Brown adipocyte in-vitro culture device

Abstract

The invention provides a brown adipocyte in-vitro culture device, and relates to the field of adipocyte in-vitro culture, the brown adipocyte in-vitro culture device comprises a frame body, and an amplification assembly used for cell amplification and a separation assembly used for cell acquisition are spaced up and down in the frame body; the amplification assembly comprises a mounting plate arranged in an inner cavity of the frame body, rotating plates are arranged on the outer side of the mounting plate in a rectangular array mode, test tubes sleeve the middles of the rotating plates, a culture dish for cell culture sleeves the middle of the mounting plate, a cover plate is arranged at the top of the culture dish, and the cover plate is connected with the rotating plates. And the top of the cover plate is in pipe connection with a second sleeve. The frame body is sterilized through the arranged sterilization pipe and the sterilization nozzle installed at one end of the sterilization pipe, precursor cells are extracted through the separation assembly, cell amplification and induction are conducted in cooperation with the amplification assembly, automatic induction cultivation is achieved, sterilization operation is conducted, and bacterial infection is avoided.

Description

Technical Field

[0001] The present invention relates to the field of in vitro culture of fat cells, and in particular to an in vitro culture device for brown fat cells. Background Art

[0002] Obesity has become a major public health issue worldwide, increasing the risk of cardiovascular disease, diabetes, metabolic syndrome and other diseases; traditional weight loss methods, such as diet control, exercise and drug therapy, often fail to achieve long-term results; brown adipose tissue (BAT) is considered a new way to treat obesity because it can consume excess energy through non-shivering heat production. Currently, when culturing brown fat cells, the existing cell culture method is a single culture dish with a cover plate using ceiling technology. This culture method is not convenient for classification and requires excessive human intervention, which makes bacterial infection easy to occur during the culture process.

[0003] For example: The utility model patent (application number: CN202021400267.7) discloses a "fat cell culture device", and its specification discloses: the traditional culture dish consists of a single flat disc-shaped bottom and a cover, which is difficult to classify and culture, and has poor sealing. After the culture solution is injected, it is difficult to vibrate or shake it to make it evenly distributed. It is inconvenient to operate and has few functions. The above patent can prove the defects of the prior art.

[0004] Therefore, we made improvements on this and proposed an in vitro culture device for brown adipocytes. Summary of the invention

[0005] The purpose of the present invention is to address the current problem that when culturing brown fat cells, the existing cell culture method is to use a single culture dish with a cover plate using ceiling technology for culture. This culture method is not convenient for classified culture and requires excessive human intervention, which leads to the problem of bacterial infection during the culture process.

[0006] In order to achieve the above-mentioned purpose of the invention, the present invention provides the following in vitro culture device for brown adipocytes to improve the above-mentioned problems.

[0007] The specific application is as follows: A brown adipocyte in vitro culture device comprises a frame, wherein an amplification component for cell amplification and a separation component for cell acquisition are arranged in the upper and lower parts of the frame; The amplification component includes a mounting plate arranged in the inner cavity of the frame, and a rotating plate is provided in a rectangular array outside the mounting plate, and a test tube is sleeved in the middle of the rotating plate, and a culture dish for cell culture is sleeved in the middle of the mounting plate, a cover plate is provided on the top of the culture dish, and a second sleeve is connected to the top of the cover plate.

[0008] As a preferred technical solution of the present application, a mounting frame is sleeved on the middle part of the outer side of the second sleeve, and the bottom of the mounting frame is fixedly connected to the top of the mounting plate, a mounting pad is fixedly installed on the bottom of the inner cavity of the second sleeve, and the mounting pad is annular, an extrusion spring is fixedly installed on the top of the mounting pad, and a slider is fixedly installed on the top of the extrusion spring, and the second sleeve is movably sleeved on the slider.

[0009] As a preferred technical solution of the present application, the inner cavity of the slider is provided with an inner groove, and the inner groove is in a right angle shape, the bottom of the inner groove passes through the bottom of the slider, and the other end of the inner groove passes through the side wall of the slider, and the top tube of the side wall of the second sleeve is connected to a feeding pipe, and the feeding pipe is connected to the inner groove.

[0010] As a preferred technical solution of the present application, a first connecting block is sleeved on the top of the slider, and a pull rope is provided in a rectangular array on the top of the first connecting block. A second connecting block is arranged above the first connecting block at intervals, and a fixed block is sleeved on the top of the second connecting block, and the top of the fixed block is fixedly connected to the top of the inner cavity of the frame.

