Culture equipment for promoting secretion of cell exosomes

Abstract

The utility model belongs to the technical field of bioengineering, and particularly relates to culture equipment for promoting cell exosome secretion, which comprises a constant-temperature box, a culture table is slidably connected in the constant-temperature box, a vertical plate is fixedly connected on the culture table, the surface of the vertical plate is rotatably connected with a U-shaped support through a shaft, and the U-shaped support is fixedly connected with the culture table. The interior of the support is detachably connected with a reactor through a connecting assembly, a culture assembly is installed on the reactor, the culture assembly comprises a plurality of conducting strips and a threaded connecting end, and the conducting strips are fixedly installed at the bottom of the reactor; according to the utility model, the gas suction pump or the gas output pump is operated to pressurize or depressurize the interior of the reactor so as to change the internal pressure of the reactor, and when the gas pressure sensor detects that the gas pressure meets the requirement, the electrical stimulation device is operated to enable the conducting strip to generate micro current so as to stimulate cells in a culture solution to generate exosomes.

Description

Technical Field

[0001] The utility model relates to the technical field of bioengineering, in particular to a culture device for promoting the secretion of extracellular vesicles. Background Art

[0002] Extracellular vesicles are bioactive substances naturally secreted by cells, composed of lipids, proteins, nucleic acids, etc., and are vesicles with a diameter of 30 - 150 nm. As an important way for stem cells to exert biological effects, they have the advantages of low immunogenicity, high stability, nanoscale ability to cross the blood-brain barrier, high activity, and low tumorigenicity compared with stem cells. Through various mechanisms such as binding to target cells, endocytosis, and membrane fusion, the expression information of donor cells is transmitted to recipient cells, enabling the reprogramming of target cells to exert biological regulatory functions. It has now been confirmed that extracellular vesicles have mechanism characteristics such as regulating inflammatory responses, promoting cell proliferation and migration, promoting angiogenesis, and regulating matrix reconstruction.

[0003] A bioreactor is the core equipment for large-scale suspension culture of cells, which can effectively increase the culture density of cells per unit volume. Animal cell culture bioreactors are divided into stirred cell culture bioreactors and non-stirred cell culture bioreactors according to whether there is a stirring paddle inside. The stirred cell culture reactor generates eddies through the rotation of the stirring paddle to complete the aeration and oxygen supply of the culture medium, but the mechanical shear force generated by this method is relatively large and is extremely likely to damage cells.

[0004] The existing Chinese patent with the publication number CN219824203U discloses a culture device for promoting the secretion of extracellular vesicles, which includes a cell culture device, a gas transmission device, an electric stimulation device, a filtration device, and a control device. The cell culture device contains a culture medium, the gas transmission device is used to introduce gas into or extract gas from the cell culture device to change the internal pressure of the bottle of the cell culture device, the electric stimulation device applies an electric current to the culture medium through a cable to stimulate the cells in the culture medium to produce extracellular vesicles, the filtration device is connected to the cell culture device to filter the culture medium, and the control device is electrically connected to the gas transmission device and the electric stimulation device respectively to control the start and stop of the two.

[0005] In view of the above and existing related technologies, the inventor believes that the following defects often exist: during the culture process of extracellular vesicles, the temperature is not well controlled and is maintained at room temperature for culture. However, the survival environment of cells in the human body is relatively constant and needs to be improved. Therefore, a culture device for promoting the secretion of extracellular vesicles is proposed to solve the above problems. Summary of the Utility Model

[0006] In order to make up for the deficiencies of the prior art and solve the above-mentioned technical problems, the utility model proposes a culture device for promoting the secretion of extracellular vesicles.

