Airlift type culture and separation device for outer vesicles of cucurbita moschata

Abstract

The utility model discloses an airlift type muskmelon outer vesicle culture and separation device which comprises a culture tank, the culture tank is a glass tank body with a big-end-up caliber, the top of the large caliber of the culture tank is connected with an exhaust valve through threads, and one side of the small caliber of the culture tank is fixedly connected with an air filter. One side of the air filter is fixedly connected with an air compressor, the culture tank is in threaded connection with a regulating valve, the output end of the regulating valve is connected with a pressure pump through a connecting pipe, the output end of the pressure pump is connected with an ultrafilter through a connecting pipe, and the bottom of the ultrafilter is provided with a discharge port; the outer surface of the side wall of the culture tank is in threaded connection with a temperature controller and a cooling water inlet, and the outer wall of the other side of the culture tank is in threaded connection with a cooling water outlet. The device is convenient to operate, fresh plant tissues do not need to be crushed traditionally, and the outer vesicles of the cucurbita moschata are cultured and enriched through airlift illumination suspension, so that the time and the labor cost are saved.

Description

Technical Field

[0001] This utility model relates to the field of plant cell culture and separation technology, and in particular to an airlift-type exovesicle culture and separation device for *Gnaphalium affine*. Background Technology

[0002] Bitter melon, also known as large bitter melon or green melon, has the effects of relieving summer heat, lowering blood sugar and blood lipids, and moisturizing the lungs and relieving cough. In recent years, studies have found that the fruit, stem, and root of bitter melon contain a variety of bioactive substances, which have anti-cancer and cancer-preventive effects and enhance the body's immunity. However, its production and clinical application are limited by the low content, low solubility, and low bioavailability of its medicinal components.

[0003] Plant exovesicles, as nanoscale vesicles secreted by plant cells, contain proteins, lipids, and nucleic acids. They possess a double-membrane structure, excellent membrane permeability, and strong ability to penetrate biological barriers, demonstrating potential applications in disease treatment. Studies have shown that plant exovesicles have natural targeting capabilities, delivering bioactive substances to specific sites, thereby improving drug utilization. For example, exosomes extracted from purslane have been found to have the potential to alleviate acute colitis. Ginseng-derived exosome nanovesicles have been found to serve as a highly efficient and safe carrier, efficiently delivering their contained active miRNAs to stem cells and inducing their differentiation into nerve cells, showing good effects in accelerating nerve regeneration and restoring conduction function.

[0004] Currently, plant exovesicles are mainly obtained by disrupting fresh plant tissue and using ultra-high-speed centrifugation. This method is time-consuming, labor-intensive, and suffers from low purity, inefficiency, and insufficient protection of exosome structures, making it difficult to scale up production. Therefore, there is an urgent need to develop an airlift-type exovesicle culture and separation device from *Trichosanthes kirilowii*. Utility Model Content

[0005] The purpose of this invention is to address the shortcomings of existing technologies by proposing an air-lift type exocarp culture and separation device for *Solanum tuberosum*.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] An airlift-type exovesicle culture and separation device for *Trichosanthes kirilowii* includes a culture tank, which is a glass tank with a larger diameter at the top and a smaller diameter at the bottom. The top of the culture tank with a larger diameter is connected to an exhaust valve via a threaded connection, and an air filter is fixedly connected to one side of the culture tank with a smaller diameter. An air compressor is fixedly connected to one side of the air filter. The culture tank is connected to a regulating valve via a threaded connection, and the output end of the regulating valve is connected to a pressure pump via a connecting pipe. The output end of the pressure pump is connected to an ultrafilter via a connecting pipe, and the bottom of the ultrafilter has a discharge port.

[0008] As a further embodiment of this utility model: a temperature controller and a cooling water inlet are connected to the outer surface of the side wall of the culture tank by a thread, and a cooling water outlet is connected to the outer wall of the other side of the culture tank by a thread.

[0009] As a further improvement of this utility model: the top of the culture tank is connected to a material tank via a pulsed injection pump through a threaded connection;

[0010] Add the callus stem cells and their culture medium to the material tank, and inject them into the culture tank by starting the pulsed perfusion pump. The temperature of the culture tank is adjusted by the temperature controller.

[0011] As a further improvement of this utility model: a pressure gauge, a pH electrode, and a dissolved oxygen electrode are fixedly connected to the top of the culture tank;

[0012] The air compressor is started to filter air through an air filter and then inject it from the bottom of the culture tank to form an airlift suspension culture. During this process, pH and dissolved oxygen electrodes monitor the dissolved oxygen concentration and pH value of the solution in the culture tank.