[0011] As a preferred technical solution of the present application, the pull rope passes through the bottom of the second connecting block and extends out of its side wall, and the other end of the second connecting block is fixedly connected to the top of the rotating plate.

[0012] As a preferred technical solution of the present application, the separation component includes a pressure-stabilizing box arranged at the bottom of the culture dish, the top of the pressure-stabilizing box is fixedly connected to the mounting plate, the side walls of the pressure-stabilizing box are provided with grooves, and the pressure-stabilizing box is connected to the test tube through the grooves, a connecting box is provided at the bottom of the pressure-stabilizing box, and the connecting box is connected to the pressure-stabilizing box, the side wall array pipe of the connecting box is connected with a connecting pipe, and the outer side of the connecting pipe is provided with a valve body.

[0013] As a preferred technical solution of the present application, the bottom pipe of the connection box is connected to a transmission disc, the transmission disc is tubular, and the outer side of the bottom of the transmission disc is gear-shaped, the middle part of the transmission disc has a valve body, the bottom of the transmission disc is sleeved with a funnel, and the bottom pipe of the funnel is connected to an air pump.

[0014] As a preferred technical solution of the present application, a driving motor is spaced apart on one side of the air pump, and a main gear is fixedly mounted on the output end of the driving motor, and the main gear is meshed with the bottom of the transmission disc.

[0015] As a preferred technical solution of the present application, the terminal tube of the connecting tube is connected to a first sleeve, and an arc-shaped sleeve is fixedly installed at one end of the first sleeve, and a sterilization tube extending outward is provided at the other end of the first sleeve, and one end of the sterilization tube extending out of the first sleeve is made of rubber, and the sterilization tube is in contact with the outside of the test tube, and a sterilization nozzle is penetrated at one end of the sterilization tube.

[0016] As a preferred technical solution of the present application, one end of the arc sleeve is fixedly connected to the bottom of the pressure stabilizing box, the other end of the arc sleeve is sleeved with a transmission tube extending outward, and the transmission tube is fixedly connected to the bottom of the rotating plate, the rotation center of the transmission tube and the arc sleeve is located on the rotation axis of the rotating plate, the other end of the transmission tube is fixedly installed with a reset spring, and the transmission tube is connected to the inner wall of the arc sleeve through the reset spring, and the separation component and the amplification component adopt an in vitro culture method.

[0017] Compared with the prior art, the present invention has the following beneficial effects: In the scheme of this application: 1. In order to solve the problem of bacterial infection of the culture dish caused by manual intervention when the culture dish is combined with the cover plate for ceiling induction monoculture in the prior art, the present application sterilizes the frame by providing a sterilization tube and a sterilization nozzle installed at one end of the sterilization tube, extracts precursor cells by separation components, and amplifies and induces cells by cooperating with the amplification components, thereby realizing automated induction cultivation and performing sterilization operations to avoid bacterial infection; 2. In order to solve the problem that the culture method in the prior art is not convenient for classified culture, the present application adds partitions inside the culture dish to divide the culture dish, thereby realizing classified culture of cells by changing the culture dish and adapting different test tubes; 3. The slider is provided in combination with the feeding tube, which makes it easy to add culture fluid to the inside of the culture dish, and the second sleeve is sealed by the slider, so that different culture media can be added to the inside of the culture dish, solving the problem of bacterial infection that is easy to occur when adding culture media in the prior art. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 A schematic diagram of the structure of the in vitro culture device for brown adipocytes provided in this application; Figure 2 A structural cross-sectional view of the frame of the brown adipocyte in vitro culture device provided in this application; Figure 3 A schematic diagram of the internal structure of the frame of the in vitro culture device for brown adipocytes provided in this application; Figure 4 A schematic diagram of the connection structure of the mounting plate of the in vitro culture device for brown adipocytes provided in this application; Figure 5 A schematic cross-sectional view of the voltage stabilizing box of the brown adipocyte in vitro culture device provided in the present application; Figure 6 A schematic diagram of the connection structure of the connection box of the brown adipocyte in vitro culture device provided in this application; Figure 7 A schematic diagram of the connection structure of the first sleeve of the brown adipocyte in vitro culture device provided in this application; Figure 8 A cross-sectional view of the internal structure of the arc-shaped sleeve of the in vitro culture device for brown adipocytes provided in the present application; Fig. 9 A cross-sectional view of the connection structure of the first sleeve of the brown adipocyte in vitro culture device provided in the present application; Fig.10 A cross-sectional view of the connection structure of the connection box of the brown adipocyte in vitro culture device provided in this application; Fig.11 A schematic diagram of the structure of the expansion component of the in vitro culture device for brown adipocytes provided in this application; Fig.12 A schematic cross-sectional view of the second sleeve of the brown adipocyte in vitro culture device provided in the present application; Fig.13 Schematic diagram of the cross-sectional structure of the slider of the brown adipocyte in vitro culture device provided in this application Fig.14 A flow chart of the in vitro culture method of brown adipocytes provided in this application.