[0007] The technical solution adopted by the present utility model to solve its technical problems is as follows: A culture device for promoting the secretion of extracellular vesicles of the present utility model includes a thermostatic chamber, inside which a culture table is slidably connected. A vertical plate is fixedly connected to the culture table. The surface of the vertical plate is rotatably connected with a U-shaped bracket through a shaft. A reactor is detachably connected inside the bracket through a connection component. A culture component is installed on the reactor. The culture component includes a plurality of conductive sheets and threaded connection ends. The conductive sheets are fixedly installed at the bottom of the reactor. The threaded connection ends are fixedly installed on the lower side wall of the reactor. A rotary joint is threadedly connected inside the threaded connection end. The rotary joint is fixedly connected to a three-way hose. Above the culture table, a gas suction pump and a gas output pump are fixedly connected. Both ends of the gas suction pump and the gas output pump are fixedly connected to both ends of the three-way hose respectively. Solenoid valves are fixedly connected to both ends of the three-way hose respectively. When the solenoid valves are closed, the reactor remains sealed. When the gas suction pump or the gas output pump operates, the solenoid valve on this side is synchronously opened, so that the gas suction pump or the gas output pump is connected to the reactor through the three-way hose. An electric stimulation device is fixedly connected to the culture table. The electric stimulation device is electrically connected to the conductive sheets. A culture solution is placed in the reactor. By operating the gas suction pump or the gas output pump, the internal pressure of the reactor is increased or decreased to change the internal pressure of the reactor. When the air pressure sensor detects that the gas pressure meets the requirements, the electric stimulation device is operated to generate a microcurrent on the conductive sheets to stimulate the cells in the culture solution to produce extracellular vesicles.

[0008] Preferably, the culture component further includes a current induction device. The current induction device is fixedly installed at the bottom of the reactor and is electrically connected to the electric stimulation device. The magnitude of the microcurrent received by the cells can be measured through the current induction device, avoiding excessive microcurrent from damaging the cells and also avoiding too small microcurrent from failing to achieve an effective stimulation effect.

[0009] Preferably, a detection interface is installed on the surface of the rotary joint, and a pressure sensor is fixedly connected to the detection interface to detect the air pressure inside the reactor through the pressure sensor.

[0010] Preferably, the reactor includes a storage chamber, a filtration chamber, and a reaction chamber from top to bottom, and the storage chamber, the filtration chamber, and the reaction chamber are sequentially threadedly connected. The reactor is formed by combining the storage chamber, the filtration chamber, and the reaction chamber. After use, it is convenient to disassemble and clean.

[0011] Preferably, a filter layer is fixedly connected inside the filtration chamber.

[0012] Preferably, the filter layer adopts a hollow fiber filter membrane.

[0013] Preferably, the connection component includes a fixed plug and a movable plug. The fixed plug is fixedly installed at one end of the bracket, and the movable plug penetrates and is slidably installed at the other end of the bracket. When installing the reactor, first insert the fixed plug into the socket, and then the movable plug is inserted into the socket on the other side by the elastic force of the spring, thereby fixing the reactor on the bracket. By installing the reactor in the incubator to control the temperature during cultivation, the cells are kept at a suitable growth temperature. After the cell culture cycle ends, the reactor is inverted by rotating 180° through the driver, so that the culture solution is filtered and separated through the filter layer, thereby separating exosomes from other impurities such as organelles, and keeping the whole culture process at a constant temperature.

[0014] Preferably, a spring is sleeved on the surface of the movable plug.

[0015] Preferably, the connection component further includes two sockets, and the sockets are fixedly installed on both sides of the reactor.

[0016] Preferably, the connection component further includes a driver, and the driver is fixedly installed on the side wall of the vertical plate, and the driving end of the driver is fixedly connected to the shaft of the bracket.

[0017] The beneficial effects of the present utility model are as follows:

[0018] 1. By operating the gas inhalation pump or the gas output pump to increase or decrease the pressure inside the reactor, the internal pressure of the reactor is changed. When the gas pressure sensor detects that the gas pressure meets the requirements, the electric stimulation device is operated to generate a microcurrent on the conductive sheet to stimulate the cells in the culture solution to produce exosomes.

