An integrated system for plasma purification and stem cell in vitro secretion regulation

Abstract

This invention provides an integrated system for plasma purification and in vitro stem cell secretion regulation, comprising: a plasma separation and adsorption device for separating plasma from blood and performing plasma replacement removal and / or adsorption treatment before reinfusion; a plasma reverse filtration device for reverse filtration of water-soluble components in plasma; an integrated fiber lumen, wherein the hollow fiber components of the plasma separation and adsorption device and the plasma reverse filtration device are at least partially disposed within the integrated fiber lumen, allowing plasma and filtrate to exchange within the same lumen; and a stem cell secretion regulation device for culturing stem cells in vitro and obtaining their secretions, and introducing the stem cell secretions into the plasma reinfusion pathway for reinfusion into the body; the plasma reverse filtration device simultaneously removes excess water during the reinfusion of stem cell secretions; this invention integrates multiple blood purification modes, effectively avoiding damage to stem cells from plasma toxins while fully utilizing the in vitro secretion regulation function of stem cells.

Description

Technical Field

[0001] This invention relates to the fields of blood purification technology and in vitro stem cell regulation technology, and particularly to an integrated system for plasma purification and in vitro stem cell secretion regulation. Background Technology

[0002] In recent years, blood purification technology has been widely used to remove inflammatory factors and metabolic toxins. Its essence is plasma purification, and common clinical methods include plasma exchange, plasma adsorption, and hemodiafiltration.

[0003] In existing technologies, multiple purification techniques are often used in combination to improve treatment efficacy. However, due to the significant differences in the required blood or plasma flow rates among different blood purification techniques, it is difficult to achieve deep integration of multiple techniques within a single system, increasing the complexity and operational difficulty of combined clinical treatment.

[0004] On the other hand, bioartificial liver technology attempts to combine blood purification with cell therapy (such as stem cells or hepatocytes). Current mainstream methods typically place cells on the outside of a hollow fiber tube, allowing them to exchange substances with the patient's plasma flowing inside. However, in practical applications, because the patient's plasma contains high concentrations of metabolic toxins, these toxins directly toxicize the cells outside the tube, severely inhibiting cell function and shortening their survival time. This makes it difficult for bioartificial livers to achieve the expected therapeutic effects in in vitro secretion regulation and metabolic replacement.

[0005] Therefore, there is an urgent need for an integrated system that combines plasma purification with in vitro secretory regulation of stem cells, integrating multiple blood purification modes, so as to fully utilize the in vitro secretory regulation function of stem cells while effectively avoiding damage to stem cells from plasma toxins. Summary of the Invention

[0006] The purpose of this invention is to provide an integrated system for plasma purification and in vitro stem cell secretion regulation, which aims to solve the technical problems of existing systems being unable to integrate multiple blood purification modes and plasma toxins easily damaging in vitro stem cells, resulting in poor therapeutic effects.

[0007] To achieve the above objectives, the present invention provides an integrated system for plasma purification and in vitro stem cell secretion regulation, comprising:

[0008] A plasma separation and adsorption device is used to separate plasma from blood and to remove and / or adsorb the plasma before returning it to the blood.

[0009] A blood plasma reverse filtration device is used to reverse filter the water-soluble components in the blood plasma;

[0010] An integrated fiber tube cavity is provided in which at least part of the hollow fiber components of the plasma separation and adsorption device and the plasma reverse filtration device are disposed together, so that plasma and filtrate can be exchanged in the same cavity.

[0011] A stem cell secretion regulation device is used to culture stem cells in vitro and obtain their secretions, and to introduce the stem cell secretions into the plasma reinfusion pathway for reinfusion into the body;

[0012] The plasma reverse filtration device is used to simultaneously remove excess water during the reinfusion of stem cell secretions, so as to achieve safe reinfusion of stem cell secretions.

[0013] As a further improvement to the above solution, the plasma separation and adsorption device includes:

[0014] A membrane-type plasma separator is disposed within the integrated fiber lumen.

[0015] A blood collection line connected to the membrane plasma separator is provided with a blood pump, a first blood status monitoring and pressure buffer, and an anticoagulant infusion device.

[0016] A blood return pipeline connected to the membrane plasma separator is provided with a second blood status monitoring and pressure buffer.

[0017] Preferably, the first blood status monitoring and pressure buffer includes a transparent first liquid reservoir and a first pressure measuring tube connected to a pressure sensor, used to observe the blood status in the blood collection tubing, buffer the pulse pressure of blood flow, and monitor the blood pressure in the tubing;

[0018] The second blood status monitoring and pressure buffer includes a transparent second liquid reservoir and a second pressure measuring tube connected to a pressure sensor, used to observe the blood status in the blood return pipeline, buffer the pulse pressure of blood flow, and monitor the blood pressure in the pipeline.

[0019] As a further improvement to the above solution, the integrated fiber tube cavity is equipped with a pressure monitoring interface for real-time monitoring of the pressure inside the cavity.

[0020] As a further improvement to the above solution, the plasma separation and adsorption device further includes:

[0021] The plasma separation pipeline is connected to the membrane plasma separator;

[0022] A plasma pump installed on the plasma separation pipeline;

[0023] The plasma discharge pipeline and the plasma return pipeline, which are connected to the plasma separation pipeline, are connected by a first three-way valve.

[0024] The plasma return pipeline is provided with at least one plasma adsorber and a first heating device in sequence, and is connected to the blood return pipeline; the end of the plasma discharge pipeline is connected to a first waste liquid bag.

[0025] As a further improvement to the above solution, the plasma reverse filtration device includes:

[0026] The reverse plasma filter located within the integrated fiber lumen has one closed end.

