A long distance transfer and preservation device for porcine ex vivo tissue organs

Abstract

The present application relates to the technical field of medical equipment, and especially relates to a long-distance transfer and storage device for pig isolated tissue organs, which comprises a storage box, an organ storage tank, a cover, and a peristaltic pump for perfusion of the tissue organs in the organ storage tank. The present application is multi-purpose and has strong adaptability: by replacing phase change agents with different phase change temperatures in the organ storage tank jacket, the same device can meet the requirements of 35 DEG C storage of pig isolated ovaries and 4 DEG C storage of pig isolated internal organs, thereby greatly reducing the equipment cost and use threshold. The passive temperature control of the phase change agent cooperates with the active temperature control of the side wall refrigeration mechanism and the bottom heating mechanism, and cooperates with the closed-loop control of the temperature sensor and the controller, so that high uniformity and high stability of the temperature in the organ storage tank are realized. The partition plate, the hole plate, the suspension storage liquid and the damping pad block constitute a multiple mechanical protection system, which effectively prevents the mechanical damage of the tissue organs during long-distance transportation.

Description

Technical Field

[0001] This invention relates to the field of medical device technology, and in particular to a long-distance transfer and preservation device for ex vivo porcine tissues and organs. Background Technology

[0002] Organ transplantation is the most effective treatment for end-stage organ failure. Taking kidney transplantation as an example, the donor organ is often not located in the same place as the recipient, requiring long-distance transport of the organ to the recipient's location for surgery. Maintaining the viability of the organ during preservation and transportation is a crucial prerequisite for successful organ transplantation.

[0003] In the fields of in vitro embryo production and research in livestock, as well as organ transplantation, the long-distance transfer and preservation of isolated porcine tissues and organs is a key technological bottleneck restricting the development of related research. Porcine ovaries contain follicles of different developmental types and are important raw materials for embryo engineering technologies such as in vitro oocyte maturation, fertilization and development, and somatic cell cloning. Slaughterhouses, which provide porcine ovaries, are often located far from laboratories, requiring temperature-controlled preservation and transportation to ensure the viability of isolated ovarian tissue cells. However, porcine oocytes are highly sensitive to temperature stress; temperatures below 16°C affect cell membrane structure, while 30–35°C is the optimal temperature for preservation and transportation, providing the best conditions for porcine oocyte maturation and development.

[0004] Furthermore, cryopreservation is a crucial step in the transplantation research of excised porcine internal organs (such as the heart, liver, and kidneys). Excised porcine organs typically require perfusion and preservation at around 4°C to reduce cellular metabolic rate and minimize ischemic damage.

[0005] However, existing technologies lack a universal long-distance transfer and preservation device capable of simultaneously meeting the requirements of the two different preservation temperatures mentioned above. Existing devices are typically only suitable for the preservation of a single type of tissue or organ under a single temperature condition, with limited functionality and poor adaptability. Furthermore, existing devices are inadequate in terms of the sealing of the organ preservation tank, temperature uniformity control, and vibration damping protection, making it difficult to meet the complex requirements of long-distance transportation of ex vivo porcine tissues and organs. Summary of the Invention

[0006] The purpose of this invention is to provide a long-distance transfer and preservation device for isolated pig tissues and organs. By changing the phase change agent, it can flexibly adapt to the preservation temperature requirements of different tissues and organs, and has good sealing performance, temperature uniformity and vibration reduction protection functions. It is suitable for long-distance transfer and preservation of isolated pig ovaries (preserved at around 35°C) and isolated pig internal organs (preserved at around 4°C).

[0007] To solve the above-mentioned technical problems, the present invention adopts the following technical solution: This invention provides a long-distance transfer and preservation device for isolated porcine tissues and organs, comprising a preservation box, an organ preservation tank disposed within the preservation box, a cover covering the preservation box, and a peristaltic pump for perfusing the tissues and organs in the organ preservation tank; The storage box is divided into two cavities by an insulating partition, which are used to install the organ storage tank and to house the peristaltic pump, respectively. The organ preservation tank is a jacketed structure, and a phase change agent is filled inside the jacket. A side wall cooling mechanism and a bottom heating mechanism are respectively provided on the outer wall and bottom of the organ preservation tank. Multiple horizontally and vertically intersecting partitions can also be detachably installed inside the organ preservation slot to form a grid-like storage slot for individually preserving animal organs.

[0008] Furthermore, a side wall heat dissipation groove is provided on the storage box and on the side wall corresponding to the side wall cooling mechanism; a heat insulation layer is provided at the bottom of the storage box and at the bottom corresponding to the bottom heating mechanism.

[0009] Furthermore, the cover is attached to the storage box by a snap or buckle; Wherein, a first sealing ring is provided on the cover corresponding to the upper end face of the storage box; a second sealing ring is provided on the cover and at a position corresponding to the upper end face of the organ storage slot; The second sealing ring is an inflatable and deformable sealing airbag; A pre-reserved arc-shaped groove adapted to the second sealing ring is provided around the upper end face of the organ preservation slot; an inflation valve communicating with the second sealing ring is provided on the top surface of the cover.

[0010] Furthermore, a preservation solution is injected into the organ preservation tank, the density of which is close to that of the tissue or organ, so that the tissue or organ is suspended in the preservation solution.

[0011] Furthermore, a vibration damping pad is provided at the bottom of the storage box to support the bottom and side walls of the organ storage tank.

[0012] Furthermore, a perforated plate is provided in the organ preservation tank, and through holes are evenly spaced on the perforated plate. The perforated plate is made of a flexible material to prevent the tissues and organs from directly contacting the bottom plate of the organ preservation tank.

[0013] Furthermore, a temperature sensor is installed inside the organ preservation tank, and the temperature sensor is electrically connected to a controller that controls the side wall cooling mechanism and the bottom heating mechanism respectively.

