Exosome culture equipment
Through the design of the exogenous body culture equipment, the cell culture stent is used for 3D culture and simplifying the training solution transfer steps to solve the tedious problem of exosoma culture and improves the efficiency of exogenous output.
Patent Information
- Application Number
- CN202420522464.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-03-15
- Publication Date
- 2025-01-28
- Estimated Expiration
- 2034-03-15
AI Technical Summary
In the prior art, the exosome culture process is complicated, and frequent steps such as culture medium, transfer medium and separation are required, resulting in a low yield of exosomes.
A kind of exosocoman culture equipment is designed, which includes cell culture unit and exosocoman collection unit. It uses cell culture stent for 3D culture, and directly flows the cultivation solution through the valve to the exosocoman collection unit to simplify the process.
It improves the efficiency of exogenous output, reduces the frequency of steps in the cultivation process, and enhances the number of exosoma generated by a single culture.
Smart Images
Figure CN222411615U_ABST
Abstract
Description
Technical Field
[0001] The utility model relates to the field of biotechnology, and in particular to an exosome culture device. Background Art
[0002] As a cell product, exosomes are considered to be important carriers of cell-to-cell interaction. They are nanoscale vesicles secreted outside the cell by exocytosis. After the target cell absorbs the exosomes, its own function will be regulated. Stem cell-derived exosomes achieve cell-to-cell communication by delivering proteins, lipids and microRNAs (miRNAs). Stem cell-derived exosomes have been shown to have good therapeutic effects in wound healing, joint injury, fracture healing and heart injury, and have become an important raw material in the field of regenerative medicine and cell therapy. The culture of exosomes includes two steps: cell culture and exosome collection. In some technologies, cells are usually cultured on culture plates. After the cell culture is completed, the culture medium is taken out and then transferred to a separation device to collect the exosomes. However, since the number of cells cultured on the culture plate at a time is small, during the exosome culture process, it is necessary to frequently complete the steps of adding culture medium to the culture plate, transferring culture medium, and separating culture medium, which makes the culture of exosomes more cumbersome and makes the exosome yield low. Utility Model Content
[0003] The utility model aims to solve at least one of the technical problems existing in the prior art. To this end, the utility model proposes an exosome culture device that can improve the exosome output efficiency.
[0004] According to the exosome culture device, cell culture unit, exosome collection unit and valve of the embodiment of the utility model.
[0005] The cell culture unit includes a liquid preparation device, a culture plate and a cell culture support, the liquid preparation device has a liquid preparation cavity, the cell culture support has pores, the pores are used to accommodate cells, the culture plate has a culture hole and a first liquid discharge port connected to the culture hole, the culture hole is connected to the liquid preparation cavity, the cell culture support is arranged in the culture hole, the exosome collection unit includes a separation device and an exosome storage device, the separation device is connected to the first liquid discharge port, the exosome storage device includes a storage cavity, the storage cavity is connected to the separation device, and is used to store separated exosomes, and the valve is used to switch the flow channel between the cell culture unit and the exosome collection unit.
[0006] The exosome culture device according to the embodiment of the utility model has at least the following beneficial effects:
[0007] The cell culture unit of this embodiment is provided with a cell culture scaffold to realize 3D cell culture, thereby increasing the number of cells cultured at a single time, and after the cell culture is completed, the valve can be opened to allow the culture fluid to flow directly to the exosome collection unit to collect the exosomes, thereby improving the exosome production efficiency.
[0008] According to some embodiments of the utility model, the culture device also includes a culture chamber and a vortex generating device, the culture chamber is connected to the culture plate, the culture chamber defines a culture cavity, the culture cavity is connected to the liquid preparation cavity, the culture hole has an upward first liquid inlet, the first liquid inlet is connected to the culture cavity, and the vortex generating device is used to form a vortex in the culture fluid in the culture cavity.
[0009] According to some embodiments of the present invention, the culture bin has a containing groove and a notch connected to the containing groove, the notch is located at the bottom of the culture bin, and the culture plate is connected to the culture bin and covers the notch to define the culture cavity.