[0027] A filtrate inlet pipeline connected to the integrated fiber tube cavity is provided with a filtrate pump, a liquid jug, and a second heating device.

[0028] A switching valve assembly selectively connected to the filtrate inlet line and the blood return line, the switching valve assembly also being connected to the bottom interface of the integrated fiber lumen via a filtrate connection line;

[0029] A filtrate discharge pipeline connected to the reverse plasma filter away from the closed end is provided, and a waste liquid pump is provided on the filtrate discharge pipeline.

[0030] As a further improvement to the above solution, the input end of the filtrate input pipeline can be selectively connected to a plasma bag or a first filtrate bag via a second three-way valve, and the output end of the filtrate discharge pipeline is connected to a second waste liquid bag.

[0031] Both the first filter bag and the second waste liquid bag are equipped with weighing devices to achieve real-time balance control of the inflow and outflow of liquid.

[0032] As a further improvement to the above scheme, the hollow fiber tubes of the membrane plasma separator and the reverse plasma filter are at least partially exposed in the integrated fiber tube lumen.

[0033] As a further improvement to the above scheme, the stem cell secretion regulation device includes:

[0034] A hollow fiber stem cell culture vessel, wherein the outer side of the hollow fiber tube inside the hollow fiber stem cell culture vessel is provided with a stem cell culture area, and the inner side of the hollow fiber tube is a cell-free fluid channel;

[0035] A culture filtrate bag that forms a circulation with the hollow fiber stem cell culture tank and a second filtrate bag connected thereto;

[0036] The filtrate circulation pump, temperature control element and oxygenator are installed on the circulation pipeline;

[0037] A reinfusion pump connected to the bottom of the hollow fiber stem cell culture tank and used to deliver stem cell secretions to the plasma reinfusion pipeline.

[0038] As a further improvement to the above scheme, the hollow fiber tube in the hollow fiber stem cell culture tank is equipped with filter membranes at both ends to prevent stem cells from entering the reinfusion pipeline, while allowing exosomes or microvesicles secreted by stem cells to pass through.

[0039] As a further improvement to the above scheme, the stem cells include mesenchymal stem cells and / or induced pluripotent stem cells, and the stem cells are disposed on the outside of the hollow fiber tube in the form of three-dimensional culture microspheres or adherent cells.

[0040] As a further improvement to the above scheme, the stem cell secretion regulation device can operate as an independent module or in conjunction with the plasma separation and adsorption device and the plasma reverse filtration device.

[0041] As a further improvement to the above scheme, the pumping speed of the blood pump is 100-150 mL / min;

[0042] The pumping speed of the plasma pump is 20–30 mL / min;

[0043] The pumping speed of both the filtrate pump and the waste liquid pump is 20–100 mL / min;

[0044] The pumping speed of the return pump is 2 to 5 mL / min.

[0045] Because the present invention adopts the above technical solutions, the beneficial effects of this application are as follows:

[0046] This invention discloses an integrated system for plasma purification and in vitro stem cell secretion regulation. Through an integrated fiber tubing cavity, it achieves a compact integration of multiple purification modes, improving operational convenience and space utilization. Specifically, this invention firstly, at least partially integrates the hollow fiber components of the plasma separation and adsorption device and the plasma reverse filtration device within the integrated fiber tubing cavity, allowing the core components of the two purification modes to share the same cavity space. On the one hand, this avoids the dispersed arrangement of two independent devices, significantly reducing the system volume and facilitating space utilization in clinical settings. On the other hand, the exchange of plasma and filtrate is completed within the same cavity, reducing pipeline connection points, lowering the risk of leakage and pipeline blockage, and simplifying the operation process, thus improving the convenience of clinical use.

[0047] Secondly, the integrated system provided by this invention allows for the initial separation and replacement of plasma, followed by a combined process of plasma adsorption and reverse filtration. During plasma exchange, the return pump at the bottom of the hollow fiber stem cell culture tank, used for the reinfusion of stem cell secretions, is not operational, thus creating a closed loop of the culture medium inside and outside the hollow fiber tubes of the stem cell culture tank, increasing the concentration of stem cell secretions in the culture medium. During plasma adsorption and reverse filtration, the culture medium in the hollow fiber stem cell culture tank is unidirectionally reinfused into the plasma through the return pump at its lower end, thereby preventing plasma from entering the hollow fiber stem cell culture tank. This avoids damage to stem cells from plasma toxins at the source and ensures the activity and secretory function of stem cells, laying the foundation for the subsequent therapeutic effects of stem cell secretions.

[0048] Furthermore, stem cell secretions typically contain excess water generated during the culture process, such as residual culture medium and metabolic water. Direct reinfusion can increase the patient's blood volume overload, potentially inducing adverse reactions such as heart failure and pulmonary edema. The plasma reverse filtration device of this invention activates simultaneously during stem cell secretion reinfusion, precisely removing excess water from the entering secretions without requiring additional dehydration equipment. This design simplifies the reinfusion process, avoiding the secondary processing of "culture first, then separate dehydration," while ensuring an appropriate volume of reinfused fluid, effectively mitigating the clinical risk of volume overload and improving the safety of stem cell secretion reinfusion.

[0049] This invention integrates plasma purification (plasma replacement, adsorption, and reverse filtration) with stem cell secretion regulation (culture and secretion fluid reinfusion) into a single system, achieving a closed-loop process of "plasma toxin removal → stem cell protection → secretion fluid generation → safe reinfusion." After separation, replacement, and adsorption to remove toxins, the plasma is directly reinfused to maintain internal environmental stability. The stem cell secretion fluid, after reverse filtration and dehydration, is safely reinfused, and its effective components synergistically enhance the therapeutic effect of the purified plasma. This integrated design avoids the errors associated with switching between multiple devices, optimizes the continuity of the treatment process, and ultimately improves the overall precision and effectiveness of the treatment. Attached Figure Description

[0050] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0051] Figure 1 This is a schematic diagram of the structural principle of an integrated system for plasma purification and in vitro secretion regulation of stem cells disclosed in this invention;

[0052] Figure 2 This is a schematic diagram of the structural principle of the stem cell secretion regulation device disclosed in this invention.