[0019] 2. By installing the reactor in the incubator to control the temperature during cultivation, the cells are kept at a suitable growth temperature. After the cell culture cycle ends, the reactor is inverted by rotating 180° through the driver, so that the culture solution is filtered and separated through the filter layer, thereby separating exosomes from other impurities such as organelles, and keeping the whole culture process at a constant temperature without manual operation. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the technical solutions in the embodiments of the present utility model or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the following drawings are only some embodiments of the present utility model. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0021] Figure 1 It is a schematic diagram of the overall structure of the present utility model;

[0022] Figure 2 Structural schematic diagram of the culture platform of the present utility model;

[0023] Figure 3 Structural schematic diagram of the culture component of the present utility model;

[0024] Figure 4 Cross-sectional view of the reactor of the present utility model;

[0025] Figure 5 Structural schematic diagram of the connection component of the present utility model.

[0026] In the figure: 1, constant temperature box; 2, culture platform; 3, vertical plate; 4, bracket; 5, reactor; 51, storage chamber; 52, filtration chamber; 53, reaction chamber; 54, filtration layer; 6, culture component; 61, conductive sheet; 62, current induction device; 63, threaded connection end; 64, rotary joint; 65, three-way hose; 66, air pressure sensor; 7, gas suction pump; 8, gas output pump; 9, solenoid valve; 10, electrical stimulation device; 11, connection component; 111, fixed plug; 112, movable plug; 113, spring; 114, socket; 115, driver. Specific embodiments

[0027] The technical solutions in the embodiments of the present utility model will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present utility model. Obviously, the described embodiments are only a part of the embodiments of the present utility model, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present utility model without making creative efforts shall fall within the protection scope of the present utility model.

[0028] Embodiment 1

[0029] Please refer to Figures 1-4As shown in the figure, a culture device for promoting the secretion of extracellular vesicles includes an incubator 1. Inside the incubator 1, there is a slidably connected culture table 2. On the culture table 2, there is a vertically fixed plate 3. The surface of the vertical plate 3 is rotatably connected with a U-shaped bracket 4 through a shaft. Inside the bracket 4, there is a reactor 5 detachably connected through a connection assembly 11. On the reactor 5, there is a culture assembly 6. The culture assembly 6 includes several conductive sheets 61 and threaded connection ends 63. The conductive sheets 61 are fixedly installed at the bottom of the reactor 5, and the threaded connection ends 63 are fixedly installed on the lower side wall of the reactor 5. Inside the threaded connection ends 63, there is a rotatable joint 64 threadedly connected. The rotatable joint 64 is fixedly connected with a three-way hose 65. Above the culture table 2, there are a gas suction pump 7 and a gas output pump 8 fixedly connected. The two ends of the gas suction pump 7 and the gas output pump 8 are respectively fixedly connected with the two ends of the three-way hose 65. The two ends of the three-way hose 65 are respectively fixedly connected with solenoid valves 9. When the solenoid valves 9 are closed, the reactor 5 remains sealed. When the gas suction pump 7 or the gas output pump 8 operates, the solenoid valve 9 on this side is synchronously opened, so that the gas suction pump 7 or the gas output pump 8 is connected to the reactor 5 through the three-way hose 65. On the culture table 2, there is an electrical stimulation device 10 fixedly connected. The electrical stimulation device 10 is electrically connected with the conductive sheets 61. The reactor 5 contains a culture solution. By operating the gas suction pump 7 or the gas output pump 8, the internal pressure of the reactor 5 is increased or decreased to change the internal pressure of the reactor 5. When the air pressure sensor 66 detects that the gas pressure meets the requirements, the electrical stimulation device 10 is operated to generate a microcurrent on the conductive sheets 61 to stimulate the cells in the culture solution to produce extracellular vesicles.

[0030] The culture assembly 6 further includes a current induction device 62. The current induction device 62 is fixedly installed at the bottom of the reactor 5, and the current induction device 62 is electrically connected with the electrical stimulation device 10. The magnitude of the microcurrent received by the cells can be measured through the current induction device 62, avoiding excessive microcurrent from damaging the cells and also avoiding too small microcurrent from failing to achieve an effective stimulation effect.