[0010] According to some embodiments of the utility model, the culture plate includes a plurality of culture wells, and the exosome culture equipment also includes a liquid collection device, the liquid collection device includes a liquid collection cavity, and a liquid collection port and a second liquid discharge port connected to the liquid collection cavity, the liquid collection port is located below each of the first liquid discharge ports, and is used to receive the culture fluid discharged through each of the first liquid discharge ports, and the second liquid discharge port is connected to the separation device.
[0011] According to some embodiments of the present invention, the cell culture unit further includes a filter assembly, the filter assembly includes a filter membrane, and the filter membrane is arranged at each of the first liquid discharge ports.
[0012] According to some embodiments of the utility model, the side wall of the culture bin has a protruding first abutment portion, the side of the liquid collecting device has a protruding second abutment portion, and the culture bin, the culture plate, the filter assembly and the liquid collecting device are sequentially distributed downward;
[0013] The exosome culture device also includes a connecting component, which includes a first connecting ring, a second connecting ring and a locking piece. The first connecting ring is sleeved on the side of the culture chamber and abuts against the side of the first abutting portion facing away from the liquid collection device. The second connecting ring is sleeved on the side of the liquid collection device and abuts against the side of the second abutting portion facing away from the culture chamber. The locking piece is connected to the first connecting ring and the second connecting ring so that the culture chamber, the culture plate, the filter assembly and the liquid collection device are clamped between the first connecting ring and the second connecting ring.
[0014] According to some embodiments of the utility model, the exosome culture equipment also includes a conveying device, which includes a peristaltic pump and a conveying hose, one end of the conveying hose is connected to the liquid preparation chamber, and the other end is connected to the culture hole, and the peristaltic pump is connected to the outer wall of the conveying hose.
[0015] According to some embodiments of the present invention, the separation device is a tangential flow filtration device, which includes a delivery pump, an ultrafiltration element and a liquid storage tank. The delivery pump, the ultrafiltration element and the liquid storage tank are connected in sequence to form a circulation pipeline, and the exosome storage device is connected to the ultrafiltration element.
[0016] According to some embodiments of the present invention, the exosome culture device also includes a temperature control component, which includes a heating element, a temperature sensor and a temperature controller. The heating element and the temperature sensor are both connected to the culture chamber and are communicatively connected to the temperature controller to adjust the temperature of the culture chamber.
[0017] According to some embodiments of the present invention, the heating element is configured as an annular structure, and the heating element is sleeved on the outer wall of the culture chamber.
[0018] Additional aspects and advantages of the present invention will be given in part in the following description, and in part will become apparent from the following description, or will be learned through the practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS
[0019] The present invention is further described below with reference to the accompanying drawings and embodiments, wherein:
[0020] Figure 1 This is a schematic diagram of the structure of an exosome culture device according to an embodiment of the utility model;
[0021] Figure 2 for Figure 1 A cross-sectional view of the culture device, the liquid collection device and the connection assembly;
[0022] Figure 3 A cross-sectional view of a culture plate, a filter assembly, and a cell culture support;
[0023] Figure 4 for Figure 2 Magnified view of area A in the middle;
[0024] Figure 5 This is a schematic diagram of the structure of another exosome culture device according to an embodiment of the present utility model.
[0025] Reference numerals:
[0026] Cell culture unit 10, exosome collection unit 20, valve 30;
[0027] Liquid dispensing device 100;
[0028] The culture device 200, the culture plate 210, the culture hole 211, the first liquid discharge port 212, the first liquid inlet 213, the culture chamber 220, the receiving groove 221, the first abutting portion 222, the culture chamber 230, the filter assembly 240, the filter membrane 241, the clamping member 242, the through hole 2421, and the heating member 250;
[0029] Cell culture support 300, separation device 400, delivery pump 410, ultrafiltration element 420, liquid storage tank 430;
[0030] Exosome storage device 500;
[0031] Agitator 600, stirring member 610;
[0032] Liquid collecting device 700, liquid collecting chamber 710, liquid collecting port 720, second liquid draining port 730, second abutting portion 740;
[0033] Connecting assembly 800, first connecting ring 810, second connecting ring 820, locking member 830;
[0034] Delivery device 900 , peristaltic pump 910 , delivery hose 920 . DETAILED DESCRIPTION
[0035] The embodiments of the present invention are described in detail below, and examples of the embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and cannot be understood as limiting the present invention.
[0036] In the description of the present invention, it should be understood that descriptions involving orientation, such as up, down, front, back, left, right, etc., indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention 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, and therefore cannot be understood as a limitation on the present invention.