[0019] Indicated in the figure: 1. Frame; 2. Separation assembly; 201. Drive motor; 202. Main gear; 203. Connecting box; 204. Transmission plate; 205. Voltage stabilizer box; 206. Connecting pipe; 207. First sleeve; 208. Sterilization tube; 209. Sterilization nozzle; 210. Arc sleeve; 211. Transmission tube; 212. Return spring; 213. Air pump; 214. Funnel; 3. Amplification component; 301. Mounting plate; 302. Rotating plate; 303. Test tube; 304. Culture dish; 305. Cover plate; 306. Second sleeve; 307. Mounting pad; 308. Extrusion spring; 309. Slider; 310. Inner groove; 311. First connecting block; 312. Pull rope; 313. Second connecting block; 314. Fixed block; 315. Feeding tube; 316. Mounting bracket. DETAILED DESCRIPTION

[0020] In order to enable those skilled in the art to better understand the scheme of the present invention, the technical scheme in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0021] As described in the background art, when brown fat cells are cultured, the existing cell culture method is to use a single culture dish with a cover plate using ceiling technology. This culture method is not convenient for classified culture and requires excessive human intervention, which leads to the problem of bacterial infection during the culture process.

[0022] In order to solve this technical problem, the present invention provides a brown adipocyte in vitro culture device, which is applied to automated aseptic cell culture.

[0023] Specifically, please refer to Figure 1-Figure 14 The brown adipocyte in vitro culture device specifically comprises a frame 1, wherein an amplification component 3 for cell amplification and a separation component 2 for cell acquisition are arranged in the upper and lower parts of the frame 1; The amplification component 3 includes a mounting plate 301 arranged in the inner cavity of the frame 1, and a rotating plate 302 is provided in a rectangular array outside the mounting plate 301, the mounting plate 301 is sleeved with the rotating plate 302, and the rotating plate 302 is sleeved with the mounting plate 301 through a torsion spring, and a test tube 303 is sleeved in the middle of the rotating plate 302, a culture dish 304 for cell culture is sleeved in the middle of the mounting plate 301, a cover plate 305 is provided on the top of the culture dish 304, and a second sleeve 306 is connected to the top of the cover plate 305.

[0024] The brown adipocyte in vitro culture device provided by the present invention sterilizes the frame by means of a sterilization tube and a sterilization nozzle installed at one end of the sterilization tube, extracts precursor cells by means of a separation component, and then cooperates with an amplification component to amplify and induce the cells, thereby realizing automated induction cultivation and performing sterilization operations to avoid bacterial infection.

[0025] In order to enable those skilled in the art to better understand the solutions of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings.

[0026] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features and technical solutions in the embodiments may be combined with each other.

[0027] 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, further definition and explanation thereof is not required in subsequent drawings.

[0028] Example 1, please refer to Figure 3 , Figure 4 , Fig.11 , Fig.12 and Fig.13 , a brown fat cell in vitro culture device, wherein a mounting frame 316 is sleeved on the middle part of the outer side of the second sleeve 306, and the bottom of the mounting frame 316 is fixedly connected to the top of the mounting plate 301, a mounting pad 307 is fixedly installed on the bottom of the inner cavity of the second sleeve 306, and the mounting pad 307 is annular, a squeezing spring 308 is fixedly installed on the top of the mounting pad 307, and a slider 309 is fixedly installed on the top of the squeezing spring 308, the second sleeve 306 and the slider 309 are movably sleeved, the inner cavity of the slider 309 is provided with an inner groove 310, and the inner groove 310 is in a right angle shape, the bottom of the inner groove 310 passes through the bottom of the slider 309, and the other end of the inner groove 310 passes through the side wall of the slider 309, the top of the side wall of the second sleeve 306 is connected with a feeding pipe 315, and the feeding pipe 315 is connected with the inner groove 310; By providing an extrusion spring 308 in cooperation with the slider 309 that is slidably sleeved with the second sleeve 306, it is convenient to limit and reset the slider 309 when it moves, and the inner groove 310 opened in the inner cavity of the slider 309 is connected with the feeding tube 315, so that the culture medium can be conveniently added to the inner cavity of the culture dish 304 through the second sleeve 306 and the cover plate 305.

