nutrients in the perfusion fluid can be manually adjusted by briefly removing the lid from the insulated container, or automatically.

[0053] Throughout the perfusion and maintenance of hypothermia, in 618, a user (e.g., a technician, nurse, physician, or other qualified professional) can monitor the state of the organ using at least one sensor placed in the sampling line, the internal volume of the insulated container, and / or the internal volume of the organ container. If at least one sensor is placed in the sampling line, at least one sensor may be accessible when the fixable lid of the insulated container is closed (e.g., viewing through a transparent window, exiting the insulated container through a port, or similar). For example, the sampling line may allow the user to identify at least one biomarker of the organ to monitor its state. For example, based on identifying at least one biomarker, the temperature and / or perfusion flow rate may be adjusted. The at least one sensor may include a temperature sensor, an oxygen sensor, a carbon dioxide sensor, or a chemical analyte sensor (e.g., glucose, lactate, etc.). In another example, at least one of the sensors may include a rechargeable battery (so as not requiring external power) and a wireless transmitter for connecting to one or more external devices, including a display and processor / controller (e.g., a computer, tablet, smartphone, or dedicated external device). At least one sensor may wirelessly transmit collected data and / or a warning if the data at any given time exceeds a preset threshold to an external device. The external device may, for example, provide alerts to the user via sound, sight, and / or touch. The external device may optionally be in communication with a temperature control mechanism, one or more flow regulators, etc., to control the system's temperature and / or perfusion. During hypothermia preservation, the entire dynamic preservation system may be transported from one location to another (e.g., via wheels and / or handles). Transport may include carrying the entire dynamic preservation system by plane and / or car without powering the system and without using special vehicles (e.g., due to size and / or weight).

[0054] experiment The following experiment describes liver preservation using a dynamic organ preservation system (DOSS). This system maintains the liver temperature at 10°C for 10 hours while supplying oxygenated perfusion fluid in a portable cooler, allowing for real-time assessment of metabolic function. DOSS was compared to conventional static refrigeration (SCS) and to improve upon the limitations of hypothermic oxygenated mechanical perfusion (MP) (HOPE). The experiment demonstrates that livers preserved using DOSS exhibited superior liver and biliary function compared to those preserved using conventional static refrigeration (SCS). As shown, DOSS enables improvements in lactate clearance, oxygen consumption, fibrinogen synthesis, bile production, bicarbonate and sodium secretion by biliary cells, responsiveness to vasoactive agents, and liver cell viability. Technically, the experiment demonstrates that DOSS maintains the liver temperature at 10°C for 10 hours while supplying oxygenated perfusion fluid in a portable cooler, allowing for real-time assessment of metabolic function. Furthermore, DOSS enables continuous oxygenated perfusion to liver grafts at 10°C and real-time effluent sampling within a portable cooler, suggesting that DOSS can enhance the function and evaluation of DCD allogeneic grafts during organ transport, which has direct clinical applications and could contribute to expanding the donor pool. In addition, this study aimed to minimize the need for external power, improve organ quality, and reduce the size, cost, and operational complexity of organ preservation systems. Integrating the DOSS circuit into a portable cooler facilitates transport and allows for use in smaller aircraft compared to conventional devices. Therefore, DOSS has the potential to offer numerous advantages over current HOPE systems, particularly in terms of cost and logistical feasibility.

[0055] material and method Research design Liver tissue collected from a porcine DCD model was stored for 10 hours using either DOSS or SCS (n=5 for each). After storage, ambient temperature mechanical perfusion (NMP) was performed for 4 hours to assess the physiological function of the liver and bile ducts. Liver and biliary tissue was collected for histological evaluation at three time points: after collection, after 10 hours of storage, and after 4 hours of NMP. Element (a) of Figure 23 shows a schematic diagram of the experiment.

[0056] Liver harvesting in a pig DCD model The research protocol was approved by the Institutional Animal Care and Use Committee at Vanderbilt University Medical Center. This study used Yorkshire x Landrace male pigs weighing 45–54 kg. After induction of general anesthesia, donor pigs were administered a bolus of 30,000 units of heparin, and whole blood was collected using the acute normal hemodilution method. Following hypoxemic cardiac arrest, a midline laparotomy was performed, and the liver (1.380 ± 0.182 kg) was flushed through the abdominal aorta with 3 L of cold University of Wisconsin (UW) solution (CoStorSol, Preservation Solutions, Inc., Elkhorn, WI). The liver was then collected using a standard method and placed on a back table.

