Stem cell exosome biological agent preparation system and method

Abstract

The stem cell exosome biological preparation preparation system comprises an isolation bin, a centrifugal assembly, a filtering assembly, a biological culture assembly, a finished product storage assembly and an intelligent transfer module, through the collaborative design of the totally-closed isolation bin and the intelligent transfer module, the pollution risk and the efficiency bottleneck caused by manual operation in the traditional preparation process are effectively solved; an automatic pipeline embedded in the isolation bin and an infusion pump achieve whole-process closed transmission of liquid, and the risk of cross contamination is thoroughly eliminated; the multi-stage series transfer warehouse is integrated with a high-precision sensor, key indexes such as purity and activity of the exosome can be detected online in real time, a control module is linked to automatically screen qualified products, the preparation period is greatly shortened, and the exosome yield and batch-to-batch consistency are remarkably improved. Meanwhile, a waste liquid recovery and intelligent cleaning mechanism is arranged in the system, reagent loss and manual maintenance requirements are reduced, and efficient, stable and safe technical support is provided for large-scale production of clinical stem cell exosome preparations.

Description

Technical Field

[0001] The present invention relates to the field of biological treatment, and in particular to a system and method for preparing an extracellular vesicle biological preparation. Background Art

[0002] As an important carrier of regenerative medicine and precision treatment, stem cell exosome biopharmaceuticals have been widely used in the fields of tissue repair, immune regulation (such as the treatment of autoimmune diseases) and drug delivery. Its preparation process mainly includes stem cell expansion (such as high-density culture of mesenchymal stem cells in a bioreactor), exosome separation (through ultracentrifugation, size exclusion chromatography or ultrafiltration technology) and purification (removal of impurities such as free proteins and cell debris). In the existing equipment system, bioreactors are used to maintain the activity and secretory function of stem cells, centrifuges (such as differential centrifugation equipment) are used to remove cell debris and enrich exosomes in stages, and ultrafiltration devices achieve exosome concentration through molecular weight retention. Although these devices have basic functions in a single link, their decentralized design makes the connection between each link dependent on manual intervention, and lacks a unified intelligent control module, making it difficult to meet the efficiency and consistency requirements of large-scale production.

[0003] In the current preparation process, sample transfer between multiple devices (such as transferring liquid into an ultrafiltration device after centrifugation and transferring exosomes into a freeze-dryer after purification) is highly dependent on manual operations and presents significant bottlenecks: First, frequent opening of the lid and manual pipetting increase the risk of cross-contamination, especially when the exosome concentration is extremely low, where trace contamination can significantly affect the safety of the preparation; second, manual transfer efficiency is low, and single-batch preparation requires multiple interruptions, extending the overall cycle to more than 24 hours; third, the detection of key parameters (such as exosome concentration and particle size distribution) requires offline sampling and the use of independent instruments (such as nanoparticle tracking analyzers), resulting in delayed data feedback and inability to guide process adjustments in real time, leading to quality fluctuations between batches. These problems have seriously restricted the clinical translation and industrial production of stem cell exosome preparations. Summary of the Invention

[0004] In order to solve the above problems, the present invention discloses the following technical solutions:

[0005] A stem cell exosome biological preparation system, characterized by comprising:

[0006] The isolation chamber comprises a chamber body and a main control panel provided on the surface of the chamber body;

[0007] The centrifugal assembly is located in the chamber and can centrifuge the fine liquid.

[0008] A filtration component, which is used to filter the cell fluid after centrifugation;

[0009] A biological culture assembly for receiving the liquid filtered by the filtering assembly and carrying out biological culture;

[0010] A finished product storage assembly for receiving the liquid cultured by the biological culture assembly;

[0011] An intelligent transfer module comprising a transfer chamber communicated with the centrifugal assembly, the filtering assembly, the biological culture assembly and the finished product storage assembly through pipes, and a detection sensor arranged in the transfer chamber, wherein the products of the centrifugal assembly, the filtering assembly and the biological culture assembly all flow into the transfer chamber first and then enter the next level of processing equipment from the transfer chamber;

[0012] The surfaces of the pipes are all provided with liquid pumping pumps capable of pumping liquid.