[0014] Furthermore, an infusion cooling coil and an outlet conduit are also provided in the organ preservation tank. One end of the infusion cooling coil extending outside the organ preservation tank is connected to the peristaltic pump, and the other end is connected to the inlet blood vessel of the organ. One end of the outlet conduit extending outside the organ preservation tank is connected to a waste liquid storage tank, and the other end is connected to the outlet blood vessel of the organ. A flow rate sensor is provided on the outlet conduit, and the flow rate sensor and the peristaltic pump are electrically connected to a controller.

[0015] Furthermore, the inlet pipe of the peristaltic pump is connected to the perfusion bag, and it is also connected to the perfusion cooling coil through a supply pipe, for cooling the perfusion agent in the perfusion bag before it enters the tissue or organ; the perfusion agent is a hypertonic citrate purine solution.

[0016] Furthermore, a flow rate sensor is also installed on the liquid supply pipe, and the flow rate sensor is electrically connected to a controller, which in turn controls an alarm.

[0017] Compared with the prior art, the beneficial technical effects of the present invention are as follows: (1) Multi-purpose and highly adaptable: By changing the phase change agent with different phase change temperature in the organ preservation tank jacket, the same set of equipment can meet the two different temperature requirements of 35°C preservation of pig ovaries and 4°C preservation of pig internal organs, which greatly reduces equipment cost and usage threshold.

[0018] (2) Precise temperature control and good stability: The phase change agent passive temperature control works in conjunction with the side wall cooling mechanism and the bottom heating mechanism active temperature control, and with the closed-loop control of the temperature sensor and controller, the temperature in the organ preservation tank is highly uniform and stable.

[0019] (3) Multiple protections and high safety: The partition plate, perforated plate, suspension preservation fluid and vibration damping pad constitute a multiple mechanical protection system, which effectively prevents tissues and organs from being mechanically damaged during long-distance transportation.

[0020] (4) Reliable sealing and pollution prevention: The first sealing ring and the second sealing ring (inflatable sealing airbag) form a double sealing structure, which effectively prevents leakage of the storage liquid and the entry of external pollutants.

[0021] (5) Complete perfusion function: For visceral organs that require continuous perfusion, the peristaltic pump, perfusion cooling coil, flow rate sensor and controller constitute a complete perfusion circuit to realize low temperature perfusion preservation and extend the in vitro preservation time of organs.

[0022] (6) Easy to operate and highly practical: The partition is detachable and can be flexibly adjusted according to actual needs; the cover is fixed by buckle or snap fastener, making it easy to open and close; the inflation valve is simple to operate and has a reliable seal. Attached Figure Description

[0023] The present invention will be further described below with reference to the accompanying drawings. Figure 1 This is a schematic diagram of the structure of the long-distance transfer and preservation device for isolated porcine tissues and organs according to the present invention; Figure 2 A schematic diagram of the structure of a pig ovarian organ preservation tank; Figure 3 A three-dimensional schematic diagram of the structure of a pig ovarian organ preservation tank; Figure 4 This is a schematic diagram of the casing of the long-distance transfer and preservation device for isolated porcine tissues and organs according to the present invention; Figure 5 for Figure 2 Internal structure diagram; Figure 6 This is a schematic diagram of a perfusion system for isolated porcine tissues and organs.

[0024] Explanation of reference numerals in the attached drawings: 1. Storage box; 11. Insulation partition; 12. Insulation layer; 13. Side wall heat dissipation groove; 14. Cover; 141. First sealing ring; 142. Second sealing ring; 2. Organ storage tank; 21. Phase change agent; 22. Filling cooling coil; 23. Perforated plate; 231. Through hole; 24. Storage liquid; 25. Divider plate; 3. Side wall cooling mechanism; 4. Bottom heating mechanism; 5. Peristaltic pump; 51. Liquid supply pipe; 52. Filling bag; 6. Tissue / organ; 7. Vibration damping pad; 8. Liquid outlet pipe; 81. Waste liquid storage tank; 9. Flow sensor; 10. Alarm. Detailed Implementation

[0025] Example 1 This invention provides a long-distance transfer and preservation device for isolated porcine tissues and organs, comprising a preservation box 1. The preservation box 1 has a rectangular parallelepiped structure, is made of heat-insulating material, and has walls with a thickness of 10mm to 30mm. The inner and outer wall partitions are filled with heat-insulating material, with a thickness of not less than 5mm, to reduce heat exchange between the inside and outside. Rubber vibration-damping pads are installed at the four corners of the bottom of the preservation box 1 to absorb some vibration energy during transportation.

[0026] The top of the storage box 1 is equipped with a cover 14, which is detachably attached to the storage box 1 by means of a snap or a buckle. A first sealing ring 141 is installed between the cover 14 and the upper surface of the storage box 1. The first sealing ring 141 is an elastic sealing ring that is embedded in a sealing groove around the upper surface of the storage box 1 to achieve overall sealing of the storage box 1.

[0027] Inside the storage box 1, an insulating partition 11 divides the inner cavity of the storage box 1 into two independent chambers, left and right. The left chamber is used to install the organ preservation tank 2, and the right chamber is used to house the peristaltic pump 5 and the infusion bag 52, etc. The insulating partition 11 is made of polyurethane foam or other low thermal conductivity insulation material, with a thickness of 5mm to 15mm, effectively preventing the heat generated by the peristaltic pump 5 and other equipment in the right chamber from being conducted to the left chamber.

[0028] refer to Figure 1 The organ preservation slot 2 is installed in the left cavity of the preservation box 1. The organ preservation slot 2 is made of a metal material with good thermal conductivity (such as aluminum alloy or stainless steel) and has an overall box-shaped structure with an open top.

[0029] refer to Figure 2 or Figure 5 The organ preservation tank 2 has a jacket structure, meaning its sidewalls and bottom plate are both double-layered, forming a closed interlayer space between the inner and outer walls. This interlayer space is filled with a phase change agent 21. The amount of phase change agent 21 fills 70% to 90% of the interlayer space volume, leaving some space for volume expansion during phase change.