[0057] Back Table Procedure Element (b) of Figure 23 shows a liver graft placed in the liver chamber with cannula insertion. The liver was flushed with 1 L of UW solution via the portal vein (PV). In the DOSS group, 18G cannulas (Covidien, Medtronic, Ireland) were inserted into the hepatic artery (HA) and PV for perfusion during storage. In addition, a small sample tube was inserted into the inferior vena cava (IVC) to collect real-time effluent samples for metabolic assessment during DOSS. After 10 hours of DOSS, the cannulas and sample tubes were removed, and 12Fr, 28Fr, 36Fr, and 14Fr cannulas (Medtronic, Minneapolis, MN) were inserted into the HA, PV, IVC, and bile duct, respectively, for preparation of NMP. In the SCS group, cannulas for NMP were inserted into the blood vessels after harvesting, and the liver was stored on ice in a cooler.

[0058] Dynamic organ preservation system DOSS (see elements c and d in Figure 23) was developed to supply oxygenated perfusion fluid at 10°C within a portable cooler (YETI Coolers, LLC. Austin, TX) with a minimum power requirement of 12.96 watt-hours. A eutectic gel pack (Traferox Technologies, Inc. Mississauga, ON) was used to maintain the liver temperature at 10°C. The liver temperature was continuously monitored by a temperature probe (Pendo TECH) placed at the end of the left lobe. The liver was perfused with STEEN solution, which is used in both cell-free and cell-containing MP experiments. Prior to DOSS, the perfusion fluid was oxygenated with 2 L / min of O2 using an oxygen supply (Capiox FX15, Terumo Cardiovascular, Ann Arbor, MI) to reach a final oxygen partial pressure (pO2) of 650 mmHg or higher. In the first three experiments, porcine whole blood was added to the STEEN solution at a dilution ratio of 1:4 as an oxygen carrier. However, due to the reduced oxygen demand of liver grafts at 10°C and the improved clinical applicability of cell-free solutions, cell-free STEEN solution was used in the last two experiments. Target perfusion flow rates for HA and PV were set at 50 mL / hr and 150 mL / hr, respectively, and achieved using an infusion pump (VET / IV 2.2 Infusion Pump, Heska, Loveland, CO). The perfusion fluid was drained from the liver without recirculation and collected in a waste bag attached to the bottom of the liver chamber.

[0059] Perfusion fluid samples from the perfusion fluid bag and liver effluent samples from the IVC were evaluated every two hours using a blood gas analyzer (Element POC, Heska, Loveland, CO). Liver oxygen consumption during DOSS was calculated as described above. Levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH) in the effluent were measured at the start of DOSS and 10 hours after DOSS. FMN was measured from liver effluent samples separated from blood components by centrifugation using a Synergy HTX microplate reader (Agilent BioTek Instruments, Santa Clara, CA).

[0060] Normothermic mechanical perfusion (NMP) After 10 hours of storage, NMP was continued for 4 hours as previously reported. Briefly, the NMP system consisted of a centrifugal pump, oxygen supply, organ chamber, perfusion fluid reservoir, dialysis filter, heat exchanger, flow and saturation probes, and pressure transducer. The circuit was primed with porcine whole blood. Perfusion was initiated by gradually increasing the pump speed, with target flow rates set to 1.0–1.2 L / min for PV and 0.2–0.5 L / min for HA, respectively.

[0061] Parameters of hepatocyte function in NMP During the non-invasive multiply venous pulmonary (NMP), perfusion fluid samples from the intravascular coagulation (IVC) were collected hourly and analyzed using a blood gas analyzer. A comprehensive metabolic panel (Antech Diagnostics, Seville, OH) was measured at the start of the NMP and 4 hours after the NMP began. Fibrinogen levels were measured and evaluated as a marker of synthetic liver function. Perfusion fluid samples from the perivascular coagulation (PV) were also collected to calculate oxygen consumption. After 4 hours of NMP, 4 mg of phenylephrine was administered via HA cannula to assess the vascular reactivity of the graft to selective α1-adrenergic receptor activation.

[0062] Parameters of bile duct cell function in NMP During NMP (Natural Multiplier), bile samples were collected hourly under mineral oil for analysis of bile volume and biomarkers using a blood gas reader. Furthermore, the bile / perfusion glucose and sodium ratios were calculated to account for variations in perfusion concentration.