[0053] Figure label:

[0054] 1. Plasma separation and adsorption device; 11. Membrane plasma separator; 12. Blood collection tubing; 121. Blood pump; 122. First blood status monitoring and pressure buffer; 123. Anticoagulant infusion device; 13. Plasma separation tubing; 131. Plasma pump; 14. First three-way valve; 15. Plasma discharge tubing; 151. First waste bag; 16. Plasma return tubing; 17. Plasma adsorber; 18. First heating device;

[0055] 2. Reverse plasma filtration device; 20. Reverse plasma filter; 21. Filtrate inlet line; 210. Filtrate pump; 211. Liquid reservoir; 212. Second heating device; 213. Plasma bag; 214. First filtrate bag; 215. Second three-way valve; 22. Switching valve assembly; 23. Filtrate outlet line; 231. Waste liquid pump; 232. Second waste liquid bag; 24. Weighing device; 25. Blood return line; 251. Second blood status monitoring and pressure buffer; 26. Filtrate connection line;

[0056] 3. Integrated fiber optic tubing; 31. Pressure monitoring interface;

[0057] 4. Stem cell secretion regulation device; 40. Hollow fiber stem cell culture tank; 41. Culture filtrate bag; 42. Filtrate circulation pump; 43. Return pump; 44. Temperature control element; 45. Oxygenator; 46. Second filtrate bag; 47. Clamps. Detailed Implementation

[0058] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0059] It should be noted that all directional indicators (such as up, down, etc.) in the embodiments of the present invention are only used to explain the relative positional relationship and movement of each component in a certain specific posture (as shown in the figure). If the specific posture changes, the directional indicator will also change accordingly.

[0060] Furthermore, in this invention, descriptions involving "first," "second," etc., are for descriptive purposes only and should not be construed as indicating or implying their relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined with "first" or "second" may explicitly or implicitly include at least one of that feature.

[0061] Furthermore, the technical solutions of the various embodiments of the present invention can be combined with each other, but only if they are based on the ability of those skilled in the art to implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such combination of technical solutions does not exist and is not within the scope of protection claimed by the present invention.

[0062] See Figure 1 and Figure 2 This invention provides an integrated system for plasma purification and in vitro stem cell secretion regulation, comprising:

[0063] The plasma separation and adsorption device 1 is used to separate plasma from blood and perform displacement removal and adsorption treatment on the plasma before reintroduction. Its core component is a hollow fiber membrane module, which can accurately separate plasma from blood while retaining blood cell components to prevent loss. After plasma separation, plasma displacement is performed first, followed by selective adsorption treatment of non-water-soluble toxins in the plasma. After adsorption, the plasma is reintroduced into subsequent pathways.

[0064] The plasma reverse filtration device 2 is used to reverse filter the water-soluble components in the plasma; it is also equipped with hollow fiber components, which can reverse filter the water-soluble small molecule toxins and electrolytes in the plasma after separation by the membrane plasma separator, thereby achieving deep purification of the plasma. Compared with traditional standalone filtration devices, this device, through its integrated layout with the plasma separation adsorption device 1, can form a synergistic "adsorption-filtration" multi-mode blood purification system.

[0065] The integrated fiber optic lumen 3 is a cylindrical or other shaped (e.g., figure-eight shaped cross-section) sealed cavity. The hollow fiber assembly of the plasma separation and adsorption device 1 and the hollow fiber assembly of the plasma reverse filtration device 2 are partially overlapped along the axial direction within the integrated fiber optic lumen 3. This allows the effluent after plasma separation and the filtrate from the reverse filtration to exchange substances within the same cavity, eliminating the need for additional connecting pipes to achieve fluid communication between the two devices. This integrated design significantly shortens the plasma transport path, reduces the plasma's residence time outside the body, and simultaneously completes plasma adsorption and filtration to remove water-soluble toxins. This reduces the cumulative effect of residual toxins in the plasma and minimizes the risk of contamination from pipe connections. Furthermore, the compact structure enhances the system's portability, making it suitable for bedside clinical applications.

[0066] The stem cell secretion regulation device 4 is used to culture stem cells in vitro and obtain their secretions, and to introduce the stem cell secretions into the plasma reinfusion pathway for reinfusion into the body.

[0067] The plasma reverse filtration device 2 plays a role in the reinfusion of stem cell secretions. After the stem cell secretions are mixed with the purified plasma, they enter the human body. The plasma is separated and passes through the filter membrane of the plasma reverse filtration device 2 to remove excess water entering the human body, thus avoiding adverse reactions caused by excessive reinfusion of secretions or abnormal osmotic pressure, and achieving safe reinfusion of stem cell secretions.

[0068] Each device is structurally integrated through an integrated fiber lumen 3, forming an integrated operation process of "plasma separation and adsorption - reverse filtration and purification - stem cell secretion fluid reinfusion and water regulation". It integrates multiple blood purification modes, improves plasma purification efficiency and comprehensiveness, and at the same time, with the synchronous water regulation function of the plasma reverse filtration device 2, ensures the safe reinfusion of stem cell secretion fluid, effectively solving the core problems existing in the current technology.