[0030] A side-wall cooling mechanism 3 is installed on the outer wall of the organ preservation tank 2, and a bottom heating mechanism 4 is installed at the bottom of the organ preservation tank 2. The side-wall cooling mechanism 3 is a semiconductor cooler that uses the Peltier effect principle for cooling; the bottom heating mechanism 4 is an electric heating mechanism. Specifically, the semiconductor cooler is selected as a TEC1-12706 or equivalent device, with its cold end in close contact with the outer wall or bottom wall of the organ preservation tank 2 and its hot end facing outward.

[0031] refer to Figure 1 On the side wall of the storage box 1, a side wall heat dissipation groove 13 is provided corresponding to the position of the side wall cooling mechanism 3. The side wall heat dissipation groove 13 has a grid-like opening, which facilitates the dissipation of heat generated at the hot end of the semiconductor cooler to the external environment. A small cooling fan can also be added at the heat dissipation groove to enhance convective heat dissipation and improve cooling efficiency.

[0032] A heat insulation layer 12 is provided at the bottom of the storage box 1 and at the bottom corresponding to the bottom heating mechanism 4. The heat insulation layer 12 is attached to the inner wall of the bottom of the storage box 1 to reduce the loss of heat at the bottom.

[0033] A temperature sensor is installed inside the organ preservation tank 2. The temperature sensor is a platinum resistance temperature sensor (Pt100) with a measurement accuracy of ±0.1℃. The signal output terminal of the temperature sensor is electrically connected to the signal input terminal of the controller, and the control signal output terminal of the controller is electrically connected to the control terminals of the side wall cooling mechanism 3 and the bottom heating mechanism 4. The controller uses a PID (proportional-integral-derivative) control algorithm to adjust the input current of the semiconductor cooler based on the deviation between the real-time temperature fed back by the temperature sensor and the target temperature, thereby achieving precise temperature control within the organ preservation tank 2.

[0034] refer to Figure 1 As shown, preservation solution 24 is injected into the organ preservation tank 2. Preservation solution 24 is a standard solution for organ preservation, and its density is adjusted to be similar to that of isolated pig tissue / organ 6 by adding appropriate solutes (such as hydroxyethyl starch, glucose, etc.). The density of a pig kidney is approximately 1.05 g / cm³. 3 Therefore, the density of the preservation solution 24 was adjusted to 1.03–1.07 g / cm³. 3 Within this range, the tissues and organs 6 are kept in a suspended or micro-suspended state in the preservation solution 24, preventing the organs from sinking directly to the bottom and being compressed.

[0035] A perforated plate 23 is installed at the bottom of the organ preservation tank 2. The perforated plate 23 is made of flexible material, such as medical-grade silicone or polyurethane foam, with a thickness of 3mm to 8mm. Through holes 231 are evenly spaced on the perforated plate 23, with a diameter of 2mm to 5mm and a spacing of 5mm to 10mm. The through holes 231 ensure free flow of the preservation solution 24 within the organ preservation tank 2, ensuring that all parts of the organ can contact the preservation solution 24. The flexible perforated plate 23 further isolates the organ from rigid contact with the bottom of the organ preservation tank 2, preventing damage caused by collisions between the organ and the tank bottom during transport.

[0036] refer to Figure 2 and Figure 3 Multiple horizontally and vertically intersecting partitions 25 are detachably installed within the organ preservation tank 2 to form a grid-like storage compartment for individually preserving animal organs. The partitions 25 are installed on the inner wall of the organ preservation tank 2 using a detachable plug-in structure, and the number and size of the grid can be adjusted according to actual needs. The partitions 25 are installed when multiple small organs (such as pig ovaries) need to be preserved, and can be removed to obtain more storage space when large organs (such as pig kidneys or hearts) need to be preserved.

[0037] A vibration-damping pad 7 is installed between the bottom of the storage box 1 and the organ storage tank 2. The vibration-damping pad 7, made of a high-damping elastic material (such as polyurethane or viscoelastic damping material) and with a thickness of 5mm to 15mm, surrounds the bottom and side walls of the organ storage tank 2. The vibration-damping pad 7 serves two purposes: firstly, it supports the organ storage tank 2, keeping it stable within the storage box 1; secondly, it absorbs and dissipates vibration energy during transportation, reducing the transmission of external vibrations to the organ storage tank 2.

[0038] refer to Figure 1 The cover 14 is detachably attached to the upper surface of the storage box 1 via a snap or buckle. The cover 14 is flat and made of heat-insulating material. A first sealing ring 141 is installed on its lower surface where it contacts the upper surface of the storage box 1. The first sealing ring 141 is an annular elastic sealing ring, embedded in a sealing groove around the lower surface of the cover 14 or around the upper surface of the storage box 1. When the cover 14 is closed, the first sealing ring 141 is compressed and deformed, filling the gap between the cover 14 and the storage box 1, achieving an overall seal of the storage box 1.

[0039] A second sealing ring 142 is installed on the lower surface of the cover 14, corresponding to a position around the upper end of the organ preservation slot 2. The second sealing ring 142 is an inflatable and deformable sealing airbag made of a flexible, airtight material (such as silicone or rubber), with an internal annular hollow structure. A pre-reserved arc-shaped groove, adapted to the shape of the second sealing ring 142, is formed around the upper end of the organ preservation slot 2. An inflation valve, a one-way valve, is provided on the top surface of the cover 14, connecting to the internal cavity of the second sealing ring 142. Gas can be injected into the second sealing ring 142 via an external air source (such as a manually inflatable airbag or a small air pump).

[0040] In use, first close the cover 14 and lock it with a snap or buckle; then inflate the second sealing ring 142 through the inflation valve. After the sealing airbag expands, it embeds into the reserved arc-shaped groove on the upper end face of the organ preservation slot 2, completely sealing the upper end of the organ preservation slot 2. When it is necessary to open, first release the gas in the second sealing ring 142 through the inflation valve. After the sealing airbag contracts, it disengages from the reserved arc-shaped groove, and the cover 14 can be opened.

[0041] In the double sealing structure, the first sealing ring 141 achieves overall sealing of the storage box 1 to prevent external contaminants from entering the storage box 1; the second sealing ring 142 achieves independent sealing of the organ preservation tank 2 to prevent the preservation liquid 24 from overflowing or leaking from the upper port of the organ preservation tank 2 during transportation.