[0063] Histological evaluation All liver and biliary tract biopsy specimens were fixed in 10% neutral buffered formalin, embedded in paraffin, and cut into 5 μm thick sections. Hematoxylin and eosin (H&E) and trichrome staining were performed on liver tissue to assess the composite acute injury score. Apoptosis in liver tissue was evaluated by terminal deoxyuridine triphosphate nick-end labeling (TUNEL) via deoxynucleotidyltransferase and immunohistochemistry (IHC) staining for cleavage caspase-3. Biliary tract tissue sections were stained with H&E and analyzed for histological scores indicating IC (ischemic biliary injury). Immunofluorescence (IF) staining with CD31, CK7, and SOX9 was performed to quantify the perivascular plexus, cholangiocarcinoma, and progenitor cells, respectively.

[0064] Histological composite injury scores for both liver and bile duct tissues were assessed by blinded pathologists. Quantitative analysis of TUNEL, truncated caspase-3, CD31, CK7, and SOX9-positive cells was performed automatically using NIS-Elements General Analysis 3. Positive counts were defined as 0.252 mm of tissue. 2 It was standardized around the high-magnification field of view (hpf) as defined by [the specified value].

[0065] Metabolite analysis Metabolite analysis was performed on liver and biliary tract tissues collected after 4 hours of NMP using MetaboAnalyst version 5.0 (details are described in the supplementary methods). Differentially expressed metabolites were identified by volcano plots with a magnification threshold of 1.5 and p<0.05. Major metabolic pathways were identified based on pathway enrichment analysis with p<0.05 and pathway topological importance with an impact score threshold of 0.1.

[0066] statistical analysis Continuous variables are presented as mean ± standard deviation. Two-sided Student's t-tests were used to compare liver and biliary biomarkers at 4 hours between the two groups, as well as other continuous variables, including histological injury scores. A linear mixed-effects model was used to compare group responses to phenylephrine administration over time. All statistical analyses were performed using EZR, and p-values ​​less than 0.05 were considered statistically significant.

[0067] result Evaluation of liver graft status during DOSS Liver temperature remained stable throughout the 10 hours of DOSS, at 9.5±1.2°C at the start and 10.4±1.2°C at 10 hours (see element (a) in Figure 24). pO2 in cellular and cell-free perfusion fluids was 729.3±26.8 mmHg and 709.3±29.3 mmHg, respectively, at the start of DOSS, and 394.5±0.8 mmHg and 548.1±62.5 mmHg at 10 hours of DOSS (see element (b) in Figure 24). Oxygen consumption remained stable during the 10 hours of storage, measured at 0.039±0.009 mL / min / kg at 2 hours and 0.036±0.005 mL / min / kg at 10 hours (see element (c) in Figure 24). The lactate concentration in the effluent was 1.6±0.7 mmol / L at 2 hours of DOSS and 2.3±1.7 mmol / L at 10 hours (see element (d) in Figure 24). While the levels of AST, ALT, and LDH decreased numerically from (AST: 1164±743 U / L, ALT: 50±30 U / L, LDH: 1152±745 U / L) at the start of DOSS to (AST: 461±461 U / L, ALT: 27±13 U / L, LDH: 543±369 U / L) at 10 hours of DOSS, the FMN concentration increased from 150±55 ng / mL to 192±61 ng / mL (see elements (e)-(h) in Figure 24).

[0068] Hemodynamics of liver grafts during NMP and after phenylephrine administration Both DOSS and SCS livers reached target physiological levels of HA and PV flow one hour after NMP (see element (a) in Figure 25). There were no significant differences in HA and PV vascular resistance at four hours after NMP. However, after phenylephrine administration, the DOSS liver showed lower HA vascular resistance over time compared to the SCS liver (p=0.018) (see element (b) in Figure 25).