[0069] In a preferred embodiment, the plasma separation and adsorption device 1 includes:

[0070] The membrane plasma separator 11 uses a hollow fiber membrane module, such as a membrane pore size of 0.2-0.6μm and a separation efficiency of ≥95%. It is fixed on a support inside the integrated fiber lumen 3. Its blood inlet is connected to the patient's vascular access through the blood collection line 12, and the plasma outlet is connected to the adsorption column (with built-in activated carbon / resin). The blood outlet is returned to the patient's body through the blood reinfusion line 25.

[0071] The blood collection tubing 12, from the patient end to the inlet end of the membrane plasma separator 11, is equipped with a blood pump 121, a first blood status monitoring and pressure buffer 122, and an anticoagulant infusion device 123. The anticoagulant infusion device 123 is preferably a heparin pump with an infusion rate of 1-5 U / kg·h.

[0072] The blood reinfusion line 25 is equipped with a second blood status monitoring and pressure buffer 251 from the outlet end of the membrane plasma separator 11 to the patient end.

[0073] Preferably, the first blood status monitoring and pressure buffer 122 includes a transparent first liquid reservoir and a first pressure measuring tube connected to a pressure sensor. The transparent first liquid reservoir is made of medical-grade polycarbonate, with the first pressure measuring tube vertically connected to its side wall, and the pressure sensor connected to the end of the pressure measuring tube. When blood is delivered to the first liquid reservoir by the blood pump 121, the transparent reservoir wall allows for direct observation of whether there are air bubbles, thrombi, or hemolysis in the blood. The operator can promptly stop the blood pump 121 and handle any abnormalities. The buffer chamber structure of the liquid reservoir 211 can alleviate the pulse pressure of the blood pump 121 and prevent the pipeline from being damaged due to sudden pressure changes. The first pressure measuring tube transmits pressure signals to the control system in real time, maintaining the arterial pressure within the normal range. If the pressure is >180 mmHg, an alarm is triggered to prevent blood reflux or membrane contamination.

[0074] The second blood status monitoring and pressure buffer 251 includes a transparent second reservoir and a second pressure measuring tube connected to a pressure sensor. The transparent second reservoir is made of medical-grade polycarbonate, with the second pressure measuring tube vertically connected to its side wall. The pressure sensor is connected to the end of the pressure measuring tube. Blood cells (red blood cells, white blood cells, and platelets) separated by the membrane separator enter the second reservoir, allowing observation of blood cell aggregation, such as rouleaux formation of red blood cells due to hypercoagulability, thus avoiding the reinfusion of blood containing a large number of microthrombi. The buffering function reduces the impact pressure of blood cells on the reinfusion tubing, minimizing damage to the blood vessel wall. The second pressure measuring tube monitors venous pressure; if the pressure is <10 mmHg, the reinfusion is immediately stopped to ensure patient safety.

[0075] The anticoagulant infusion device 123 is located upstream of the first infusion vessel. Heparin solution (or other anticoagulant) enters the first infusion vessel simultaneously with the blood, mixes thoroughly with the blood, and then flows to the membrane separator. The buffering effect of the first infusion vessel ensures that the anticoagulant and blood are mixed evenly, avoiding the risk of bleeding due to excessively high local anticoagulant concentration or the risk of clotting in the tubing due to excessively low concentration. At the same time, by observing the state of the blood in the first infusion vessel, if no blood clots adhere to the vessel wall, the anticoagulant effect can be indirectly verified without the need for additional sampling and testing.

[0076] In a preferred embodiment, the integrated fiber optic lumen 3 has a pressure monitoring interface 31 on its side wall. The interface is fitted with a medical-grade silicone sealing plug and connected to an external pressure sensor. The pressure sensor signal line is connected to the system's main control unit, which can display the pressure value within the lumen in real time and plot a pressure curve. The main control unit presets a pressure safety threshold, such as a normal operating range of 50-180 mmHg. When the lumen pressure approaches the upper limit of the threshold, it automatically reduces the speed of the blood pump 121 to reduce transmembrane pressure. If the pressure remains abnormal, it prompts for replacement of the membrane assembly or troubleshooting of the tubing to prevent irreversible damage to the membrane assembly or plasma leakage due to excessive pressure. By setting a pressure monitoring interface 31 in the integrated fiber optic lumen 3 and linking it with the pressure sensor and main control unit, real-time visualization and dynamic control of the pressure within the lumen are achieved, reducing the risk of membrane assembly blockage or rupture. The automatic alarm and speed reduction function in case of abnormal pressure prevents plasma leakage or tubing detachment caused by sudden pressure changes, improving the safety and stability of equipment operation. Continuous recording of pressure data can also be used for postoperative follow-up, providing a basis for optimizing treatment plans.

[0077] In a preferred embodiment, based on the original plasma separation and adsorption device 1, a plasma separation pipeline 13, a plasma pump 131, a plasma discharge pipeline 15, a plasma return pipeline 16, and a first three-way valve 14 are further added to form a more flexible dual-path plasma processing system. The specific connection relationships are shown below:

[0078] The plasma separation pipeline 13 is connected at one end to the plasma outlet of the membrane plasma separator 11, and at the other end it is divided into two branches: one branch is connected to the plasma return pipeline 16 via the first three-way valve 14, and the other branch is connected to the plasma discharge pipeline 15; a plasma pump 131, such as a peristaltic pump, is installed on the pipeline to control the rate at which plasma is output from the separator.

[0079] The plasma reinfusion pipeline 16, starting from the first three-way valve 14, connects at least one plasma adsorber 17 and a first heating device 18 in series, and finally connects to the blood reinfusion pipeline 25, that is, the purified plasma is mixed with blood cells and reinfused into the patient; in this embodiment, two plasma adsorbers 17 are connected in series, and the first heating device 18 adopts a constant temperature water tank or electric heating wire, and the temperature is controlled within the range of 35-38℃.