[0029] Please refer to Figure 3 , Figure 4 , Fig.11 , Fig.12 and Fig.13 , a brown adipocyte in vitro culture device, wherein a first connecting block 311 is sleeved on the top of a slider 309, and a drawstring 312 is arranged in a rectangular array on the top of the first connecting block 311, a second connecting block 313 is arranged above the first connecting block 311, and a fixing block 314 is sleeved on the top of the second connecting block 313, the top of the fixing block 314 is fixedly connected to the top of the inner cavity of the frame 1, the drawstring 312 passes through the bottom of the second connecting block 313 and extends out of its side wall, and the other end of the second connecting block 313 is fixedly connected to the top of the rotating plate 302; By rotating the rotating plate 302, the pull rope 312 is pulled, and under the action of the extrusion spring 308, the slider 309 is pulled by the pull rope 312 to slide inside the second sleeve 306. When the inner groove 310 opened in the inner cavity of the slider 309 cooperates with the feeding tube 315, the culture medium can be added conveniently.

[0030] Example 2 further optimizes the in vitro culture device for brown adipocytes provided in Example 1. Specifically, Figure 3 , Figure 4 , Figure 5 , Figure 6 , Figure 7 , Figure 8 , Fig. 9 ,and Fig.10 As shown, the separation component 2 includes a pressure stabilizing box 205 arranged at the bottom of the culture dish 304, the top of the pressure stabilizing box 205 is fixedly connected to the mounting plate 301, the side walls of the pressure stabilizing box 205 are provided with grooves, and the pressure stabilizing box 205 is sleeved with the test tube 303 through the grooves, the bottom of the pressure stabilizing box 205 is provided with a connecting box 203, and the connecting box 203 is connected with the pressure stabilizing box 205, the side wall array pipe of the connecting box 203 is connected with a connecting pipe 206, and the outer side of the connecting pipe 206 is provided with a valve body; The voltage stabilizing box 205 is used to support the culture dish 304 and the mounting plate 301 , and the voltage stabilizing box 205 is sleeved with the test tube 303 , so as to limit the position of the test tube 303 .

[0031] Furthermore, Figure 3 , Figure 4 , Figure 5 , Figure 6 , Figure 7 , Figure 8 , Fig. 9 ,and Fig.10 As shown, the bottom pipe of the connection box 203 is connected with a transmission disc 204, the transmission disc 204 is tubular, and the outer side of the bottom of the transmission disc 204 is gear-shaped, the middle of the transmission disc 204 is provided with a valve body, the bottom of the transmission disc 204 is sleeved with a funnel 214, and the bottom pipe of the funnel 214 is connected with an air pump 213, a driving motor 201 is arranged at intervals on one side of the air pump 213, and the output end of the driving motor 201 is fixedly installed with a main gear 202, and the main gear 202 is meshed with the bottom of the transmission disc 204; By means of the air pump 213 and the funnel 214 connected to the air pump 213, it is convenient to extract and discharge gas into the transmission disc 204, the connection box 203 and the pressure stabilizing box 205, thereby changing the internal air pressure, and stabilizing the pressure through the pressure stabilizing box 205 to ensure pressure synchronization. The driving motor 201 drives the transmission disc 204, the connection box 203, the pressure stabilizing box 205 and the mounting plate 301 to rotate through the main gear 202 installed at its output end, thereby realizing centrifugal operation.