[0013] On the basis of the above technical solution, the application can be further improved as follows.

[0014] Further, the number of the transfer chambers is at least three, and all the transfer chambers are communicated in sequence through pipes, and electronic valves are arranged on the pipes.

[0015] Further, a waste liquid collecting assembly arranged in the interior of the chamber body is further included, and the waste liquid collecting assembly is communicated with the transfer chamber and the biological culture assembly.

[0016] Further, a primary liquid chamber arranged in the isolation chamber is further included, and the primary liquid chamber is communicated with the liquid inlet end of the centrifugal assembly.

[0017] Further, a cleaning assembly is further included, and the cleaning assembly comprises a liquid storage member arranged in the interior of the isolation chamber and a spray head communicated with the liquid storage member.

[0018] Further, the liquid storage member is communicated with the primary liquid chamber, the centrifugal assembly, the filtering assembly and the biological culture assembly through pipes, and the spray head is arranged in the interior of the primary liquid chamber, the centrifugal assembly, the filtering assembly and the biological culture assembly.

[0019] Further, the surface of the transfer chamber is provided with an observation window for an operator to observe the interior thereof.

[0020] Further, a hot air drying unit is further included, and the hot air drying unit is communicated with the primary liquid chamber, the centrifugal assembly, the filtering assembly and the biological culture assembly through a gas conveying pipe, and the surface of the gas conveying pipe is provided with an electronic gas valve for closing the gas outlet thereof.

[0021] A stem cell exosome biological preparation preparation method, which adopts the stem cell exosome biological preparation preparation system of claims 1-8, and comprises the following steps:

[0022] S1, inject stem cell culture liquid or tissue extraction liquid into the primary liquid chamber in the isolation chamber, and start the liquid pumping pump of the main control panel to convey the liquid in the primary liquid chamber to the liquid inlet end of the centrifugal assembly through the pipes;

[0023] S2, the centrifugation component runs in stages according to the preset program to remove cell debris and enrich exosomes;

[0024] After centrifugation, the liquid pump draws the centrifuged liquid into the first transfer chamber, and the detection sensor detects the exosome concentration in real time;

[0025] S3. If the test meets the standard, the electronic valve of the first transfer chamber opens, and the liquid enters the filter assembly through the pipeline

[0026] S4: The filtered liquid flows into the second transfer tank, and the detection sensor verifies the purity for the second time. If it is qualified, the next process is triggered;

[0027] S5. The filtered qualified liquid is transported from the second transfer chamber to the biological culture component, and the serum-free culture medium circulation and dissolved oxygen control are started;

[0028] S6. After the culture is completed, the liquid is tested for exosome activity in the third transfer chamber and then transported to the finished product storage component by a liquid pump;

[0029] S7, the waste liquid in the transfer chamber and the biological culture component switches the path through the electronic valve and flows into the waste liquid collection component;

[0030] S8. The cleaning component is started, and the cleaning liquid is pumped into each component to clean each processing unit. After the cleaning is completed, the waste liquid enters the waste liquid collection component.

[0031] Furthermore, in step S4, if the test fails, the liquid is transferred to a waste liquid collection component.

[0032] Beneficial effects

[0033] Compared to existing technologies, this invention effectively addresses the contamination risks and efficiency bottlenecks caused by manual operations during the traditional preparation process through the collaborative design of a fully enclosed isolation chamber and an intelligent transfer module. Automated piping and pumps embedded in the isolation chamber ensure fully enclosed liquid transfer, completely eliminating the risk of cross-contamination. The multi-stage serial transfer chamber integrates high-precision sensors for real-time online monitoring of key exosome indicators such as purity and activity, and automatically selects qualified products through a control module, significantly shortening the preparation cycle and significantly improving exosome yield and batch-to-batch consistency. Furthermore, the system incorporates built-in waste liquid recovery and intelligent cleaning mechanisms to reduce reagent consumption and manual maintenance requirements, providing efficient, stable, and safe technical support for the large-scale production of clinical-grade stem cell exosome preparations. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0035] Figure 1 This is a schematic diagram of the invention's front cross-sectional structure;

[0036] Figure 2 It is a structural diagram of the invention;