[0042] refer to Figure 6 The injection system includes a peristaltic pump 5, an injection bag 52, a supply pipe 51, an injection cooling coil 22, an outlet pipe 8, and a waste liquid storage tank 81.

[0043] The peristaltic pump 5 is installed in the right cavity of the storage box 1, and its inlet end is connected to the perfusion bag 52 via a pipeline. The perfusion bag 52 contains the perfusion solution. The perfusion solution is a hypertonic citrate purine solution (i.e., HC-A solution), which is a standard solution used clinically in China for the perfusion and preservation of isolated kidneys. It contains effective components such as potassium citrate, sodium citrate, magnesium sulfate, mannitol, and adenine. The perfusion bag 52 is a sterile medical infusion bag, which is suspended on a hook on the inner wall of the right cavity.

[0044] The outlet end of the peristaltic pump 5 is connected to the inlet end of the perfusion cooling coil 22 via a supply pipe 51. The perfusion cooling coil 22 is installed inside the organ preservation tank 2 and is immersed in the preservation solution 24. The perfusion cooling coil 22 is made of a metal with good thermal conductivity (such as stainless steel or copper) and is in a spiral or serpentine bend shape to increase the flow rate and heat exchange area of ​​the perfusion agent within the coil. The outlet end of the perfusion cooling coil 22 is connected to the inlet blood vessel (such as the renal artery) of the tissue organ 6 via a pipeline.

[0045] When the peristaltic pump 5 is started, the hypertonic citrate purine solution in the perfusion bag 52 is pumped into the supply pipe 51. When it flows through the perfusion cooling coil 22, the perfusion agent exchanges heat with the preservation solution 24 at 4°C in the organ preservation tank 2. The perfusion agent is cooled to about 4°C and then enters the inlet blood vessel of the tissue organ 6 through the pipeline to perform low-temperature mechanical perfusion of the organ.

[0046] One end of the outlet conduit 8 is connected to the outlet blood vessel (such as the renal vein) of the organ 6, and the other end extends out of the organ preservation tank 2 and is connected to the waste liquid storage tank 81. After the infusion fluid flows through the inside of the organ 6, it is led out by the outlet conduit 8 and flows into the waste liquid storage tank 81 for collection. The waste liquid storage tank 81 is a sealed container installed in the right cavity of the preservation box 1.

[0047] In this embodiment, a first flow rate sensor 91 is installed on the outlet conduit 8. Simultaneously, a second flow rate sensor 92 is also installed on the supply conduit 51. The flow rate sensors are non-invasive ultrasonic flow sensors; the sensor body is clamped to the outer wall of the pipe, avoiding contact with the injection agent and thus preventing contamination risks. The measurement accuracy is ±2%.

[0048] The signal output terminals of the first flow rate sensor 91 and the second flow rate sensor 92 are electrically connected to the signal input terminal of the controller. The controller has a preset allowable deviation threshold (e.g., ±5%) between the inlet and outlet flow rates. During device operation, the controller receives the inlet flow rate data measured by the second flow rate sensor 92 on the supply pipe 51 and the outlet flow rate data measured by the first flow rate sensor 91 on the outlet pipe 8 in real time, and calculates the difference between the two.

[0049] When the difference between the inlet and outlet flow rates continuously exceeds a preset threshold (e.g., more than 10 seconds), the controller determines that a leak has occurred in the infusion circuit and immediately sends an alarm signal to the alarm 10. The alarm 10, installed on the outer wall of the storage tank 1, is an audible and visual alarm; upon receiving the alarm signal, it emits an audible and visual alarm to alert the operator to handle the situation promptly. Simultaneously, the controller can also send a stop signal to the peristaltic pump 5 to automatically stop the infusion process, preventing significant leakage of the infusion agent and potential losses or contamination.

[0050] Example 2 This embodiment uses the long-distance transfer and preservation of isolated porcine ovaries as an example to provide a detailed description of the device and its usage method of the present invention.

[0051] Before preserving isolated porcine ovaries, the type of phase change agent 21 was selected based on the required preservation temperature of 35°C. The original low-temperature phase change agent in the jacket was replaced with a medium-temperature phase change agent with a phase change temperature of 30–35°C. Specifically, in this embodiment, the phase change agent 21 was selected as a medium-temperature phase change agent with a phase change temperature of 30–35°C; the mixture of calcium nitrate tetrahydrate and magnesium nitrate hexahydrate was used as the phase change material, with a phase change temperature between 31 and 33°C.

[0052] This type of phase change material has a high latent heat of phase change, which can continuously release heat during long-distance transportation and maintain the temperature inside the organ preservation tank 2 at around 35°C.

[0053] The phase change agent 21 is filled to 70% to 90% of the jacket layer volume, leaving some space for volume expansion during phase change. During filling, ensure that the phase change agent 21 is evenly distributed to avoid residual air bubbles affecting heat exchange efficiency.

[0054] The bottom heating mechanism 4 is activated. During the initial stage of transportation, when the ambient temperature is below 35°C, the bottom heating mechanism 4 activates to ensure that the temperature inside the organ preservation tank 2 quickly reaches and is maintained at approximately 35°C. A semiconductor cooler (sidewall cooling mechanism 3) assists in cooling when the temperature is too high. A temperature sensor monitors the temperature inside the organ preservation tank 2 in real time, and the controller controls the activation and deactivation of the sidewall cooling mechanism 3 and the bottom heating mechanism 4 according to a preset temperature threshold (35±1°C in this embodiment), achieving precise temperature control.

[0055] The partition plate 25 is installed inside the organ preservation slot 2 to form a grid-like storage slot. Based on the size and number of pig ovaries, in this embodiment, the partition plate 25 is configured with 5 horizontal grids and 6 vertical grids, forming a total of 30 grid-like storage slots, each capable of holding one pig ovary. The partition plate 25 is installed using a detachable plug-in structure, allowing adjustment of the number and size of the grids as needed.