[0037] In the description of the present utility model, "several" means more than one, "many" means more than two, "greater than", "less than", "exceed" etc. are understood to exclude the number itself, and "above", "below", "within" etc. are understood to include the number itself. If there is a description of "first" or "second", it is only used to distinguish the technical features, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.
[0038] In the description of the present invention, unless otherwise clearly defined, terms such as setting, installing, connecting, etc. should be understood in a broad sense, and technicians in the relevant technical field can reasonably determine the specific meanings of the above terms in the present invention based on the specific content of the technical solution.
[0039] In the description of the utility model, the description with reference to the terms "one embodiment", "some embodiments", "illustrative embodiments", "examples", "specific examples", or "some examples" means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the utility model. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.
[0040] As a cell product, exosomes are considered to be important carriers of cell-to-cell interaction. They are nanoscale vesicles secreted outside the cell by exocytosis. After the target cell absorbs the exosomes, its own function will be regulated. Stem cell-derived exosomes achieve cell-to-cell communication by delivering proteins, lipids and microRNAs (miRNAs). Stem cell-derived exosomes have been shown to have good therapeutic effects in wound healing, joint injury, fracture healing and heart injury, and have become an important raw material in the field of regenerative medicine and cell therapy. The culture of exosomes includes two steps: cell culture and exosome collection. In some technologies, cells are usually cultured on culture plates. After the cell culture is completed, the culture medium is taken out and then transferred to a separation device to collect the exosomes. However, since the number of cells cultured on the culture plate at a time is small, during the exosome culture process, it is necessary to frequently complete the steps of adding culture medium to the culture plate, transferring culture medium, and separating culture medium, which makes the culture of exosomes more cumbersome and makes the exosome yield low.
[0041] Based on the above problems, the utility model proposes an exosome culture device, referring to Figures 1 to 3 , Figure 1 This is a schematic diagram of the structure of an exosome culture device according to an embodiment of the utility model. Figure 2 for Figure 1 A cross-sectional view of the culture device, the liquid collection device and the connection components, Figure 3 It is a cross-sectional view of a culture plate, a filter assembly and a cell culture support. The exosome culture device of this embodiment includes a cell culture unit 10 , an exosome collection unit 20 and a valve 30 .
[0042] The cell culture unit 10 includes a liquid preparation device 100, a culture plate 210, and a cell culture support 300. The liquid preparation device 100 has a liquid preparation chamber, which is used to prepare the culture solution and temporarily store the culture solution. The culture plate 210 has a culture hole 211 and a first liquid discharge port 212 connected to the culture hole 211. The culture hole 211 is connected to the liquid preparation chamber. During the exosome culture process, the culture solution flows into the culture hole 211 to provide the cells with the required nutrients. Specifically, for example, there is also a conveying device 900 between the liquid preparation chamber and the culture hole 211. When liquid supply is required, the conveying device 900 is started to pump the culture solution into the culture hole 211.
[0043] A cell culture support 300 (such as Figure 3 As shown, the cell culture scaffold 300 has pores, and the pores are used to accommodate cells. Taking stem cells extracted from umbilical cord or adipose tissue as an example, the seed cells are amplified to P3-P4 generations through cell microcarriers and then retained for standby use. A mixed solution of gelatin and sodium alginate hydrogel is prepared in advance with physiological saline, wherein the concentration range of gelatin is 5%-15% (mass fraction), and the concentration range of sodium alginate is 1%-4% (mass fraction). The hydrogel mixed solution is mixed with stem cells to form biological ink, and finally the biological ink is printed into a calcium chloride solution through a 3D printing device, and the calcium chloride solution is soaked for cross-linking and curing to form a cell culture scaffold 300 containing cells in a grid or the like. That is, when manufacturing the cell culture scaffold 300, the cells are wrapped by the biomaterial and embedded in the cell culture scaffold 300. Alternatively, a cell culture scaffold 300 is first made separately, and then the cells are planted on the surface of the scaffold so that the cells naturally migrate to the pores of the scaffold. When culturing cells, the cell culture scaffold 300 containing cells or attached with cells can be directly placed in the culture hole 211.