[0037] In the accompanying drawings, the components represented by the reference numerals are as follows:

[0038] 1. Isolation chamber; 11. Chamber body; 12. Main control panel; 2. Centrifugal assembly; 3. Filtration assembly; 4. Biological culture assembly; 5. Finished product storage assembly; 6. Intelligent transfer module; 7. Waste liquid collection assembly; 8. Initial liquid chamber; 9. Cleaning assembly. DETAILED DESCRIPTION

[0039] To facilitate understanding of the present invention, the present invention will be described more fully below with reference to the accompanying drawings. The drawings illustrate several embodiments of the present invention. However, the present invention may be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a more thorough and comprehensive understanding of the present invention.

[0040] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention pertains. The terms used in this specification of the present invention are for the purpose of describing specific embodiments only and are not intended to limit the present invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0041] It should be noted that similar reference numerals and letters denote similar items in the following drawings, and therefore, once an item is defined in one drawing, it does not need to be further defined or explained in subsequent drawings.

[0042] It should be noted that the terms "center", "up", "down", "left", "right", "vertical", "horizontal", "inside", "outside", etc., indicating the orientation or position relationship, are based on the orientation or position relationship shown in the accompanying drawings, or are the orientation or position relationship in which the technical product is usually placed when in use. They are only for the convenience of describing this technology and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation. Therefore, they should not be understood as a limitation on this technology.

[0043] Furthermore, terms such as "horizontal," "vertical," and "overhanging" do not necessarily imply that a component must be absolutely horizontal or overhanging, but rather that it can be slightly tilted. For example, "horizontal" simply means that its direction is more horizontal than "vertical," and does not mean that the structure must be completely horizontal, but rather that it can be slightly tilted.

[0044] See also Figure 1 The stem cell exosome biological preparation system includes an isolation chamber 1, a centrifugal component 2, a filtration component 3, a biological culture component 4, a finished product storage component 5, and an intelligent transfer module 6.

[0045] Among them, the isolation chamber 1 is a common box structure in the prior art, including a chamber body 11 and a main control panel 12 arranged on the surface of the chamber body 11. The surface of the chamber body 11 may also be provided with an opening and closing door and a ventilation device.

[0046] The centrifugal assembly 2 is arranged in the chamber 11 and can centrifuge the fine liquid.

[0047] The filter assembly 3 is used to filter the cell fluid after centrifugation;

[0048] The biological culture component 4 is used to receive the liquid filtered by the filtering component 3 and perform biological culture;

[0049] The finished product storage component 5 is used to receive the liquid cultured by the biological culture component 4;

[0050] The centrifugal component 2, the filtration component 3, and the biological culture component 4 are all commonly used devices for cultivating stem cell exosomes in the prior art.

[0051] The intelligent transfer module 6 includes a transfer warehouse connected to the centrifugal component 2, the filtration component 3, the biological culture component 4 and the finished product storage component 5 through a pipeline, and a detection sensor arranged in the transfer warehouse. The products of the centrifugal component 2, the filtration component 3 and the biological culture component 4 first flow into the transfer warehouse, and then enter the next-level processing equipment from the transfer warehouse, so as to temporarily store the intermediate products and realize process connection, avoid manual transfer contamination, and at the same time, ensure that the intermediate products meet the standards through the detection sensors. A pipeline loop can also be set up. When the reactants do not meet the standards, the system chooses whether to discard the intermediate products or re-enter the upper-level equipment. The detection sensors can be nanoparticle tracking analysis sensors, electrochemical sensors, and temperature sensors, which are designed to detect different properties of the intermediate products in the transfer warehouse.

[0052] The surface of the pipeline is provided with a liquid pump capable of pumping liquid. The liquid pump can be driven by a high-precision stepping motor and can adaptively adjust the suction rate according to the viscosity of the liquid.

[0053] The number of the transfer warehouses is at least three, and all the transfer warehouses are connected in sequence through pipelines. Electronic valves are provided on the pipelines. Multi-stage transfer warehouses are connected in series to realize staged detection (concentration → purity → activity). The electronic valves automatically divert qualified products and waste liquid, effectively improving the yield of finished products.