[0031] On the surface of the rotatable joint 64, there is a detection interface. A pressure sensor 66 is fixedly connected to the detection interface. The air pressure inside the reactor 5 is detected through the pressure sensor 66.

[0032] The reactor 5 includes a storage chamber 51, a filtration chamber 52, and a reaction chamber 53 from top to bottom, and the storage chamber 51, the filtration chamber 52, and the reaction chamber 53 are sequentially threadedly connected. The reactor 5 is formed by combining the storage chamber 51, the filtration chamber 52, and the reaction chamber 53. After use, it is convenient to disassemble and clean.

[0033] Inside the filtration chamber 52, there is a filter layer 54 fixedly connected.

[0034] The filter layer 54 uses a hollow fiber filter membrane.

[0035] Example 2

[0036] Comparing with the first comparative example, please refer to Figures 4-5 As shown, the present utility model provides another implementation manner. The connection assembly 11 includes a fixed plug 111 and a movable plug 112. The fixed plug 111 is fixedly installed at one end of the bracket 4, and the movable plug 112 penetrates and is slidably installed at the other end of the bracket 4. When installing the reactor 5, first insert the fixed plug 111 into the socket 114, and then the movable plug 112 is inserted into the socket 114 on the other side by the elastic force of the spring 113, thereby fixing the reactor 5 on the bracket 4. By installing the reactor 5 in the incubator 1 to control the temperature during cultivation, the cells are at a suitable growth temperature. After the cell culture cycle ends, the reactor 5 is inverted by rotating 180° through the driver 115, so that the culture solution is filtered and separated through the filter layer 54, thereby separating exosomes from other impurities such as organelles, making the whole cultivation process in a constant temperature state and without manual operation.

[0037] A spring 113 is sleeved on the surface of the movable plug 112.

[0038] The connection assembly 11 further includes two sockets 114, and the sockets 114 are fixedly installed on both sides of the reactor 5.

[0039] The connection assembly 11 further includes a driver 115. The driver 115 is fixedly installed on the side wall of the vertical plate 3, and the driving end of the driver 115 is fixedly connected to the shaft of the bracket 4.

[0040] Those skilled in the art connect all the electrical components in this case to their adapted power supplies through wires, and should select a suitable controller according to the actual situation and electrically connect it to devices such as the incubator 1, the current sensing device 62, the electrical stimulation device 10, the air pressure sensor 66, the gas inhalation pump 7, the gas output pump 8, the solenoid valve 9, and the driver 115 to meet the control requirements. For the specific connection and control sequence, reference should be made to the sequence of the electrical components working in sequence in the following working principle to complete the electrical connection. The detailed connection means are well-known techniques in the art. The following mainly introduces the working principle and process, and will not explain the electrical control.

[0041] Working principle: The reactor 5 contains a culture solution. By operating the gas inhalation pump 7 or the gas output pump 8 to increase or decrease the pressure inside the reactor 5 to change the internal pressure of the reactor 5. When the air pressure sensor 66 detects that the gas pressure meets the requirements, the electrical stimulation device 10 is operated to generate a microcurrent on the conductive sheet 61 to stimulate the cells in the culture solution to produce exosomes.

[0042] The reactor 5 is formed by combining a storage chamber 51, a filtration chamber 52, and a reaction chamber 53. After use, it is convenient to disassemble and clean. When installing the reactor 5, first insert the fixed plug 111 into the socket 114, and then the movable plug 112 is inserted into the socket 114 on the other side by the elastic force of the spring 113, thereby fixing the reactor 5 on the bracket 4. By installing the reactor 5 in the incubator 1, the temperature during culture is controlled, so that the cells are at a suitable growth temperature. After the cell culture cycle ends, the reactor 5 is inverted by rotating 180° through the driver 115, so that the culture solution flows down through the filtration layer 54 in the reaction chamber 53 for filtration and separation and then enters the storage chamber 51, thereby separating exosomes from other impurities such as organelles, making the whole culture process in a constant temperature state and without manual operation.