[0013] As a further improvement of this invention: the end of the ultrafilter is connected to one side of the material tank via a connecting pipe.

[0014] As a further improvement of this utility model, the two ends of the pulsating infusion pump are respectively connected to the top of the culture tank and one side of the material tank through conduits.

[0015] As a further improvement of this invention: the air compressor and air filter are connected to the small-diameter end of the culture tank by a thread.

[0016] Compared with the prior art, this utility model provides an airlift-type exovesicle culture and separation device for *Solanum tuberosum*, which has the following beneficial effects:

[0017] This invention relates to an airlift-type exovesicle culture and separation device for *Trichosanthes kirilowii*. This device is easy to operate and does not require traditional methods of crushing fresh plant tissue. It enriches exovesicles of *Trichosanthes kirilowii* through airlift-type light-induced suspension culture, saving time and labor costs.

[0018] This invention relates to an airlift-type exovesicle culture and separation device for Scutellaria baicalensis, which uses an airlift fermentation method to avoid the damage to Scutellaria baicalensis callus tissue and hairy root stem cells caused by the agitator of a stirred bioreactor, thus preventing a decrease in transformation yield.

[0019] This airlift-type *Solanum tuberosum* extravesicle culture and separation device effectively reduces production costs by separating *Solanum tuberosum* extravesicles and recovering *Solanum tuberosum* stem cells through ultrafiltration.

[0020] This invention relates to an airlift-type exovesicle culture and separation device for *Solanum tuberosum*. The separation of exovesicles from *Solanum tuberosum* is simple to operate, not easily contaminated, and suitable for medium-scale production.

[0021] The parts of this device not covered herein are the same as or can be implemented using existing technologies. This utility model has a simple structure and is easy to operate. Attached Figure Description

[0022] Figure 1 This is a schematic diagram of the structure of an airlift-type exovesicle culture and separation device for *Trichosanthes kirilowii* proposed in this utility model;

[0023] Figure 2 This is a schematic diagram of the airlift circulation of an airlift-type vesicle culture and separation device for *Solanum tuberosum* proposed in this utility model.

[0024] In the diagram: 1. Temperature controller; 2. Cooling water inlet; 3. Regulating valve; 4. Pressure pump; 5. Ultrafilter; 6. Discharge port; 7. Air compressor; 8. Air filter; 9. Culture tank; 10. Cooling water outlet; 11. Exhaust valve; 12. Material tank; 13. Pulsating injection pump; 14. Pressure gauge; 15. pH electrode; 16. Dissolved oxygen electrode. Detailed Implementation

[0025] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present utility model. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments.

[0026] In the description of this utility model, it should be understood that the terms "center", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this utility model and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this utility model.

[0027] In the description of this utility model, it should be noted that, unless otherwise explicitly specified and limited, the terms "installation," "connection," "linking," and "setting" should be interpreted broadly. For example, they can refer to a fixed connection or setting, a detachable connection or setting, or an integral connection or setting. Those skilled in the art can understand the specific meaning of the above terms in this utility model according to the specific circumstances.

[0028] An air-lift type exovesicle culture and separation device for *Solanum tuberosum*, such as Figure 1 and Figure 2As shown, the system includes a culture tank 9, which is a glass tank with a larger opening at the top and a smaller opening at the bottom. The top of the culture tank 9 with a larger opening is connected to an exhaust valve 11 via a thread, and an air filter 8 is fixedly connected to the side of the culture tank 9 with a smaller opening. An air compressor 7 is fixedly connected to the side of the air filter 8. The culture tank 9 is connected to a regulating valve 3 via a thread, and the output end of the regulating valve 3 is connected to a pressure pump 4 via a connecting pipe. The output end of the pressure pump 4 is connected to an ultrafilter 5 via a connecting pipe. The bottom of the ultrafilter 5 has a discharge port 6. The air compressor 7 and the air filter 8 are connected to the side of the culture tank 9 with a smaller opening via a thread.

[0029] The air compressor 7 is an Atlas Copco GA 11-30 VSD+, the air filter 8 is a Camfil FARR 30 / 30, the regulating valve 3 is a Swagelok SS-4BK-V51-1C, the pressure pump 4 is an Ismatec IPC-N 4, and the ultrafilter 5 is a Sartorius Vivaspin Turbo 15.

[0030] The outer surface of the side wall of the culture tank 9 is connected to a temperature controller 1 and a cooling water inlet 2 by threads. The outer wall of the other side of the culture tank 9 is connected to a cooling water outlet 10 by threads. The top of the culture tank 9 is connected to a material tank 12 by a pulsed filling pump 13 by threads. The temperature controller 1 is a Julabo FP80 and the pulsed filling pump 13 is a Masterflex L / S 7523-70.