[0080] The plasma discharge line 15 extends from the other side of the first three-way valve 14 and is connected to the first waste liquid bag 151 at the end. It is used to collect plasma components that need to be discarded, such as plasma with high toxins and high inflammatory factors.

[0081] In clinical practice, the workflow of the plasma separation and adsorption device 1 can be divided into three stages: plasma diversion, differential treatment, and safe reinfusion.

[0082] Plasma diversion: After the membrane plasma separator 11 separates the plasma from the blood, the plasma pump 131 starts and pumps the plasma into the plasma separation pipeline 13; at this time, the first three-way valve 14 is in the "dual-path selection" state, and the plasma is divided into two streams:

[0083] The first stream (waste path): The plasma flows into the first waste bag 151 through the plasma discharge pipeline 15. It mainly consists of high-concentration toxic plasma, such as bilirubin >200μmol / L in the plasma of patients with liver failure, or endotoxin >0.5EU / mL in the plasma of patients with sepsis. This type of high-concentration toxic plasma is directly discarded. At the same time, the input end of the filtrate input pipeline 21 of the plasma reverse filtration device 2 is connected to the plasma bag 213 through the second three-way valve 215. The switching valve assembly 22 connects the end of the filtrate input pipeline 21 with the blood return pipeline 25. The filtrate pump 210 is started to return the plasma from the plasma bag 213 to complete the plasma replacement.

[0084] Second path (adsorption path): After plasma exchange is completed, adjust the first three-way valve 14 so that the plasma separation pipeline 13 no longer passes through the plasma discharge pipeline 15, but enters the plasma adsorber 17 through the plasma return pipeline 16, and is returned after adsorption.

[0085] The plasma entering the adsorption pathway passes through the following sequentially:

[0086] The plasma adsorber 17 uses activated carbon to adsorb liver failure toxins, such as bilirubin and bile acids, and resin to adsorb macromolecular toxins and immune complexes, such as endotoxins, inflammatory factors, rheumatoid factors, and antinuclear antibodies.

[0087] The first heating device 18 raises the plasma temperature from room temperature to 35-38°C, making the plasma temperature close to the human body temperature, thereby avoiding adverse reactions such as vasoconstriction and chills caused by direct reinfusion of low-temperature plasma, while maintaining the activity of coagulation factors in the plasma.

[0088] The heated purified plasma mixes with blood cells (from the blood outlet of the membrane plasma separator 11) in the blood reinfusion line 25 at the confluence node, forming a complete blood component of "blood cells + purified plasma", which is then reinfused into the patient. During this process, the first blood status monitor and pressure buffer 122 can observe the homogeneity of the mixed blood, such as the absence of plasma and blood cell stratification; the second blood status monitor and pressure buffer 251 monitors the reinfusion pressure.

[0089] Through the improvements to the pipelines and valves mentioned above, the plasma separation and adsorption device 1 achieves refined purification through "on-demand diversion and differentiated treatment," sequentially completing plasma exchange and plasma adsorption, while reducing the risk of adverse reactions during patient reinfusion.

[0090] In a preferred embodiment, the plasma reverse filtration device 2 includes:

[0091] The reverse plasma filter 20 is installed inside the integrated fiber lumen 3 and includes a hollow fiber membrane assembly, one end of which is a closed end and the other end is an open end.

[0092] The filtrate inlet pipeline 21 is connected to either a plasma bag 213 or a first filtrate bag 214 via a second three-way valve 215. A filtrate pump 210, a transparent liquid reservoir 211, and a second heating device 212 are sequentially arranged on the pipeline. The plasma bag 213 stores fresh plasma to be reinfused, and the first filtrate bag 214 stores blood filtrate. Preferably, in this embodiment, the filtrate pump 210 is a peristaltic pump. The second heating device 212 uses a constant temperature water tank or an electric heating wire, with the temperature controlled within the range of 36-37°C to match human body temperature.

[0093] The switching valve assembly 22 is located between the end of the filtrate inlet line 21 and the blood return line 25. The switching valve assembly 22 is also connected to the bottom interface of the integrated fiber lumen 3 through the filtrate connection line 26. Preferably, it is a three-way electromagnetic reversing valve, which can realize the path switching of "fresh plasma inlet → blood return" or "blood filtrate inlet → integrated fiber lumen 3 → filtrate outlet line 23".

[0094] The filtrate discharge pipeline 23 has its input end connected to the open end of the reverse plasma filter 20, and its output end connected to the second waste bag 232 via the waste pump 231.

[0095] The metering device includes an electronic weighing device 24 at the bottom of both the first filter bag 214 and the second waste liquid bag 232. The signal is connected to the main control unit of the system for real-time calculation of the difference between the inflow and outflow of liquid.

[0096] In clinical applications, the plasma reverse filtration device 2 is mainly used to simultaneously and reversely remove water-soluble toxins from the plasma and excess water from the stem cell secretion fluid during plasma separation and adsorption. The specific process of the plasma reverse filtration mode is as follows:

[0097] Path switching: The second three-way valve 215 connects the input end of the filtrate input pipeline 21 to the first filtrate bag 214, which is pre-filled with blood filtrate. The switching valve assembly 22 switches to the "filtrate input → filtrate connection pipeline 26" direction.

[0098] Reverse filtration of filtrate: When filtrate pump 210 is activated, the blood filtrate is pumped from the first filtrate bag 214 through the filtrate connecting pipe 26 into the integrated fiber lumen 3, where it mixes with the plasma separated by the membrane plasma separator 11. Part of the mixture enters the adsorption path and is reinfused into the body, while the other part mixes with the high concentration of water-soluble toxins diffused and filtered out by the membrane plasma separator 11 and enters the reverse plasma filter 20 in reverse. From the output end, it is pumped into the second waste bag 232 via waste pump 231. By adjusting the speeds of filtrate pump 210 and waste pump 231, excess water in the reinfused fluid of stem cell secretion can be removed. For example, when the reinfusion pump 43 of the stem cell secretion regulation device 4 has a pumping speed of 5 ml / min, setting the pumping speed of filtrate pump 210 to 90 ml / min and the pumping speed of waste pump 231 to 95 ml / min can achieve a balance between the amount of water entering and leaving the body.