[0054] Example 1

[0055] In order to avoid environmental pollution caused by waste liquid discharge, the present application also includes a waste liquid collection component 7 arranged inside the warehouse body 11, and the waste liquid collection component 7 is connected to the transfer warehouse and the biological culture component 4. It also includes an initial liquid warehouse 8 arranged in the isolation warehouse 1, and the initial liquid warehouse 8 is connected to the liquid inlet end of the centrifugal component 2, standardizing the initial sample loading process and reducing the complexity of the operation.

[0056] Example 2

[0057] Different from Example 1, it also includes a cleaning component 9, which includes a liquid storage part located inside the isolation chamber 1 and a spray head connected to the liquid storage part. The liquid storage part can store a variety of cleaning agents to meet different sterilization requirements. The liquid storage part is connected to the initial liquid chamber 8, the centrifugal component 2, the filter component 3, and the biological culture component 4 through a pipeline. The spray head is located inside the initial liquid chamber 8, the centrifugal component 2, the filter component 3, and the biological culture component 4. The spray head can be a pulse spray cleaning liquid to thoroughly remove the residues in the pipeline and components. Through the design of the pipeline, the entire system can be cleaned in situ to avoid the risk of disassembly contamination. At the same time, the surface of the transfer chamber is provided with an observation window for the operator to observe its interior, which also allows the operator to observe the intermediate product.

[0058] Example 3

[0059] Different from Example 2, the present application also includes a hot air drying unit, which is connected to the initial liquid tank 8, centrifugal component 2, filter component 3, and biological culture component 4 through an air pipe, and an electronic air valve is provided on the surface of the air pipe for closing its air outlet.

[0060] To explain the above technical solution in detail, a method for preparing a stem cell exosome biopharmaceutical is now disclosed, which uses a stem cell exosome biopharmaceutical preparation system and includes the following steps:

[0061] S1. Inject stem cell culture fluid or tissue extract into the initial liquid tank 8 in the isolation chamber 1. The main control panel 12 starts the liquid pump to transport the liquid in the initial liquid tank 8 to the liquid inlet end of the centrifugal assembly 2 through the pipeline.

[0062] S2, centrifugation component 2 runs in stages according to the preset program to remove cell debris and enrich exosomes;

[0063] After centrifugation, the liquid pump draws the centrifuged liquid into the first transfer chamber, and the detection sensor detects the exosome concentration in real time;

[0064] S3. If the test meets the standard, the electronic valve of the first transfer chamber opens, and the liquid enters the filter component 3 through the pipeline.

[0065] S4: The filtered liquid flows into the second transfer chamber. If the purity is qualified, the next process is triggered. If it fails the test, the liquid is transferred to the waste liquid collection component 7.

[0066] S5: The filtered qualified liquid is transported from the second transfer chamber to the biological culture component 4, and the serum-free culture medium circulation and dissolved oxygen control are started;

[0067] S6. After the culture is completed, the liquid is tested for exosome activity in the third transfer chamber and then transported to the finished product storage component 5 by the liquid pump;

[0068] S7, the waste liquid in the transfer chamber and the biological culture component 4 switches the path through the electronic valve and flows into the waste liquid collection component 7;

[0069] S8, the cleaning component 9 is started, and the cleaning liquid is pumped into each component to clean each processing unit. After the cleaning is completed, the waste liquid enters the waste liquid collection component 7.

[0070] In the description of this technology, it should also be noted that, unless otherwise specified or limited, the terms "disposed," "installed," "connected," and "connected" should be understood broadly. For example, they can refer to fixed, detachable, or integral connections; mechanical or electrical connections; direct or indirect connections through an intermediate medium; and internal communication between two components. Those skilled in the art will understand the specific meanings of these terms in this technology based on the specific circumstances.

[0071] The above description is only a preferred specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any technician familiar with the technical field, within the technical scope disclosed by the present invention, who makes equivalent replacements or changes based on the technical solution and inventive concept of the present invention, should be covered by the scope of protection of the present invention.