[0069] Parameters of hepatocyte function in NMP Lactate concentration in the NMP perfusion fluid decreased in the DOSS group over 4 hours of NMP (see element (a) in Figure 26). In contrast, lactate concentration increased in the SCS group from 2 to 4 hours of NMP. At 4 hours of NMP, the DOSS group showed significantly lower lactate concentrations compared to the SCS group (DOSS: 0.78 ± 0.57 mmol / L, SCS: 2.19 ± 0.83 mmol / L, p=0.023). There were no significant differences between the two groups in perfusion fluid pH, bicarbonate, and glucose (see elements (b)-(d) in Figure 26). Oxygen consumption was significantly higher in the DOSS group than in the SCS group at 4 hours of NMP (DOSS: 17.37 ± 5.99 mL / min / kg, SCS: 6.29 ± 4.43 mL / min / kg, p=0.018) (see element (e) in Figure 26). There were no significant differences in AST or ALT levels (see elements (f) and (g) in Figure 26). Fibrinogen concentration increased from 0 to 4 hours of perfusion in all livers in the DOSS group, but increased in only one liver in the SCS group. The relative increase in fibrinogen concentration from 0 to 4 hours of NMP was significantly greater in DOSS livers compared to SCS livers (DOSS: 31±8%, SCS: -35±31%, p=0.005) (see elements (h)-(i) in Figure 26).

[0070] Parameters of bile duct cell function in NMP All DOSS and SCS livers produced bile during the 4-hour NMP. However, one liver in the SCS group showed biliary bleeding and was excluded from the analysis of biliary biomarkers to mitigate the potential impact of blood contamination on the interpretation of biliary function. DOSS livers produced significantly more bile than SCS livers at 4 hours of NMP (DOSS: 11.5±3.4 mL / hr, SCS: 2.9±3.4 mL / hr, p=0.013) (see element (a) in Figure 27). Bile pH was similar between the groups at 4 hours of NMP (see element (b) in Figure 27). Bile bicarbonate was significantly higher in DOSS livers than in SCS livers (DOSS: 18.6±3.5 mmol / L, SCS: 10.2±4.0 mmol / L, p=0.035) (see element (c) in Figure 27). There were no significant differences between the groups in bile glucose concentration or bile / perfusion glucose ratio (see elements (d) and (e) in Figure 27). The bile / perfusion sodium ratio was significantly higher in DOSS livers than in SCS livers at 4 hours of NMP (DOSS: 1.05±0.02, SCS: 0.97±0.01, p=0.002) (see element (f) in Figure 27).

[0071] Histological evaluation Qualitative images of liver tissue using H&E, trichrome, cleaved caspase-3, and TUNEL are shown in element (a) of Figure 28. Histological liver injury scoring showed no statistically significant difference between the two groups at any time, including post-collection, post-storage, or post-NMP (see element (b) of Figure 28). In apoptosis assessment of liver tissue, there were no significant differences between the two groups in the number of TUNEL and cleaved caspase-3 positive cells after collection. However, after 4 hours of NMP, DOSS liver had significantly fewer TUNEL-positive cells (DOSS: 84±74 cells / hpf, SCS: 234±46 cells / hpf, p=0.009) and cleaved caspase-3 positive cells (DOSS: 127±56 cells / hpf, SCS: 263±95 cells / hpf, p=0.039) compared to SCS liver (see elements (c) and (d) of Figure 28).

[0072] Qualitative images of biliary tissue by H&E and IF are shown in element (a) of Figure 29. Biliary injury scoring did not show statistically significant differences after collection, storage, or NMP (see element (b) of Figure 29). Furthermore, there were no significant differences in the number of CD31, CK7, and SOX9-positive cells after NMP (see elements (c) and (d) of Figure 29).

[0073] From the above description, a person skilled in the art will recognize improvements, changes, and modifications. Such improvements, changes, and modifications are within the scope of the skills of a person skilled in the art and are intended to be covered by the attached claims.

Claims

1. An organ preservation system, Insulated container and The insulated container comprises a fixable lid for sealing the inner volume, The inner volume of the insulated container houses an organ container, at least one pressure bag, at least one infusion line, a sampling line, and at least one cooling mechanism. The organ container is configured to contain an organ at least partially immersed in a certain volume of perfusion fluid, the organ container has a lid configured to seal the organ container, the lid has at least one inlet port and at least one outlet port, The at least one pressure bag stores the perfusion fluid in a different volume, is positioned above the organ container, and is inclined at a predetermined angle with respect to the lid of the organ container. The at least one infusion line enters the organ container through the at least one inlet port, and connects the at least one pressure bag to the organ, allowing the perfusion fluid to flow from the at least one pressure bag into the organ by gravity. The sampling line is connected to the organ and configured to allow the user to monitor the state of the organ, and the sampling line exits the organ container through the at least one exit port. An organ preservation system characterized in that the at least one cooling mechanism is configured to maintain the organ at a hypothermic state for a certain period of time without requiring electricity.