[0032] Furthermore, Figure 3 , Figure 4 , Figure 5 , Figure 6 , Figure 7 , Figure 8 , Fig. 9 ,and Fig.10As shown, the terminal end of the connecting tube 206 is connected with a first sleeve 207, and an arc sleeve 210 is fixedly installed at one end of the first sleeve 207, and a sterilization tube 208 extending outward is arranged at the other end of the first sleeve 207, and one end of the sterilization tube 208 extending out of the first sleeve 207 is made of rubber, and the sterilization tube 208 contacts the outer side of the test tube 303, and a sterilization nozzle 209 is penetrated at one end of the sterilization tube 208, and one end of the arc sleeve 210 is fixedly connected to the bottom of the pressure stabilizing box 205, and the other end of the arc sleeve 210 is sleeved with a transmission tube 211 extending outward, and the transmission tube 211 is fixedly connected to the bottom of the rotating plate 302, and the rotation centers of the transmission tube 211 and the arc sleeve 210 are located on the rotation axis of the rotating plate 302, and a reset spring 212 is fixedly installed at the other end of the transmission tube 211, and the transmission tube 211 is connected to the inner wall of the arc sleeve 210 through the reset spring 212; By setting the sterilization tube 208 in contact with the outside of the test tube 303 and combining with the rotating plate 302, the test tube 303 is clamped to prevent it from falling off during rotation. At the same time, the sterilization tube 208 is provided in conjunction with the sterilization nozzle 209 to facilitate the spraying of the sterilizer, thereby achieving a sterilization effect. The connecting box 203 draws gas into the first sleeve 207 and the arc sleeve 210 through the connecting tube 206, and then the transmission of the sterilization tube 208 and the transmission tube 211 is achieved by changing the air pressure inside the first sleeve 207 and the arc sleeve 210. When the first sleeve 207 moves, it is convenient to adjust the sterilization position, and the transmission tube 211 facilitates the rotation of the rotating plate 302.

[0033] Example 3, further optimizes the brown adipocyte in vitro culture device provided in Example 1 or 2, specifically, Fig.14 As shown, the in vitro culture method specifically comprises the following steps: S1: Cell acquisition: The acquired adipose tissue stem cells are placed inside the test tube 303, and the driving motor 201 is started. The main gear 202 installed at the output end drives the transmission disc 204 engaged therewith to rotate, thereby driving the mounting plate 301 and the rotating plate 302 to rotate through the connection box 203 and the voltage stabilizing box 205, so as to centrifuge the cells inside the test tube 303, and then separate the precursor cells; S2: Cell expansion: The separated precursor cells are transferred to the interior of the culture dish 304, and an appropriate amount of culture medium can be repeatedly added to the interior of the culture dish 304 through the feeding tube 315 and the inner tank 310 to perform cell expansion culture.

[0034] Further, such as Figure 4 As shown, the following steps are also included: S3: Cell differentiation: Add culture medium containing insulin, T3, and norepinephrine to induce precursor cells to differentiate into brown adipocytes.

[0035] Further, such as Figure 4 As shown, the following steps are also included: S4: Cell maturation: Add vinblastine and dexamethasone to the culture medium and culture the brown adipocytes to promote cell maturation and obtain functional brown adipocytes.

[0036] Further, such as Figure 4 As shown, the following steps are also included: S5: Cell quality detection: The metabolic function of cultured brown adipocytes was verified by UCP1 expression and oxygen consumption rate measurement.