Claims

1. A stem cell exosome biological preparation system, characterized in that: include: An isolation chamber (1) comprises a chamber body (11) and a main control panel (12) disposed on a surface of the chamber body (11); The centrifugal assembly (2) is arranged in the chamber (11) and is capable of centrifuging the fine liquid. A filtering component (3), which is used to filter the cell fluid after centrifugation; A biological culture component (4) is used to receive the liquid filtered by the filtering component (3) and perform biological culture; A finished product storage component (5) is used to receive the liquid cultured by the biological culture component (4); An intelligent transfer module (6) includes a transfer chamber connected to a centrifugal assembly (2), a filtration assembly (3), a biological culture assembly (4), and a finished product storage assembly (5) through a pipeline, and a detection sensor disposed in the transfer chamber. Products of the centrifugal assembly (2), the filtration assembly (3), and the biological culture assembly (4) first flow into the transfer chamber and then enter the next-level processing equipment from the transfer chamber. The surfaces of the pipelines are all provided with liquid pumps capable of pumping liquid.

2. A stem cell exosome biological preparation system according to claim 1, characterized in that: The number of the transfer chambers is at least three, and all the transfer chambers are connected in sequence through pipelines, and electronic valves are provided on the pipelines.

3. A stem cell exosome biological preparation system according to claim 1, characterized in that: It also includes a waste liquid collection component (7) arranged inside the warehouse body (11), and the waste liquid collection component (7) is connected to the transfer warehouse and the biological culture component (4).

4. A stem cell exosome biological preparation system according to claim 1, characterized in that: It also includes a primary liquid tank (8) arranged in the isolation tank (1), and the primary liquid tank (8) is communicated with the liquid inlet end of the centrifugal component (2).

5. The stem cell exosome biological preparation system according to claim 1, characterized in that: It also includes a cleaning component (9), which includes a liquid storage component arranged inside the isolation chamber (1) and a spray head connected to the liquid storage component.

6. A stem cell exosome biological preparation system according to claim 1, characterized in that: The liquid storage component is connected to the initial liquid tank (8), the centrifugal component (2), the filter component (3), and the biological culture component (4) through a pipeline, and the spray head is located inside the initial liquid tank (8), the centrifugal component (2), the filter component (3), and the biological culture component (4).

7. The stem cell exosome biological preparation system according to claim 1, characterized in that: The surface of the transfer bin is provided with an observation window for the operator to observe the interior thereof.

8. The stem cell exosome biological preparation system according to claim 1, characterized in that: The invention also comprises a hot air drying unit, which is connected to the initial liquid tank (8), the centrifugal component (2), the filter component (3), and the biological culture component (4) through an air supply pipe, and an electronic air valve for closing the air outlet of the air supply pipe is provided on the surface of the air supply pipe.

9. A method for preparing a stem cell exosome biopharmaceutical, comprising the steps of: S1. Inject stem cell culture fluid or tissue extract into the initial liquid chamber (8) in the isolation chamber (1). The main control panel (12) starts the liquid pump to transport the liquid in the initial liquid chamber (8) to the liquid inlet end of the centrifugal assembly (2) through the pipeline; S2, the centrifugation component (2) runs in stages according to a preset program to remove cell debris and enrich exosomes; After centrifugation, the liquid pump draws the centrifuged liquid into the first transfer chamber, and the detection sensor detects the exosome concentration in real time; S3. If the test meets the standard, the electronic valve of the first transfer chamber opens, and the liquid enters the filter assembly (3) through the pipeline S4: The filtered liquid flows into the second transfer tank, and the detection sensor verifies the purity for the second time. If it is qualified, the next process is triggered; S5, the filtered qualified liquid is transported from the second transfer chamber to the biological culture component (4), and the serum-free culture medium circulation and dissolved oxygen control are started; S6. After the culture is completed, the liquid is tested for exosome activity in the third transfer chamber and then transported to the finished product storage component (5) by a liquid pump; S7, the waste liquid in the transfer chamber and the biological culture component (4) switches the path through the electronic valve and flows into the waste liquid collection component (7); S8, the cleaning component (9) is started, and the cleaning liquid is pumped into each component to clean each processing unit. After the cleaning is completed, the waste liquid enters the waste liquid collection component (7).

10. The method for preparing a stem cell exosome biological preparation according to claim 9, characterized in that: In step S4, if the test fails, the liquid is transferred to the waste liquid collection component (7).

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