[0056] Preservation solution 24 is injected into the organ preservation tank 2. In this embodiment, preservation solution 24 is 0.9% physiological saline supplemented with penicillin (100 IU / mL) and streptomycin (100 mg / mL), and its density is adjusted to match that of porcine ovarian tissue (approximately 1.04–1.05 g / cm³) by adding hydroxyethyl starch. 3 The similarity allows the pig ovary to be suspended in the preservation solution 24, avoiding direct contact with the bottom of the organ preservation tank 2 and preventing mechanical damage.

[0057] The flexible perforated plate 23 is laid at the bottom of the organ preservation tank 2 to prevent the tissues and organs 6 from directly contacting the bottom plate of the organ preservation tank 2. The evenly spaced through holes 231 on the perforated plate 23 facilitate the free flow of the preservation solution 24 and the uniform temperature distribution.

[0058] The ovaries of slaughtered pigs are immediately removed, cleaned, and transported to the laboratory using this device. Oocytes are then extracted from the follicles in a clean bench. The cleaned ovaries are placed one by one into a grid-like storage compartment formed by the partition plate 25, with the cortical side of the ovary facing upwards. The partition plate 25 effectively prevents the ovaries from colliding and compressing each other, while also facilitating classification, labeling, and management. In this embodiment, 30 grid-like storage compartments can simultaneously hold 30 ovaries, meeting the needs of batch collection and transportation.

[0059] The cover 14 is closed and secured to the storage box 1 using a snap or buckle. Inflation is applied to the second sealing ring 142 (sealing airbag) via the inflation valve, causing it to expand and deform, tightly embedding into the pre-reserved arc-shaped groove on the upper surface of the organ storage tank 2, thus achieving independent sealing of the organ storage tank 2. Simultaneously, the first sealing ring 141 achieves overall sealing of the storage box 1. This double-sealing structure effectively prevents leakage of the storage solution 24 and the entry of external contaminants, while also reducing the evaporation of moisture from the storage solution 24.

[0060] The phase change agent 21 inside the jacket undergoes a solid-liquid phase change at 30–35°C, absorbing or releasing a large amount of latent heat during the phase change process. When the ambient temperature is higher than the phase change temperature, the phase change agent 21 absorbs heat through melting, inhibiting the temperature rise in the organ preservation tank 2; when the ambient temperature is lower than the phase change temperature, the phase change agent 21 releases heat through solidification, inhibiting the temperature drop.

[0061] The bottom heating mechanism 4 acts as an active compensation heating source, activating when the ambient temperature is too low to prevent the temperature from dropping below 35°C. The side wall cooling mechanism 3 activates to assist in cooling when the ambient temperature is too high. Since the semiconductor cooler has bidirectional temperature control capability (heating can be generated by passing a reverse current), in some embodiments, the side wall cooling mechanism 3 can also assist in heating.

[0062] The insulation layer 12 at the bottom of the storage box 1 effectively reduces heat loss from the bottom, while the heat dissipation grooves 13 on the side wall facilitate heat dissipation of the side wall cooling mechanism 3, ensuring cooling efficiency.

[0063] During transportation, the vibration damping pads 7 installed at the bottom of the preservation box 1 effectively absorb and buffer vibrations and impacts from the transport vehicle, protecting the organ preservation tank 2 and the internal pig ovaries from mechanical damage. The rubber vibration damping pads at the four corners of the bottom of the preservation box 1 further attenuate high-frequency vibrations.

[0064] At 35℃, pig ovaries can be preserved and transported for 2-3 hours. Upon arrival at the destination, open the cover 14, first release the gas in the second sealing ring 142 to cause the sealing airbag to contract and disengage from the reserved arc groove, then open the latch or buckle and remove the cover 14.

[0065] Remove the porcine ovaries from the separator plate 25 and each grid, and wash them three times in physiological saline containing penicillin and streptomycin. Then, use a 10mL syringe with an 18G needle to aspirate follicular fluid. Under a stereomicroscope, select cumulus-oocyte complexes (COCs). Select COCs with uniform cytoplasm and more than three layers of cumulus cells for subsequent in vitro maturation culture (IVM), in vitro fertilization (IVF), in vitro development (IVC), or cloning experiments.

[0066] Example 3 This embodiment uses the long-distance transfer and preservation of isolated porcine kidneys as an example to describe in detail the device and method of use of the present invention. The preservation methods for isolated porcine hearts, livers, and other internal organs are similar.

[0067] Before preserving pig internal organs, the type of phase change agent 21 is selected based on the required preservation temperature of 4°C. In this embodiment, a phase change material with a phase change temperature of 4°C is selected as the phase change agent 21. Specifically, tetrabutylammonium bromide (TBAB) solution or n-decyl alcohol / lauric acid binary composite solution can be used. Taking the n-decyl alcohol / lauric acid binary composite solution as an example, n-decyl alcohol and lauric acid are compounded in a molar ratio of 36:64. Its initial phase change temperature is approximately 3.80°C, with a high latent heat of phase change, stable chemical properties, and no supercooling or phase separation phenomenon. The filling amount of phase change agent 21 accounts for 70% to 90% of the space volume of the jacket layer.

[0068] The sidewall cooling mechanism 3 is activated. In the 4°C low-temperature preservation mode, the sidewall cooling mechanism 3 is the primary temperature control method. After the semiconductor cooler is powered on, the cold end is in close contact with the outer and bottom walls of the organ preservation tank 2, continuously extracting heat from the phase change agent 21 and the inside of the organ preservation tank 2; the hot end dissipates heat to the external environment through the sidewall heat dissipation grooves 13. The bottom heating mechanism 4 is activated when the ambient temperature is too low to prevent the temperature from dropping excessively below the freezing point and causing ice crystal damage.

[0069] A temperature sensor monitors the temperature inside the organ preservation tank 2 in real time. The controller controls the start and stop of the side wall cooling mechanism 3 and the bottom heating mechanism 4 according to a preset temperature threshold (4±1℃ in this embodiment). The insulation layer 12 at the bottom of the preservation tank 1 effectively isolates external heat from entering, and the heat dissipation grooves 13 on the side wall ensure the heat dissipation efficiency of the cooling mechanism.