[0037] The use process of the brown adipocyte in vitro culture device provided by the present invention is as follows: Before use, firstly, the sterilizing agent is sprayed into the interior of the frame 1 through the sterilizing tube 208 in cooperation with the sterilizing nozzle 209 installed at one end thereof, so as to sterilize the inner cavity of the frame 1 until the sterilization is completed; At this time, the obtained fat tissue or stem cells from other sources are placed inside the test tube 303, and the test tube 303 is passed through the rotating plate 302, and the test tube 303 is clamped by the rotating plate 302. When the test tube 303 is placed, the test tube 303 passes through the groove opened on the side wall of the pressure stabilizing box 205. At this time, the air pump 213 is started, and the funnel 214 set at one end of the air pump 213 and the transmission disk 204 continuously transport gas to the connection box 203 and the pressure stabilizing box 205, so that the air pressure inside the connection box 203 and the pressure stabilizing box 205 is continuously increased, until the pressurization is completed, and the air pump 213 is stopped; At this time, gas is delivered to the inner cavity of the first sleeve 207 through the connecting tube 206, and the sterilization tube 208 is pushed by the gas to move until the sterilization tube 208 contacts the side wall of the test tube 303 and squeezes it; At this time, the driving motor 201 is started, and the main gear 202 installed at its output end drives the transmission disc 204 meshing with it to rotate, thereby driving the mounting plate 301 and the rotating plate 302 to rotate through the connection box 203 and the voltage stabilizing box 205, so as to centrifuge the cells inside the test tube 303, and then separate the precursor cells. After the centrifugation is completed, the driving motor 201 is stopped, and the staff takes out the test tube 303. After the test tube 303 is taken out, since the rotating plate 302 is connected to the mounting plate 301 through the torsion spring, the mounting plate 301 returns to its original position under the action of the torsion spring, and the rotating plate 302 returns to its original position. The initial position is tilted downward, and its inner wall is in contact with the side wall of the pressure stabilizing box 205. As the mounting plate 301 is reset, the pull rope 312 installed on the top thereof is pulled to pull the first connecting block 311 and the slider 309 upward, and the second sleeve 306 and the cover plate 305 slidably connected therewith are driven to rise through the slider 309. At this time, the extrusion spring 308 is in a stretched state, and the inner groove 310 is connected with the feeding tube 315, which is to transfer the separated precursor cells to the inside of the culture dish 304, and the appropriate amount of culture medium can be repeatedly added to the inside of the culture dish 304 through the feeding tube 315 and the inner groove 310; At this time, gas is delivered to the inner cavity of the arc sleeve 210 through the pressure stabilizing box 205, the connection box 203 and the connection tube 206. At this time, the reset spring 212 is cooperated, and the transmission tube 211 is pushed to move by the gas, thereby pushing the rotating plate 302 and the test tube 303 to rotate. The test tube 303 continuously tilts upward, thereby driving the rotating plate 302 to tilt upward. At this time, the cover plate 305 moves in the opposite direction and extends to the inside of the culture dish 304, and the cover plate 305 is sleeved through the culture dish 304; By continuously tilting the rotating plate 302, the pull rope 312 is continuously loosened, and the slider 309 moves downward under the action of the squeezing spring 308 to seal the cover plate 305 and the second sleeve 306. At this time, the ceiling induction operation is performed through the culture dish 304 to induce the precursor cells to differentiate into brown adipocytes until the differentiation is completed. During the differentiation process, the driving motor 201 can also be started to drive the voltage stabilizing box 205 to move slowly, thereby shaking the culture dish 304 to rotate, so that the culture is more uniform, and a partition can be set inside the culture dish 304 to achieve classified culture; At this time, the differentiated brown adipocytes are cultured in a culture medium containing vinblastine and dexamethasone to promote cell maturation and obtain functional brown adipocytes; at the same time, the metabolic function of the cultured brown adipocytes is verified by the UCP1 expression and oxygen consumption rate (OCR) measurement equipment inside the frame 1.

[0038] In the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", "fixed" and the like should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral one; it can be a mechanical connection, an electrical connection, or communication with each other; it can be a direct connection, or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements, unless otherwise clearly defined. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0039] Obviously, the embodiments described above are only some embodiments of the present invention, rather than all embodiments. The preferred embodiments of the present invention are given in the accompanying drawings, but they do not limit the patent scope of the present invention. The present invention can be implemented in many different forms. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive. Although the present invention has been described in detail with reference to the aforementioned embodiments, for those skilled in the art, it is still possible to modify the technical solutions recorded in the aforementioned specific embodiments, or to perform equivalent replacements for some of the technical features therein. Any equivalent structure made using the contents of the specification and drawings of the present invention, directly or indirectly used in other related technical fields, is also within the scope of patent protection of the present invention.

Claims

1. A brown adipocyte in vitro culture device, characterized in that: It comprises a frame (1), wherein an amplification component (3) for cell amplification and a separation component (2) for cell acquisition are arranged in upper and lower intervals inside the frame (1); The amplification component (3) comprises a mounting plate (301) arranged in the inner cavity of the frame (1), and a rotating plate (302) is provided in a rectangular array outside the mounting plate (301), and a test tube (303) is sleeved in the middle of the rotating plate (302), and a culture dish (304) for cell culture is sleeved in the middle of the mounting plate (301), a cover plate (305) is provided on the top of the culture dish (304), and a second sleeve (306) is connected to the top of the cover plate (305).