[0099] In a preferred embodiment, the stem cell secretion regulation device 4 is based on the hollow fiber stem cell culture tank 40, and forms a closed-loop system by combining the filtrate circulation and secretion fluid reinfusion pipeline, as shown in the following specific structure:

[0100] Hollow fiber stem cell culture tank 40 is a medical-grade sealed container with multiple sets of hollow fiber tubes inside. The outer side of the hollow fiber tube is the stem cell culture area, and the inner side is a cell-free fluid channel. Both ends of the fiber tube are equipped with filter membranes, which only allow liquids, exosomes or microvesicles to pass through, and prevent stem cells and debris from entering the channel.

[0101] The filtrate circulation system connects the cell-free fluid channel of the culture tank to a culture filtrate bag 41. The inlet of the culture filtrate bag 41 is also connected to a second filtrate bag 46 via a pipe and clamp 47. The second filtrate bag 46 stores blood filtrate or other stem cell culture medium that can enter the human body. The outlet of the culture filtrate bag 41 is connected to a filtrate circulation pump 42. The outlet of the filtrate circulation pump 42 flows back to the hollow fiber stem cell culture tank 40 via a temperature control element 44 and an oxygenator 45, forming a closed loop. The filtrate circulation pump 42 drives the filtrate to circulate inside and outside the hollow fiber channel to increase the concentration of stem cell secretions in the liquid inside the hollow fiber channel, preparing for the reinfusion of high-concentration stem cell secretions.

[0102] The secretion fluid return pipeline has a drainage port at the bottom of the culture tank, which is connected to the plasma return pipeline 16 via the return pump 43. This is used to transport the active components secreted by stem cells back into the body. Driven by the return pump 43, the stem cell filtrate in the hollow fiber stem cell culture tank 40 flows through the plasma return pipeline 16 into the blood return pipeline 25, returning to the patient's body. Preferably, the speed of the filtrate circulation pump 42 is greater than that of the return pump 43, typically set to five times the latter.

[0103] In clinical applications, the workflow of the stem cell secretion regulation device 4 revolves around "dynamic culture - secretion fluid collection - safe reinfusion," as detailed below:

[0104] 1. Dynamic culture of stem cells and circulation of filtrate:

[0105] Inoculation and initialization: The expanded mesenchymal stem cell suspension is injected into the outer culture area of ​​the culture tank. The blood filtrate / culture medium from the second filtrate bag 46 is added to the culture filtrate bag 41. The circulation is started by the filtrate circulation pump 42, so that the filtrate slowly circulates from the outside to the inside of the hollow fiber tube.

[0106] Dynamic Culture and Nutrient Exchange: During culture, the filtrate circulation pump 42 maintains a constant circulation rate. Stem cell secretions from the outer culture zone flow into the inner cell-free channel with the culture filtrate and then flow back to the culture filtrate bag 41. Simultaneously, fresh nutrients from the culture filtrate bag 41 are continuously pumped into the culture zone by the filtrate circulation pump 42, achieving a microcirculation simulation of "dynamic nutrient supply - secretion accumulation." Additionally, blood filtrate / culture medium from the second filtrate bag 46 can be added to this circulation to provide fresh nutrients for the stem cells. This process avoids the "nutrient depletion - metabolite accumulation" problem of traditional static culture, effectively improving stem cell survival rate and increasing secretion concentration.

[0107] 2. Collection of secretions and interception of stem cells:

[0108] Secretion drainage: After culturing for the preset time, start the reinfusion pump 43 to pump the secretion from the bottom of the culture tank into the plasma reinfusion line 16; the filter membranes at both ends of the hollow fiber tube (different membrane pore sizes can be selected according to needs, usually 0.45μm) can effectively intercept stem cells and cell debris >0.45μm, allowing only secretion products <0.45μm to pass through, thus avoiding the risk of tumorigenesis or immune rejection of stem cell reinfusion into the body.

[0109] Filate circulation maintenance: During the drainage of secretions, the filtrate circulation pump 42 runs continuously to keep the culture area moist and nutrient-supplied, preventing the remaining stem cells from dying due to dryness or lack of nutrition.

[0110] 3. Synergy with plasma reinfusion pathways:

[0111] After the reinfusion pump 43 injects the secretion into the plasma reinfusion line 16, the second blood status monitoring and pressure buffer 251 can observe the uniformity of the mixing of the secretion and plasma. If further dehydration is required, the reinfusion line can be connected to the reverse filter through the switching valve assembly 22 of the plasma reverse filtration device 2 to remove excess water simultaneously.

[0112] As a preferred embodiment, based on the hollow fiber stem cell culture vessel 40, the type of stem cells and the specific culture method in the culture area outside the hollow fiber tube are further clarified to adapt to different treatment needs and optimize the secretion effect:

[0113] Stem cell type: Mesenchymal stem cells (MSCs) and / or induced pluripotent stem cells (iPSCs) are selected. MSCs are derived from bone marrow, adipose tissue, or umbilical cord and have low immunogenicity and directed differentiation potential; iPSCs are obtained by reprogramming somatic cells and can differentiate into various functional cells, making them suitable for the treatment of complex diseases.

[0114] Culture format: Stem cells are distributed in the culture zone outside the hollow fiber tube as three-dimensional culture microspheres or adherent monolayers.