[0070] In this embodiment, the choice of whether to install the partition plate 25 depends on the size of the internal organs. For larger single internal organs (such as kidneys or hearts), the partition plate 25 may not be installed, and the organ may be placed directly in the organ storage slot 2. For multiple small organs, the partition plate 25 may be installed to form a grid-like storage slot for classified storage.

[0071] Preservation solution 24 is injected into organ preservation tank 2. In this embodiment, preservation solution 24 is a hypertonic citrate purine solution (HC-A solution), which is pre-cooled to 1-4°C before use. The density of preservation solution 24 is adjusted to be similar to that of porcine kidney tissue (approximately 1.05 g / cm³). 3 )similar.

[0072] A flexible perforated plate 23 is laid at the bottom of the organ preservation tank 2 to prevent internal organs from directly contacting the bottom plate of the organ preservation tank 2 and causing local pressure damage.

[0073] After obtaining the isolated pig kidney, the warm ischemia time should be minimized (less than 5 minutes). The kidney should be placed in physiological saline at 4°C, and the perirenal fat and renal capsule should be quickly separated to identify the renal artery and renal vein.

[0074] Under 4°C operating conditions, the hypertonic citrate purine solution infusion bag (infusion bag 52) is connected to the inlet pipe of the peristaltic pump 5. The outlet end of the peristaltic pump 5 is connected to the inlet end of the infusion cooling coil 22 via the supply pipe 51. The outlet end of the infusion cooling coil 22 is connected to the renal artery (infusion vessel) via sterile tubing. One end of the outlet conduit 8 is connected to the renal vein (outflow vessel) via sterile tubing, and the other end is connected to the waste liquid storage tank 81.

[0075] Before perfusion, turn on the peristaltic pump 5 to perfuse at a low speed to purge air from the pipeline and the perfusion cooling coil 22, ensuring that there are no air bubbles left in the pipeline and preventing air emboli from entering the renal microvessels and causing damage.

[0076] Start the peristaltic pump 5 and perfuse the pre-cooled hypertonic citrate purine solution (pre-cooled to 1-4°C) into the kidney through the perfusion cooling coil 22 at an appropriate flow rate (e.g., for the kidney, the flow rate corresponding to a water column pressure of about 1 meter, usually 50-100 mL / min).

[0077] The perfusion cooling coil 22 is immersed in the 4°C preservation solution 24 in the organ preservation tank 2. When the perfusion agent (hypertonic citrate purine solution) flows through the perfusion cooling coil 22, it undergoes sufficient heat exchange with the 4°C preservation solution 24 and is cooled to about 4°C. Then, it enters the renal artery through the pipeline to perform low-temperature mechanical perfusion of the kidney.

[0078] During perfusion, the perfusion fluid flows through the renal arterial system → glomerular capillaries → renal vein system, and then flows into the waste fluid storage tank 81 through the outflow catheter 8. Perfusion continues until the kidneys are uniformly pale and there are no red patches on the surface, indicating that the blood in the kidneys has been fully replaced. The perfusion volume is generally 2 to 3 times the volume of the kidneys.

[0079] After perfusion is complete, peristaltic pump 5 is maintained at a low flow rate (20–50 mL / min) to continue perfusion and maintain the microcirculation of the kidney in an isolated state. Continuous hypothermic mechanical perfusion not only reduces the organ's metabolic rate but also continuously removes metabolic waste and prevents microvascular collapse and endothelial cell damage.

[0080] After perfusion, the kidney is completely immersed in a 4°C hypertonic citrate-purine solution 24 within the organ preservation tank 2. Ensure that all surfaces of the kidney are in full contact with the preservation solution 24.

[0081] The cover 14 is closed and locked with a snap or latch. Inflation is applied to the second sealing ring 142 via the inflation valve, causing the sealing airbag to expand and embed into the pre-reserved arc-shaped groove on the upper surface of the organ preservation tank 2, achieving independent sealing of the organ preservation tank 2. The first sealing ring 141 achieves overall sealing of the preservation box 1.

[0082] During transportation, the side-wall cooling mechanism 3 operates continuously (or intermittently), working in conjunction with the phase change agent 21's phase change cold storage characteristics to maintain the temperature inside the organ preservation tank 2 at approximately 4°C. The phase change agent 21 undergoes a liquid-solid phase change at 4°C, releasing or absorbing latent heat, thus suppressing temperature fluctuations within a very small range.

[0083] During the infusion process, the second flow rate sensor 92 on the supply pipe 51 and the first flow rate sensor 91 on the outlet pipe 8 monitor the inlet and outlet flow rates in real time and transmit the data to the controller. The controller calculates the difference between the two in real time. When the difference continuously exceeds a preset threshold, it determines that a leak has occurred, drives the alarm 10 to sound an alarm, and stops the peristaltic pump 5 to prevent a large leakage of the infusion agent from causing organ ischemia damage or contamination of the storage box.

[0084] The vibration damping pad 7 absorbs and buffers vibration impacts during transportation, protecting the organ preservation tank 2 and the delicate vascular structure and tissue integrity of the kidney inside.

[0085] At 4°C, isolated porcine kidneys can be preserved for 72 hours via continuous mechanical perfusion with hypertonic citrate purine solution. Upon arrival at the destination, open the cover 14, first release the gas inside the second sealing ring 142, then open the latch or buckle and remove the cover 14.

[0086] Before transplantation, the kidney must be flushed through the renal artery with 1-4°C physiological saline to remove the hypertonic citrate purine solution and prevent the high concentration of potassium ions from this preservation solution from entering the recipient's bloodstream. The specific method is as follows: perfuse the kidney with 4°C physiological saline at a flow rate of approximately 100 mL / min, with a perfusion volume of approximately 500 mL, until the outflow is clear. Then, the transplantation surgery or subsequent experimental procedures can be performed.