[0044] The exosome collection unit 20 includes a separation device 400 and an exosome storage device 500. The separation device 400 is connected to the first liquid discharge port 212. The exosome storage device 500 includes a storage cavity, which is connected to the separation device 400. The separation device 400 is, for example, a centrifuge or a filter for storing separated exosomes. The valve 30 is used to switch the flow channel between the cell culture unit 10 and the exosome collection unit 20. For example, the valve 30 is set at the first liquid discharge port 212, but it is not limited thereto. It can also be set in a delivery pipe or other structure (such as the liquid collection device 700 described below). During the cell culture process, the valve 30 is first closed. After the cell culture is completed, the valve 30 is opened again to allow the culture fluid in the culture chamber to flow directly into the separation device 400 through the first liquid discharge port 212 without first taking out the culture fluid, thereby simplifying the steps of transferring the culture fluid and improving the efficiency of transferring the culture fluid. In addition, the cell culture unit 10 uses a cell culture scaffold 300 to perform 3D culture on cells, thereby increasing the number of cells cultured per unit volume, thereby increasing the number of exosomes secreted by the cells. Therefore, compared with the conventional technology, the exosome culture device of this embodiment not only improves the transfer efficiency of the culture fluid, but also increases the number of exosomes produced in a single culture, thereby eliminating the need to frequently perform steps such as adding culture fluid, transferring culture fluid, and separating culture fluid during the culture process, thereby improving the exosome production efficiency.
[0045] In addition, it should be noted that in this embodiment, since the cells are contained in the pores of the cell culture support 300, only a small number of cells will be free in the culture medium during the cell culture process. Therefore, when the culture medium is transferred to the exosome collection unit 20, only a small number of cells will enter the separation device 400 with the culture medium, while most of the cells will remain in the cell culture support 300, thereby allowing most of the cells to be recycled, thereby saving costs.
[0046] Reference Figure 2 In some embodiments, the culture device 200 further includes a culture chamber 220 and a vortex generating device. The culture chamber 220 defines a culture chamber 230, the culture chamber 230 is connected to the liquid preparation chamber, the culture hole 211 has a first liquid inlet 213, the first liquid inlet 213 is connected to the culture chamber 230, and the vortex generating device is connected to the culture chamber 220, and is used to form a vortex in the culture liquid in the culture chamber 230, so that the culture liquid generates fluid shear force. For example, the vortex generating device is a vortex generator, a vortex generator, or a stirrer 600, etc., Figure 2 As shown in the figure, the vortex generating device is a stirrer 600, which includes a stirring member 610 and a driving mechanism. The stirring member 610 partially extends into the culture chamber 230. The driving mechanism is connected to the stirring member 610 and is used to drive the stirring member 610 to rotate, thereby driving the culture fluid to rotate and generate fluid shear force, thereby causing the cells to secrete more exosomes.
[0047] In addition, it should be noted that, compared with direct culture using a culture plate in conventional techniques, the cells in this embodiment are located in a cell culture support 300 and are not easily damaged. Therefore, in this embodiment, the vortex generator can generate a faster vortex, thereby increasing the amount of exosome secretion.
[0048] Based on the above embodiment, for example, the vortex generating device is set as a stirrer 600, and the rotation speed of the stirring member 610 is 60r / min to 150r / min. Specifically, it can be understood that when the rotation speed of the stirring member 610 is too large, a large fluid shear force will be formed, resulting in cell damage, and when the rotation speed of the stirring member 610 is too small, the fluid shear force formed is small, resulting in a decrease in the output of exosomes. Based on this, in this embodiment, the rotation speed of the stirring member 610 is set within a reasonable range, which ensures that the cells are not damaged, so that the cells can secrete more exosomes.
[0049] Based on the above embodiment, the culture chamber 230 may be formed inside the culture chamber 220, and the culture plate 210 may be disposed in the culture chamber 230 so that the culture hole 211 is connected to the culture chamber 230. Alternatively, the culture chamber 220 may have a receiving groove 221 and a notch connected to the receiving groove 221, the notch being located at the bottom of the culture chamber 220, and the culture plate 210 may be connected to the culture chamber 220 and cover the notch to define the culture chamber 230 (e.g., Figure 2 That is, in this embodiment, the culture plate 210 is used as the bottom wall of the culture chamber 230, so that the structure of the exosome culture plate 210 of this embodiment is simpler, saving equipment manufacturing costs.