[0115] Three-dimensional cultured microspheres: Using microfluidic 3D printing technology, stem cell microspheres with a diameter of 200-500μm can be prepared using FDA-approved excipients (such as matrix materials such as collagen / alginate), and then transferred to the outside of hollow fiber tubes;

[0116] Adherent monolayer: Stem cells are directly seeded onto the surface of a hollow fiber tube coated with matrix gel, allowing them to adhere and grow into a monolayer of cells.

[0117] In a preferred embodiment, the stem cell secretion regulation device 4 can operate as an independent module or in conjunction with the plasma separation and adsorption device 1 and the plasma reverse filtration device 2.

[0118] The stem cell secretion regulation device 4 constitutes a closed-loop system, including a hollow fiber stem cell culture tank 40, a filtrate circulation pump 42, a culture filtrate bag 41, a reinfusion pump 43, and an independent power supply / control unit. It is directly connected to the patient's intravenous access through a dedicated infusion line, without relying on the plasma separation and adsorption device 1 or the reverse filtration device.

[0119] Collaborative operation mode: The filtrate circulation pipeline of the stem cell secretion regulation device 4 is connected to the plasma return pipeline 16 of the plasma separation and adsorption device 1 and the filtrate input pipeline 21 of the plasma reverse filtration device 2 through a quick-connect connector. Its control unit can communicate with the system main control unit to achieve data sharing and synchronous regulation.

[0120] In a preferred embodiment, the blood pump 121 is installed in the blood collection tubing 12 to drive blood from the patient's vascular access into the membrane plasma separator 11, with the pump speed limited to 100-150 mL / min.

[0121] The plasma pump 131 is installed in the plasma separation pipeline 13 and drives the separated plasma from the membrane plasma separator 11 into the plasma reinfusion / discharge pipeline. The pump speed is limited to 20-30 mL / min.

[0122] The filtrate pump 210 and the waste liquid pump 231 are respectively installed in the filtrate inlet pipe 21 and the filtrate outlet pipe 23, driving the filtrate to circulate or be discharged in the reverse plasma filter 20, with a pump speed of 20-100 mL / min.

[0123] The reinfusion pump 43 is located between the bottom of the hollow fiber stem cell culture tank 40 and the plasma reinfusion pipeline 16, driving the secretion fluid to be reinfused into the body, with the pump speed limited to 2-5 mL / min.

[0124] To further illustrate the inventive concept of the present invention, the operation of the plasma separation and adsorption device 1 will be described in detail. Specifically, the plasma separation and adsorption device 1 operates in two stages: the first stage is the plasma replacement stage, and the second stage is the plasma adsorption stage.

[0125] During plasma exchange therapy using the plasma separation and adsorption device 1, the patient's blood is introduced into the membrane plasma separator 11 at a rate of 100-150 ml / min via the blood collection line 12 and driven by the blood pump 121. Driven by the plasma pump 131, the plasma is separated from the membrane plasma separator 11 and enters the integrated fiber lumen 3. It then flows from the plasma separation line 13, through the first three-way valve 14, into the waste plasma bag via the plasma discharge line 15. During this stage, the plasma return line 16 is kept closed by adjusting the direction of the first three-way valve 14. Simultaneously, the filtrate pump 210 drives the fresh plasma in the plasma bag 213 at a constant rate of 20-30 ml / min from the filtrate inlet line 21 into the blood return line 25, returning it to the patient's body, thus completing the plasma exchange therapy. During this stage, the second three-way valve 215 is adjusted to connect the filtrate inlet line 21 to the plasma bag 213, but not to the first filtrate bag 214; the direction of the switching valve assembly 22 is adjusted to connect the filtrate inlet line 21 to the blood return line 25, but not to the filtrate connection line 26. During plasma exchange, the waste fluid pump 231 remains stationary and off.

[0126] When the plasma separation and adsorption device 1 performs plasma adsorption therapy, the first three-way valve 14 is adjusted to connect the plasma separation pipeline 13 with the plasma return pipeline 16, and the plasma discharge pipeline 15 is cut off to ensure that the plasma separated by the plasma separation pipeline 13 passes through the two plasma adsorbers 17 in sequence and then flows from the plasma return pipeline 16 into the blood return pipeline 25, returning to the patient's body to complete the plasma adsorption.

[0127] The plasma reverse filtration device 2 simultaneously performs plasma reverse filtration treatment while the plasma separation and adsorption device 1 is performing plasma adsorption treatment. The second three-way valve 215 is adjusted so that the filtrate input line 21 is not connected to the plasma bag 213, but is connected to the first filtrate bag 214; the direction of the switching valve assembly 22 is adjusted so that the filtrate input line 21 is not connected to the blood return line 25, but is connected to the filtrate connection line 26. Driven by the filtrate pump 210, the filtrate in the first filtrate bag 214 enters from the bottom of the integrated fiber tube cavity 3 through the filtrate input pipe 21 and fills the space of the integrated fiber tube cavity 3. The plasma and water-soluble toxins in the membrane plasma separator 11 enter the integrated fiber tube cavity 3 through filtration, diffusion and other means and mix with the filtrate input through the filtrate input pipe 21. A portion of the water-soluble toxins in the mixture enters the reverse plasma filter 20 under the drive of the waste liquid pump 231. Preferably, the reverse plasma filter 20 is composed of blood filtration hollow fiber tubes with a membrane pore size of 3 nm. This membrane only allows small and medium-sized water-soluble toxins to pass through. Albumin cannot pass through, and exosomes and vesicles secreted by stem cells cannot pass through (usually with a diameter greater than 30 nm). The bottom of the reverse plasma filter 20 is closed, and the top is connected to the filtrate discharge line 23. The waste liquid pump 231 drives the filtrate in the filtrate discharge line 23 into the waste liquid bag, and the pump speed is 50-100 ml / min. The plasma in the mixture and another part of the filtrate are driven by the plasma pump through the plasma separation line 13, the plasma adsorber 17 and the plasma return line 16 into the blood return line 25 and returned to the patient's body.