2. The in vitro culture device for brown adipocytes according to claim 1, characterized in that: A mounting frame is sleeved in the middle of the outer side of the second sleeve (306), and the bottom of the mounting frame is fixedly connected to the top of the mounting plate (301); a mounting pad (307) is fixedly installed at the bottom of the inner cavity of the second sleeve (306), and the mounting pad (307) is annular; a compression spring (308) is fixedly installed on the top of the mounting pad (307), and a slider (309) is fixedly installed on the top of the compression spring (308); the second sleeve (306) and the slider (309) are movably sleeved.

3. The in vitro culture device for brown adipocytes according to claim 2, characterized in that: The inner cavity of the slider (309) is provided with an inner groove (310), and the inner groove (310) is in a right angle shape. The bottom of the inner groove (310) passes through the bottom of the slider (309), and the other end of the inner groove (310) passes through the side wall of the slider (309). The top of the side wall of the second sleeve (306) is connected to a feeding pipe (315), and the feeding pipe (315) is connected to the inner groove (310).

4. The in vitro culture device for brown adipocytes according to claim 3, characterized in that: The top of the sliding block (309) is sleeved with a first connecting block (311), and a rectangular array of pull ropes (312) is provided on the top of the first connecting block (311); a second connecting block (313) is arranged above the first connecting block (311) at intervals, and a fixing block (314) is sleeved on the top of the second connecting block (313); the top of the fixing block (314) is fixedly connected to the top of the inner cavity of the frame (1).

5. The in vitro culture device for brown adipocytes according to claim 4, characterized in that: The pull rope (312) passes through the bottom of the second connecting block (313) and extends out of the side wall thereof; the other end of the second connecting block (313) is fixedly connected to the top of the rotating plate (302).

6. The in vitro culture device for brown adipocytes according to claim 5, characterized in that: The separation assembly (2) comprises a pressure stabilizing box (205) arranged at the bottom of the culture dish (304); the top of the pressure stabilizing box (205) is fixedly connected to the mounting plate (301); the side walls of the pressure stabilizing box (205) are provided with grooves, and the pressure stabilizing box (205) is sleeved with the test tube (303) through the grooves; a connection box (203) is arranged at the bottom of the pressure stabilizing box (205), and the connection box (203) is connected to the pressure stabilizing box (205); the side wall array pipe of the connection box (203) is connected to a connection pipe (206), and the outer side of the connection pipe (206) is provided with a valve body.

7. The in vitro culture device for brown adipocytes according to claim 6, characterized in that: The bottom pipe of the connection box (203) is connected to a transmission disc (204), the transmission disc (204) is tubular, and the outer side of the bottom of the transmission disc (204) is gear-shaped, the middle of the transmission disc (204) is provided with a valve body, the bottom of the transmission disc (204) is sleeved with a funnel (214), and the bottom pipe of the funnel (214) is connected to an air pump (213).

8. The in vitro culture device for brown adipocytes according to claim 7, characterized in that: A driving motor (201) is arranged at intervals on one side of the air pump (213), and a main gear (202) is fixedly mounted on the output end of the driving motor (201), and the main gear (202) is meshed with the bottom of the transmission disc (204).

9. The in vitro culture device for brown adipocytes according to claim 8, characterized in that: The terminal end of the connecting tube (206) is connected to a first sleeve (207), and an arc-shaped sleeve (210) is fixedly installed on one end of the first sleeve (207), and a sterilization tube (208) extending outward is provided at the other end of the first sleeve (207), and one end of the sterilization tube (208) extending out of the first sleeve (207) is made of rubber, and the sterilization tube (208) is in contact with the outside of the test tube (303), and a sterilization nozzle (209) is provided through one end of the sterilization tube (208).

10. The in vitro culture device for brown adipocytes according to claim 9, characterized in that: One end of the arc-shaped sleeve (210) is fixedly connected to the bottom of the voltage-stabilizing box (205); the other end of the arc-shaped sleeve (210) is sleeved with a transmission tube (211) extending outward, and the transmission tube (211) is fixedly connected to the bottom of the rotating plate (302); the rotation centers of the transmission tube (211) and the arc-shaped sleeve (210) are located on the rotation axis of the rotating plate (302); the other end of the transmission tube (211) is fixedly mounted with a return spring (212), and the transmission tube (211) is connected to the inner wall of the arc-shaped sleeve (210) via the return spring (212).

Images