[0050] Reference Figure 2 In some embodiments, the culture plate 210 includes a plurality of culture wells 211, and the exosome culture device further includes a liquid collection device 700, which includes a liquid collection chamber 710, and a liquid collection port 720 and a second liquid discharge port 730 connected to the liquid collection chamber 710. The liquid collection port 720 is located below each first liquid discharge port 212 and is used to receive the culture fluid discharged through each first liquid discharge port 212. The second liquid discharge port 730 is connected to the separation device 400. Figure 2 As shown in the example, the liquid collecting device 700 is funnel-shaped, and the liquid collecting device 700 is buckled at the bottom of the culture device 200, and each first liquid discharge port 212 is located within the range of the liquid collecting port 720. Therefore, when the culture liquid is discharged from the culture hole 211, the culture liquid discharged from each first liquid discharge port 212 can flow into the liquid collecting cavity 710, and then flow to the separation device 400 through the second liquid discharge port 730. That is, there is no need for one first liquid discharge port 212 to correspond to one delivery pipeline, so that the structure of the exosome culture device of this embodiment is simpler, saving equipment cost.
[0051] Reference Figure 2 and Figure 3 In some embodiments, the cell culture unit 10 further includes a filter assembly 240, the filter assembly 240 includes a filter membrane 241, and the filter membrane 241 is disposed at each first liquid discharge port 212. Specifically, for example, the filter assembly 240 includes a clamp 242, the clamp 242 has a through hole 2421, the filter membrane 241 covers the through hole 2421, the clamp 242 is connected to the bottom of the culture plate 210, so that the filter membrane 241 is clamped between the culture plate 210 and the clamp 242. Further, in some embodiments, the upper surface of the clamp 242 has a boss and a positioning column, the lower surface of the culture plate 210 has a mounting groove and a positioning hole, and the filter membrane 241 is disposed at In the mounting groove, the clamp 242 is mounted on the bottom of the culture plate 210, and the boss is extended into the mounting groove to clamp the filter membrane 241, and the positional accuracy between the clamp 242 and the culture plate 210 is improved by the cooperation of the positioning column and the positioning hole, so that the through hole 2421 on the clamp 242 corresponds to the first liquid discharge port 212 of the culture plate 210, so that the culture fluid is discharged from the first liquid discharge port 212, filtered through the filter membrane 241 and flows from the through hole 2421 to the liquid collection chamber 710. Specifically, different adapted filter membranes 241 are selected according to the cultured cells. When the culture fluid passes through the filter membrane 241, the filter membrane 241 can separate most of the cells in the culture fluid and the particles formed by the degradation of the cell culture support 300. In other words, the filter membrane 241 in this embodiment is used to perform primary separation of the culture fluid, and then the separation device 400 is used to perform secondary separation of the culture fluid, thereby improving the separation efficiency of the separation device 400 for the culture fluid, and then improving the exosome culture efficiency.
[0052] In addition, it should be noted that it is understandable that the filter membrane 241 is arranged at the first drain port 212, so that the cell culture bracket 300 can be directly arranged on the filter membrane 241, that is, the cell culture bracket 300 is supported by the filter membrane 241 without the need to set up other additional supporting structures, thereby making the structure simpler and reducing the equipment cost.
[0053] Reference Figure 2 and Figure 4 , Figure 4 for Figure 2An enlarged view of the area A in the middle. In some embodiments, the side wall of the culture chamber 220 has a protruding first abutment 222, and the side of the liquid collection device 700 has a protruding second abutment 740. The culture chamber 220, the culture plate 210, the filter assembly 240, and the liquid collection device 700 are sequentially distributed downward. The exosome culture device also includes a connecting assembly 800, which includes a first connecting ring 810, a second connecting ring 820, and a locking member 830. The first connecting ring 810 is sleeved on the side of the culture chamber 220 and abuts against the side of the first abutment 222 that is away from the liquid collection device 700. The second connecting ring 820 is sleeved on the side of the liquid collection device 700 and abuts against the side of the second abutment 740 that is away from the culture chamber 220. The locking member 830 is connected to the first connecting ring 810 and the second connecting ring 820 so that The culture chamber 220, the culture plate 210, the filter assembly 240 and the liquid collecting device 700 are clamped between the first connecting ring 810 and the second connecting ring 820. That is, in this embodiment, the installation of the culture chamber 220, the culture plate 210, the filter assembly 240 and the liquid collecting device 700 can be completed through the connecting assembly 800, so that the structure of the exosome culture device of this embodiment is simpler, thereby reducing the equipment cost, and the construction process is simpler and more convenient, making the culture use and maintenance of this embodiment more convenient.