[0087] The working principle of this invention is as follows: Passive temperature control principle based on phase change agent The organ preservation tank 2 is a jacketed structure, and the phase change agent 21 filled inside the jacket is the core component for achieving long-distance constant temperature preservation in this invention. The phase change agent 21 utilizes the principle of phase change energy storage, absorbing or releasing a large amount of latent heat when a phase change (solid-liquid phase change or liquid-solid phase change) occurs, thereby maintaining a constant temperature near the phase change temperature point.

[0088] This invention employs a synergistic temperature control strategy of "active cooling + phase change cold storage". When the semiconductor cooler (sidewall cooling mechanism 3) is powered on, according to the Peltier effect, when current flows through a thermocouple composed of two different semiconductor materials, heat is absorbed at the cold end and released at the hot end. The cold end is in close contact with the outer and bottom walls of the organ preservation tank 2, continuously extracting heat from the phase change agent 21 and the interior of the organ preservation tank 2; the hot end dissipates heat to the external environment through the sidewall heat dissipation grooves 13. When the temperature of the phase change agent 21 is higher than the target temperature, the semiconductor cooler activates cooling, lowering the temperature of the phase change agent 21 below the phase change temperature, causing it to undergo a liquid-solid phase change and store a large amount of latent heat. When external heat enters through the wall of the preservation tank 1, the phase change agent 21 absorbs heat through a solid-liquid phase change, maintaining the temperature inside the organ preservation tank 2 near the target temperature. Because the phase change process occurs at a constant temperature, the phase change agent 21 can suppress temperature fluctuations inside the organ preservation tank 2 within a very small range (within ±0.5℃).

[0089] By replacing the phase change agent 21 with one of different phase change temperatures, the same device can be flexibly adapted to the preservation temperature requirements of different tissues and organs. When the target preservation temperature is 35℃ (e.g., isolated pig ovaries), a phase change agent 21 with a phase change temperature of 34–36℃ is selected. During transportation, when the ambient temperature is higher than the phase change temperature, the phase change agent 21 absorbs heat through melting, inhibiting the temperature rise within the organ preservation tank 2; when the ambient temperature is lower than the phase change temperature, the phase change agent 21 releases heat through solidification, inhibiting the temperature drop. The bottom heating mechanism 4 provides compensatory heating when the temperature is too low, and the side-wall cooling mechanism 3 assists in cooling when the temperature is too high.

[0090] When the target preservation temperature is 4℃ (e.g., for eviscerated pig organs), a phase change agent 21 with a phase change temperature of 3-5℃ (e.g., a binary composite solution of n-decyl alcohol / lauric acid) is selected. The side-wall cooling mechanism 3 continuously cools the phase change agent 21 below its phase change temperature, causing it to solidify and store cold. The phase change agent 21 undergoes a phase change at 3-5℃, absorbing heat and stabilizing the temperature inside the organ preservation tank 2 at approximately 4℃. The bottom insulation layer 12 isolates external heat, reducing cold loss.

[0091] Insulating partition 11 isolates the heating device (peristaltic pump 5) from the organ preservation tank 2, preventing the heat generated by the device from affecting the low-temperature environment of the preservation chamber. The insulation material and sealing structure of the preservation box 1 further reduce internal and external heat exchange.

[0092] The infusion circulation principle of this embodiment The peristaltic pump 5 generates negative and positive pressure by periodically squeezing the elastic tube, drawing in and pumping out the hypertonic citrate purine solution from the infusion bag 52. The infusion fluid flows through the supply pipe 51 into the infusion cooling coil 22, which is immersed in the preservation solution 24. After heat exchange with the preservation solution 24 at 4°C, it is cooled to about 4°C and then enters the infusion vessel (such as the renal artery) of the tissue organ 6 through the pipeline.

[0093] After the perfusion fluid enters the organ 6, it flows through the organ's microvascular network, providing the organ's tissue cells with necessary nutrients and oxygen (the perfusion fluid is pre-oxygenated), while simultaneously carrying away waste products generated by cell metabolism. The perfusion fluid flowing out of the organ is led out through the discharge conduit 8 and collected in the waste liquid storage tank 81.

[0094] Continuous cryogenic mechanical perfusion maintains microcirculation in organs in an ex vivo state, avoiding microvascular collapse and endothelial cell damage caused by simple cryopreservation, and effectively prolonging the survival time of organs in vitro.

[0095] This invention achieves real-time monitoring of leakage in the infusion circuit based on the law of conservation of mass. Under normal conditions with no leakage, the inlet flow rate (Q-in) in the supply pipe 51 should be equal to the outlet flow rate (Q-out) in the outlet pipe 8. When a leak occurs at any point in the infusion circuit, some of the infusion agent leaks outside the circuit, causing the outlet flow rate Q-out to be less than the inlet flow rate Q-in.

[0096] The second flow rate sensor 92 on the supply pipe 51 and the first flow rate sensor 91 on the outlet pipe 8 measure Q-in and Q-out in real time, respectively, and transmit the data to the controller. The controller calculates the flow difference ΔQ = Q-in - Q-out. When the absolute value of ΔQ continuously exceeds the preset safety threshold (e.g., 5% of Q-in) and the duration exceeds the set time (e.g., 10 seconds), the controller determines that a leak has occurred in the filling circuit, and then drives the alarm 10 to issue an audible and visual alarm signal, and sends a stop command to the peristaltic pump 5, realizing automatic leak detection, alarm, and protection.

[0097] The multiple physical protection mechanisms of this invention are as follows: Suspension protection: The density of the preservation solution 24 is adjusted to match the density of the tissue / organ 6 (approximately 1.04–1.05 g / cm³). 3 The buoyancy of the organ is similar to that of its weight, which keeps the organ suspended in the preservation solution 24. According to Archimedes' principle, when the buoyancy of the organ is equal to its weight, the organ is in a state of force equilibrium in the liquid, neither floating nor sinking, thus avoiding hard contact between the organ and the bottom or wall of the tank.