[0128] The above are merely preferred embodiments of the present invention and do not limit the patent scope of the present invention. Any equivalent structural transformations made using the contents of the present invention's specification and drawings under the inventive concept of the present invention, or direct or indirect applications in other related technical fields, are within the patent protection scope of the present invention.

Claims

1. An integrated system for plasma purification and in vitro stem cell secretion regulation, characterized in that, include: A plasma separation and adsorption device is used to separate plasma from blood and to remove and / or adsorb the plasma before reintroducing it. A blood plasma reverse filtration device is used to reverse filter the water-soluble components in the blood plasma; An integrated fiber tube cavity is provided in which at least part of the hollow fiber components of the plasma separation and adsorption device and the plasma reverse filtration device are disposed together, so that plasma and filtrate can be exchanged in the same cavity. A stem cell secretion regulation device is used to culture stem cells in vitro and obtain their secretions, and then introduce the stem cell secretions into the plasma reinfusion pathway for reinfusion into the body. The plasma separation and adsorption device includes: a membrane plasma separator disposed within the integrated fiber lumen; a blood collection pipeline connected to the membrane plasma separator; a blood return pipeline connected to the membrane plasma separator; and a plasma separation pipeline connected to the integrated fiber lumen; as well as a plasma discharge pipeline and a plasma return pipeline connected to the plasma separation pipeline, which are connected by a first three-way valve; the plasma return pipeline is connected to the blood return pipeline, and the membrane plasma separator includes a hollow fiber membrane module; The plasma reverse filtration device includes: A reverse plasma filter is disposed within the integrated fiber lumen, the reverse plasma filter comprising a hollow fiber membrane assembly, one end of which is a closed end; A filtrate inlet line connected to the integrated fiber tubing; A switching valve assembly is selectively connected to the filtrate inlet line and the blood return line. The switching valve assembly is also connected to the bottom interface of the integrated fiber tube cavity via a filtrate connection line. The hollow fiber tubes of the membrane plasma separator and the reverse plasma filter are at least partially exposed in the integrated fiber tube cavity, so that the effluent after plasma separation and the filtrate of the reverse plasma filter complete the exchange of substances in the same cavity. The stem cell secretion regulation device includes a hollow fiber stem cell culture tank and a reinfusion pump connected to the bottom of the hollow fiber stem cell culture tank and used to deliver stem cell secretions to the plasma reinfusion pipeline; the plasma reverse filtration device is used to simultaneously remove excess water during the reinfusion of stem cell secretions in order to achieve safe reinfusion of stem cell secretions.

2. The integrated system for plasma purification and in vitro stem cell secretion regulation according to claim 1, characterized in that, The blood collection line is equipped with a blood pump, a first blood status monitoring and pressure buffer, and an anticoagulant infusion device; the blood return line is equipped with a second blood status monitoring and pressure buffer.

3. The integrated system for plasma purification and in vitro secretion regulation of stem cells according to claim 1, characterized in that, The integrated fiber tube cavity is equipped with a pressure monitoring interface for real-time monitoring of the pressure inside the cavity.

4. The integrated system for plasma purification and in vitro stem cell secretion regulation according to claim 2, characterized in that, The plasma separation and adsorption device further includes: A plasma pump is installed on the plasma separation pipeline; wherein, at least one plasma adsorber and a first heating device are sequentially provided on the plasma return pipeline; and a first waste liquid bag is connected to the end of the plasma discharge pipeline.

5. The integrated system for plasma purification and in vitro stem cell secretion regulation according to claim 2, characterized in that, The plasma reverse filtration device also includes: A filtrate discharge pipeline connected to the reverse plasma filter away from the closed end is provided. A waste liquid pump is provided on the filtrate discharge pipeline. A filtrate pump, a liquid jug, and a second heating device are provided on the filtrate input pipeline.

6. The integrated system for plasma purification and in vitro stem cell secretion regulation according to claim 5, characterized in that, The inlet of the filtrate inlet pipeline is connected to a plasma bag or a first filtrate bag via a second three-way valve, and the outlet of the filtrate outlet pipeline is connected to a second waste bag. Both the first filter bag and the second waste liquid bag are equipped with weighing devices to achieve real-time balance control of the inflow and outflow of liquid.

7. The integrated system for plasma purification and in vitro stem cell secretion regulation according to claim 1, characterized in that, The stem cell secretion regulation device also includes: A culture filtrate bag that forms a circulation with the hollow fiber stem cell culture tank and a second filtrate bag connected thereto; The filtrate circulation pump, temperature control element and oxygenator are installed on the circulation pipeline; The hollow fiber stem cell culture tank has a stem cell culture area on the outside of the hollow fiber tube and a cell-free fluid channel on the inside of the hollow fiber tube.

8. The integrated system for plasma purification and in vitro stem cell secretion regulation according to claim 7, characterized in that, The hollow fiber tube inside the hollow fiber stem cell culture tank is equipped with filter membranes at both ends to prevent stem cells from entering the reinfusion line, while allowing exosomes or microvesicles secreted by stem cells to pass through.

9. The integrated system for plasma purification and in vitro stem cell secretion regulation according to claim 8, characterized in that, The stem cells include mesenchymal stem cells and / or induced pluripotent stem cells, which are disposed on the outside of the hollow fiber tube in the form of three-dimensional culture microspheres or adherent cells.

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