[0031] The top of the culture tank 9 is fixedly connected to a pressure gauge 14, a pH electrode 15, and a dissolved oxygen electrode 16, and the signals of the pressure gauge 14, pH electrode 15, and dissolved oxygen electrode 16 are WIKA A-10, PH 10 / 20, and LDO101, respectively.

[0032] The end of the ultrafilter 5 is connected to one side of the material tank 12 via a connecting pipe, and the two ends of the pulsed perfusion pump 13 are respectively connected to the top of the culture tank 9 and one side of the material tank 12 via conduits.

[0033] In summary, the overall device eliminates the need for traditional fresh plant tissue pulverization. It enriches the exovesicles of *Trichosanthes kirilowii* through airlift-based light-induced suspension culture, saving time and labor costs. Furthermore, the airlift fermentation method avoids the damage to callus tissue and hairy root stem cells caused by the stirring paddles of stirred bioreactors, which would otherwise reduce the conversion yield. At the same time, the separation of exovesicles and the recovery of stem cells through ultrafiltration effectively reduces production costs. The separation of exovesicles is simple, less prone to bacterial contamination, and suitable for medium-scale production.

[0034] Working principle: Trichosanthes kirilowii callus stem cells and their culture medium are added to the material tank 12, and injected into the culture tank 9 by starting the pulsed perfusion pump 13. The temperature of the culture tank 9 is regulated by the thermostat 1. During this period, the air compressor 7 is started to inject air from the bottom of the culture tank 9 after being filtered by the air filter 8, forming an airlift suspension culture. During this period, the pH electrode 15 and the dissolved oxygen electrode 16 monitor the dissolved oxygen concentration and pH value of the solution in the culture tank 9. Subsequently, the gas generated by fermentation during the culture process is discharged from the culture tank 9 through the exhaust valve 3. The Trichosanthes kirilowii callus stem cells are separated from the Trichosanthes kirilowii outer vesicles by the ultrafilter 5. The separated stem cells are recycled back to the material tank 12.

[0035] The above description is only a preferred embodiment of the present utility model, but the protection scope of the present utility model is not limited thereto. Any equivalent substitutions or changes made by those skilled in the art within the technical scope disclosed in the present utility model, based on the technical solution and the inventive concept of the present utility model, should be included within the protection scope of the present utility model.

Claims

1. An airlift-type exovesicle culture and separation device for *Gnaphalium affine*, comprising a culture vessel (9), wherein the culture vessel (9) is a glass jar with a larger opening at the top and a smaller opening at the bottom, characterized in that, The culture tank (9) has an exhaust valve (11) connected to the top of the large diameter via a threaded connection, and an air filter (8) is fixedly connected to the side of the small diameter of the culture tank (9). An air compressor (7) is fixedly connected to the side of the air filter (8). The culture tank (9) has a regulating valve (3) connected via a threaded connection. The output end of the regulating valve (3) is connected to a pressure pump (4) via a connecting pipe. The output end of the pressure pump (4) is connected to an ultrafilter (5) via a connecting pipe. The bottom of the ultrafilter (5) has a discharge port (6).

2. The airlift-type exovesicle culture and separation device for *Solanum tuberosum* according to claim 1, characterized in that, The outer surface of the side wall of the culture tank (9) is connected to a temperature controller (1) and a cooling water inlet (2) by thread, and the outer wall of the other side of the culture tank (9) is connected to a cooling water outlet (10) by thread.

3. The airlift-type exovesicle culture and separation device for *Solanum tuberosum* according to claim 2, characterized in that, The top of the culture tank (9) is connected to a material tank (12) via a pulsating injection pump (13) through a threaded connection.

4. The airlift-type exovesicle culture and separation device for *Solanum tuberosum* according to claim 3, characterized in that, A pressure gauge (14), a pH electrode (15), and a dissolved oxygen electrode (16) are fixedly connected to the top of the culture tank (9).

5. The airlift-type exovesicle culture and separation device for *Solanum tuberosum* according to claim 1, characterized in that, The end of the ultrafilter (5) is connected to one side of the material tank (12) via a connecting pipe.

6. The airlift-type exovesicle culture and separation device for *Solanum tuberosum* according to claim 3, characterized in that, The two ends of the pulsating infusion pump (13) are connected to the top of the culture tank (9) and one side of the material tank (12) respectively through conduits.

7. The airlift-type exovesicle culture and separation device for *Solanum tuberosum* according to claim 1, characterized in that, The air compressor (7) and air filter (8) are connected to the small-diameter end of the culture tank (9) by threads.

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