[0043] In the description of this specification, the description with reference to terms such as "one embodiment", "example", "specific example", etc. means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in a suitable manner in any one or more embodiments or examples.

[0044] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above embodiments, and what is described in the above embodiments and the specification only illustrates the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention will have various changes and improvements, and these changes and improvements all fall within the scope of the present invention claimed.

Claims

1. A culture device for promoting the secretion of exosomes from cells, comprising a constant temperature box (1), characterized in that: The interior of the incubator (1) is slidably connected to a culture platform (2), the culture platform (2) is fixedly connected to a vertical plate (3), the surface of the vertical plate (3) is rotatably connected to a U-shaped bracket (4) via an axis, the interior of the bracket (4) is detachably connected to a reactor (5) via a connecting component (11), the reactor (5) is mounted with a culture component (6), the culture component (6) comprises a plurality of conductive sheets (61) and a threaded connection end (63), the conductive sheet (61) is fixedly mounted at the bottom of the reactor (5), and the threaded connection end (63) is fixedly mounted at the bottom of the reactor (5). The threaded connection end (63) has an internal thread connected to a rotary joint (64), and the rotary joint (64) is fixedly connected to a three-way hose (65). A gas suction pump (7) and a gas output pump (8) are fixedly connected to the top of the culture platform (2). The two ends of the gas suction pump (7) and the gas output pump (8) are respectively fixedly connected to the two ends of the three-way hose (65). The two ends of the three-way hose (65) are respectively fixedly connected to an electromagnetic valve (9). An electric stimulation device (10) is fixedly connected to the culture platform (2), and the electric stimulation device (10) is electrically connected to the conductive sheet (61).

2. The culture device for promoting cell exosome secretion according to claim 1, characterized in that: The culture component (6) further comprises a current sensing device (62), wherein the current sensing device (62) is fixedly mounted on the bottom of the reactor (5), and the current sensing device (62) is electrically connected to the electrical stimulation device (10).

3. The culture device for promoting cell exosome secretion according to claim 1, characterized in that: A detection interface is installed on the surface of the rotary joint (64), and an air pressure sensor (66) is fixedly connected to the detection interface.

4. The culture device for promoting cell exosome secretion according to claim 1, characterized in that: The reactor (5) comprises a storage chamber (51), a filter chamber (52) and a reaction chamber (53) from top to bottom, and the storage chamber (51), the filter chamber (52) and the reaction chamber (53) are threadedly connected in sequence.

5. The culture device for promoting cell exosome secretion according to claim 4, characterized in that: A filter layer (54) is fixedly connected inside the filter chamber (52).

6. The culture device for promoting cell exosome secretion according to claim 5, characterized in that: The filter layer (54) is a hollow fiber filter membrane.

7. The culture device for promoting cell exosome secretion according to claim 1, characterized in that: The connection assembly (11) comprises a fixed plug (111) and a movable plug (112); the fixed plug (111) is fixedly mounted on one end of the bracket (4); and the movable plug (112) penetrates and is slidably mounted on the other end of the bracket (4).

8. The culture device for promoting cell exosome secretion according to claim 7, characterized in that: A spring (113) is sleeved on the surface of the movable plug (112).

9. The culture device for promoting cell exosome secretion according to claim 1, characterized in that: The connection assembly (11) further comprises two sockets (114), wherein the sockets (114) are fixedly mounted on both sides of the reactor (5).

10. The culture device for promoting cell exosome secretion according to claim 1, characterized in that: The connection assembly (11) further comprises a driver (115), wherein the driver (115) is fixedly mounted on the side wall of the vertical plate (3), and a driving end of the driver (115) is fixedly connected to the shaft of the bracket (4).

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