2. The insulated container further comprises an outflow fluid container housed within the inner volume portion of the insulated container and positioned below the organ container, The organ preservation system according to claim 1, characterized in that the spillage container is a drainage tube connected to the organ and is configured to receive spillage from the organ through a drainage tube extending from the organ container to the spillage container via the at least one outlet port.

3. The organ preservation system according to claim 1, wherein the at least one infusion line and the sampling line each include a flow regulator configured to adjust the rate at which the other volume of perfusion fluid flows to the organ and a flow regulator configured to adjust the sampling rate of the perfusion fluid from the organ.

4. The organ preservation system according to claim 1, characterized in that the organ container is fixed inside the insulated container via at least one removable attachment mechanism.

5. The organ preservation system according to claim 4, characterized in that the removable attachment mechanism comprises at least one of a hook, a snap, a button, and Velcro.

6. The organ preservation system according to claim 1, wherein the predetermined angle of the organ container with respect to the lid is between 5 and 90 degrees so as to facilitate the gravity flow of the perfusion fluid of the other volume in the at least one pressure bag to the organ.

7. The organ preservation system according to claim 1, characterized in that the at least one cooling mechanism is arranged around the organ container so that the temperature of the organ is uniformly controlled for the specified period of time.

8. The organ preservation system according to claim 1, characterized in that the system is portable and at least partially reusable.

9. The organ preservation system according to claim 1, characterized in that the hypothermia is between 0°C and 14°C, and the constant time is 24 hours or less.

10. The organ preservation system according to claim 1, characterized in that the perfusion fluid is oxygenated before use of the system.

11. The organ preservation system according to claim 1, characterized in that the organ is the liver, lungs, or heart.

12. The organ preservation system according to claim 1, wherein, when the organ is the liver, the at least one infusion line is a first infusion line connected to the hepatic artery of the liver and a second infusion line connected to the portal vein of the liver, and the first infusion line and the second infusion line cause the perfusion fluid to flow into the liver.

13. The organ preservation system according to claim 12, characterized in that the sampling line is connected to the inferior vena cava of the liver.

14. The organ preservation system according to claim 1, wherein the sampling line further comprises one or more sensors configured to record and / or detect changes in one or more biomarkers indicating a change in the state of the organ.

15. With additional external devices, The aforementioned external device is The display and The organ preservation system according to claim 14, further comprising: a controller configured to analyze recordings from one or more sensors in the sampling line and generate readouts of at least one of the biomarkers of the organ, the oxygen consumption of the organ, and the composition of the effluent.

16. The organ preservation system according to claim 14, characterized in that the external device is configured to display organ health data and / or biomarkers recorded by the one or more sensors and / or analyzed by the controller.

17. A method for preserving organs at low body temperature, The steps include: placing the organ in an organ container such that the organ is at least partially immersed in a certain volume of perfusion fluid; The steps include connecting at least one infusion line and a sampling line to the organ, The step of sealing the organ container with a lid having at least one inlet port for receiving at least one infusion line and at least one outlet port for receiving the sampling line, The steps include placing the organ container in an insulated container having an inner volume, a fixable lid for sealing the inner volume, and at least one cooling mechanism disposed within the inner volume; The steps include connecting the at least one infusion line to at least one pressure bag, The steps include: positioning at least one pressure bag for storing another volume of perfusion fluid above the organ container in the insulated container, at a predetermined angle relative to the lid of the organ container, so that the other volume of perfusion fluid flows to the organ by gravity; The steps include: cooling the inner volume of the insulated container with at least one cooling mechanism within the inner volume of the insulated container, thereby maintaining the organs at a hypothermic state for a certain period of time without requiring electricity; The steps of perfusing the organ for a certain period of time without requiring electricity, through the at least one infusion line inserted via a cannula into at least one vein and / or artery of the organ using the at least one pressure bag, A method for preserving a hypothermic organ, comprising the step of enabling a user to monitor the condition of the organ using at least one sensor in the sampling line while the fixable lid is closed.

18. The method for preserving a hypothermic organ according to claim 16, further comprising the step of confirming at least one biomarker of the organ by the sampling line during the cooling and perfusion process.

19. The method for preserving a hypothermic organ according to claim 17, further comprising the step of adjusting the flow of the other volume of perfusion fluid into the organ by adjusting a flow regulator connected to at least one of the infusion lines.

20. The hypothermic organ preservation method according to claim 16, further comprising the step of moving the organ preservation system from a first location to a second location.

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