[0098] Flexible isolation: A flexible perforated plate 23 is disposed at the bottom of the organ preservation tank 2 to further buffer potential contact between the organ and the bottom of the tank. Through holes 231 ensure free flow of the preservation solution 24 without affecting the contact between the organ's various surfaces and the preservation solution.

[0099] Vibration attenuation: The vibration damping pad 7 is made of high damping material. When vibration energy is transmitted from the storage box 1 to the vibration damping pad 7, the damping material dissipates mechanical energy by converting it into heat energy through internal friction between molecular chains, effectively reducing the vibration amplitude transmitted to the organ storage tank 2.

[0100] Separation and protection: The separator 25 forms a grid-like storage groove to prevent multiple tissues and organs from colliding and squeezing with each other, which is suitable for the classification and preservation of a large number of samples such as pig ovaries.

[0101] The dual sealing principle of this embodiment After the cover 14 is closed, the first sealing ring 141 is compressed and deformed, filling the gap between the cover 14 and the upper surface of the storage box 1, thus achieving an overall seal of the storage box 1. The second sealing ring 142, after being inflated, is embedded in the reserved arc-shaped groove on the upper surface of the organ storage slot 2, independently sealing the upper port of the organ storage slot 2. When it is necessary to open, the gas in the second sealing ring 142 is first released to cause it to contract and disengage from the reserved arc-shaped groove, and then the cover 14 is opened.

[0102] The second sealing ring 142 adopts an inflatable structure, which has the following advantages compared with the traditional fixed sealing ring: the inflation volume is adjustable and the sealing pressure can be flexibly controlled as needed; after expansion, it fits more tightly with the reserved arc groove, resulting in a better sealing effect; after deflation, it shrinks, resulting in low opening resistance and convenient operation.

[0103] The above embodiments are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A device for long-distance transfer and preservation of isolated porcine tissues and organs, characterized in that: Includes a storage box (1), an organ preservation tank (2) disposed in the storage box (1), a cover (14) covering the storage box (1), and a peristaltic pump (5) for perfusing the tissues and organs (6) in the organ preservation tank (2); The storage box (1) is divided into two cavities by an insulating partition (11), which are used to install the organ storage tank (2) and place the peristaltic pump (5), respectively. The organ preservation tank (2) is a jacketed structure, and a phase change agent (21) is filled in the jacket. A side wall cooling mechanism (3) and a bottom heating mechanism (4) are respectively provided on the outer wall and bottom of the organ preservation tank (2). Multiple horizontal and vertical intersecting partitions (25) can also be detachably provided in the organ preservation slot (2) to form a grid-like storage slot for separately preserving animal organs.

2. The device for long-distance transfer and preservation of isolated porcine tissues and organs according to claim 1, characterized in that: A side wall heat dissipation groove (13) is provided on the storage box (1) and on the side wall corresponding to the side wall cooling mechanism (3); a heat insulation layer (12) is provided at the bottom of the storage box (1) and at the bottom corresponding to the bottom heating mechanism (4).

3. The device for long-distance transfer and preservation of isolated porcine tissues and organs according to claim 1, characterized in that: The cover (14) is attached to the storage box (1) by a snap or buckle; A first sealing ring (141) is provided on the cover (14) around the upper end face of the storage box (1); a second sealing ring (142) is provided on the cover (14) around the upper end face of the organ storage slot (2). The second sealing ring (142) is an inflatable and deformable sealing airbag; A reserved arc-shaped groove adapted to the second sealing ring (142) is provided around the upper end face of the organ preservation slot (2); an inflation valve connected to the second sealing ring (142) is provided on the top surface of the cover (14).

4. The device for long-distance transfer and preservation of isolated porcine tissues and organs according to claim 1, characterized in that: A preservation solution (24) is injected into the organ preservation tank (2), the density of which is close to that of the tissue organ (6), so that the tissue organ (6) is suspended in the preservation solution (24).

5. The device for long-distance transfer and preservation of isolated porcine tissues and organs according to claim 4, characterized in that: Inside the storage box (1), there is also a vibration damping pad (7) that supports the bottom and side wall of the organ storage tank (2).

6. The device for long-distance transfer and preservation of isolated porcine tissues and organs according to claim 4, characterized in that: A perforated plate (23) is provided in the organ preservation tank (2), and through holes (231) are evenly spaced on the perforated plate (23). The perforated plate (23) is made of flexible material to prevent the tissue organ (6) from directly contacting the bottom plate of the organ preservation tank (2).

7. The device for long-distance transfer and preservation of isolated porcine tissues and organs according to claim 1, characterized in that: A temperature sensor is provided in the organ preservation tank (2), and the temperature sensor is electrically connected to a controller that controls the side wall cooling mechanism (3) and the bottom heating mechanism (4) respectively.

8. The device for long-distance transfer and preservation of isolated porcine tissues and organs according to claim 1, characterized in that: The organ preservation tank (2) is also provided with an infusion cooling coil (22) and an outlet conduit (8). The infusion cooling coil (22) extends out of the organ preservation tank (2) and is connected to the peristaltic pump (5). Its other end is connected to the inlet blood vessel of the organ (6). The outlet conduit (8) extends out of the organ preservation tank (2) and is connected to a waste liquid storage tank (81). Its other end is connected to the outlet blood vessel of the organ (6). A flow rate sensor is provided on the outlet conduit (8). The flow rate sensor and the peristaltic pump (5) are electrically connected to the controller.

9. The device for long-distance transfer and preservation of isolated porcine tissues and organs according to claim 8, characterized in that: The inlet pipe of the peristaltic pump (5) is connected to the perfusion bag (52), and it is also connected to the perfusion cooling coil (22) through the supply pipe (51) for cooling the perfusion agent in the perfusion bag (52) before it enters the tissue organ (6); the perfusion agent is a hypertonic citrate purine solution.

10. The device for long-distance transfer and preservation of isolated porcine tissues and organs according to claim 9, characterized in that: The flow rate sensor is also provided on the liquid supply pipe (51), and the flow rate sensor is also electrically connected to a controller, which is also connected to an alarm (10).

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