[0054] Furthermore, in order to improve the sealing of the device, some embodiments also include sealing members, and sealing members are provided between the culture chamber 220 and the culture plate 210 , between the culture plate 210 and the clamp 242 , and between the clamp 242 and the liquid collection device 700 .
[0055] Reference Figure 1 In some embodiments, the cell culture unit 10 further includes a conveying device 900, the conveying device 900 includes a peristaltic pump 910 and a conveying hose 920, one end of the conveying hose 920 is connected to the liquid dispensing chamber, and the other end is connected to the culture hole 211, and the peristaltic pump 910 is connected to the outer wall of the conveying hose 920. Specifically, for example, the culture chamber 220 is plugged with a hard connecting tube, one end of the conveying hose 920 is connected to the liquid dispensing device 100, and the other end is connected to the hard connecting tube, and the peristaltic pump 910 conveys the culture solution to the culture hole 211 by squeezing the conveying hose 920. Therefore, during the process of conveying the culture solution, the culture solution only contacts the conveying hose 920, thereby making it easier and more convenient to clean and disinfect the conveying device, thereby making the exosome culture device of this embodiment more convenient to use. In addition, compared with an ordinary conveying pump, the fluid shear force generated by the peristaltic pump 910 during the process of conveying the culture solution is lower, thereby preventing the nutrients in the culture solution from being destroyed.
[0056] Reference Figure 5 , Figure 5This is a schematic diagram of the structure of another exosome culture device according to an embodiment of the utility model. In some embodiments, the separation device 400 is configured as a tangential flow filtration device, which includes a delivery pump 410, an ultrafiltration element 420, and a liquid storage tank 430. The delivery pump 410 may be, for example, a peristaltic pump. The delivery pump 410, the ultrafiltration element 420, and the liquid storage tank 430 are connected through a pipeline to form a circulating filtration pipeline. The exosome storage device 500 is connected to the ultrafiltration element 420, and the pipeline between the delivery pump 410 and the storage tank is connected to the cell culture unit 10 through a tee. During the exosome culture process, when the cell culture is completed, the valve 30 is opened to allow the culture solution to flow into the liquid storage tank 430, and the delivery pump 410 is started to circulate and filter the culture solution, thereby increasing the filtration speed.
[0057] In some embodiments, the exosome culture device further includes a temperature control component, which includes a heating element 250, a temperature sensor and a temperature controller. The heating element 250 is connected to the culture chamber 220 and is connected to the temperature controller to adjust the temperature of the culture hole 211. Specifically, the temperature sensor is used to detect the temperature of the culture solution in the culture chamber 230, that is, the temperature of the cell culture environment, and transmit the temperature information to the temperature controller, which is, for example, a PID temperature controller. The temperature controller controls the heating element 250 to cool down or heat up to adjust the temperature of the culture solution, so that the temperature of the culture solution reaches a suitable temperature for the cultured cells, thereby increasing the number of exosomes secreted by the cells.
[0058] Based on the above embodiment, the heating element 250 is configured as a ring structure, and the heating element 250 is sleeved on the outer wall of the culture chamber 220, so that the temperature of the culture solution is more uniform, thereby increasing the number of exosomes secreted by the cells.
[0059] In addition, in some embodiments, the culture device 200 further includes a CO2 monitoring component, which includes a CO2 sensor and a CO2 controller. The CO2 sensor is disposed in the culture chamber 230 and is communicatively connected to the CO2 controller. Specifically, the 5% CO2 mixed gas is connected to the CO2 interface of the culture chamber 220 through a stop valve, a gas solenoid valve, and an air filter, and flows out from the CO2 outlet into the culture chamber 230. The CO2 sensor monitors the CO2 concentration in the culture chamber 230 and transmits the detection information to the CO2 controller. The CO2 controller compares the detection information with the set value range of 5% ± 0.2%, feeds back the difference to the CO2 controller, and controls the increase or decrease of the 5% CO2 mixed gas through the gas solenoid valve.
[0060] The embodiments of the utility model are described in detail above in conjunction with the accompanying drawings, but the utility model is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the utility model. In addition, the embodiments of the utility model and the features in the embodiments can be combined with each other without conflict.
Claims
1. An exosome culture device, characterized in that: include: A cell culture unit, comprising a liquid preparation device, a culture device and a cell culture support, wherein the liquid preparation device has a liquid preparation cavity, the culture device comprises a culture plate, the culture plate has a culture hole and a first liquid discharge port connected to the culture hole, the culture hole is connected to the liquid preparation cavity, the cell culture support has a pore for accommodating cells, and the cell culture support is arranged in the culture hole; An exosome collection unit, comprising a separation device and an exosome storage device, wherein the separation device is connected to the first liquid discharge port, and the exosome storage device has a storage cavity, which is connected to the separation device and is used to store the separated exosomes; A valve is used to switch the flow channel between the cell culture unit and the exosome collection unit.
2. The exosome culture device according to claim 1, characterized in that: The culture device also includes a culture chamber and a vortex generating device, wherein the culture chamber is connected to the culture plate, the culture chamber defines a culture cavity, the culture cavity is connected to the liquid preparation cavity, the culture hole has an upward first liquid inlet, the first liquid inlet is connected to the culture cavity, and the vortex generating device is used to form a vortex in the culture liquid in the culture cavity.
3. The exosome culture device according to claim 2, characterized in that: The culture bin has a containing groove and a notch communicating with the containing groove, wherein the notch is located at the bottom of the culture bin, and the culture plate is connected to the culture bin and covers the notch to define and form the culture cavity.
4. The exosome culture device according to claim 3, characterized in that: The culture plate includes a plurality of culture wells, and the exosome culture equipment also includes a liquid collection device, which includes a liquid collection cavity, and a liquid collection port and a second liquid discharge port connected to the liquid collection cavity, the liquid collection port is located below each of the first liquid discharge ports, and is used to receive the culture fluid discharged through each of the first liquid discharge ports, and the second liquid discharge port is connected to the separation device.
5. The exosome culture device according to claim 4, characterized in that: The culture device further comprises a filter assembly, wherein the filter assembly comprises a filter membrane, and the filter membrane is arranged at each of the first liquid discharge ports.
6. The exosome culture device according to claim 5, characterized in that: The side wall of the culture chamber has a protruding first abutment portion, the side of the liquid collecting device has a protruding second abutment portion, and the culture chamber, the culture plate, the filter assembly and the liquid collecting device are sequentially distributed downward; The exosome culture device also includes a connecting component, which includes a first connecting ring, a second connecting ring and a locking piece. The first connecting ring is sleeved on the side of the culture chamber and abuts against the side of the first abutting portion facing away from the liquid collection device. The second connecting ring is sleeved on the side of the liquid collection device and abuts against the side of the second abutting portion facing away from the culture chamber. The locking piece is connected to the first connecting ring and the second connecting ring so that the culture chamber, the culture plate, the filter assembly and the liquid collection device are clamped between the first connecting ring and the second connecting ring.
7. The exosome culture device according to claim 1, characterized in that: The cell culture unit also includes a conveying device, which includes a peristaltic pump and a conveying hose. One end of the conveying hose is connected to the liquid preparation cavity, and the other end is connected to the culture hole. The peristaltic pump is connected to the outer wall of the conveying hose.
8. The exosome culture device according to claim 1, characterized in that: The separation device is a tangential flow filtration device, which includes a delivery pump, an ultrafiltration element and a liquid storage tank. The delivery pump, the ultrafiltration element and the liquid storage tank are connected in sequence to form a circulation pipeline, and the exosome storage device is connected to the ultrafiltration element.
9. The exosome culture device according to claim 2, characterized in that: The culture device also includes a temperature control component, which includes a heating element, a temperature sensor and a temperature controller. The heating element and the temperature sensor are both connected to the culture chamber and are both communicatively connected to the temperature controller to adjust the temperature of the culture chamber.
10. The exosome culture device according to claim 9, characterized in that: The heating element is configured as a ring structure, and the heating element is sleeved on the outer wall of the culture chamber.