CN116590128A - Cell culture exosome collecting device and collecting method - Google Patents
Cell culture exosome collecting device and collecting method Download PDFInfo
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- CN116590128A CN116590128A CN202310468332.1A CN202310468332A CN116590128A CN 116590128 A CN116590128 A CN 116590128A CN 202310468332 A CN202310468332 A CN 202310468332A CN 116590128 A CN116590128 A CN 116590128A
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- 210000001808 exosome Anatomy 0.000 title claims abstract description 93
- 238000004113 cell culture Methods 0.000 title claims abstract description 27
- 238000000034 method Methods 0.000 title claims abstract description 11
- 238000000605 extraction Methods 0.000 claims abstract description 13
- 238000007789 sealing Methods 0.000 claims description 59
- 239000007788 liquid Substances 0.000 claims description 23
- 239000003638 chemical reducing agent Substances 0.000 claims description 10
- 230000001681 protective effect Effects 0.000 claims description 9
- 239000000284 extract Substances 0.000 claims description 6
- 230000000149 penetrating effect Effects 0.000 claims description 5
- 238000005119 centrifugation Methods 0.000 abstract description 5
- 238000012360 testing method Methods 0.000 abstract description 3
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- 210000004027 cell Anatomy 0.000 description 6
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- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/08—Flask, bottle or test tube
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/38—Caps; Covers; Plugs; Pouring means
Abstract
The invention discloses a cell culture exosome collecting device and a collecting method, which relate to the field of cell culture and comprise a collecting device main body, wherein an extraction component is arranged on the other side of the collecting device main body, a placement component is arranged in the collecting device main body, the extraction component and the placement component are mutually matched for use, when exosomes are extracted, a gear motor is started firstly to drive a piston to move, exosomes are extracted into the collecting pipe, and then a placement disc is intermittently rotated through the matching of a first rotating component for batch collection, so that the problem that when exosomes are collected, all exosomes are uniformly collected into a collecting box, and then are required to be transported into all test tubes for centrifugation after being uniformly collected is avoided, the problem that operation steps are complicated is caused, and the convenience of operation is improved, so that the working efficiency is improved.
Description
Technical Field
The invention relates to the technical field of cell culture, in particular to a cell culture exosome collecting device and a cell culture exosome collecting method.
Background
Exosomes refer to small vesicles (30-150 nm) containing complex RNAs and proteins, which nowadays are specifically disc vesicles with diameters of 40-100 nm; the exosomes can be secreted by various cells under normal and pathological states, and are mainly derived from multi-vesicle bodies formed by inward sinking of lysosome particles in the cells, and are released into extracellular matrixes after being fused with cell membranes through outer membranes of the multi-vesicle bodies; all cultured cell types secrete exosomes, and exosomes naturally occur in body fluids, including blood, saliva, urine, cerebrospinal fluid and milk.
In recent years, exosomes are considered to be extremely potential tumor diagnostic markers and good carriers for drug delivery due to their wide sources, stable structure, rich inclusion and important roles in intercellular information and substance transfer, and therefore they continue to be research hotspots in the field of national research for many years.
Such as China issued patent: CN115404153a, a cell culture exosome collecting device, including the bottom plate, still include the collection mechanism that sets up on the bottom plate surface, collection mechanism includes inlet unit, jacking subassembly, spacing subassembly, dismantles the subassembly and collects the subassembly, inlet unit sets up the upper surface of bottom plate, jacking subassembly sets up inlet unit's surface, spacing subassembly is installed the upper surface of bottom plate, and through jacking subassembly with inlet unit links, collection unit sets up spacing subassembly's surface, dismantlement subassembly sets up spacing subassembly's bottom surface; the device is beneficial to enabling the supernatant to flow into the inner collecting cup at a constant speed to collect, realizing controlling the collected amount, facilitating the collected supernatant to be taken out from the inner part of the round outer sealing cup, and improving the convenience in the use process of the device.
Because the supernatant contains some cell fragments, the cell fragments can be combined with porous microspheres to influence the purity and experimental effect of exosome extraction, the exosome in the sample is usually required to be purified by adopting porous microspheres with high binding force with phosphatidylserine, the exosome in the sample is adsorbed on the surface of the porous microspheres during purification, impurities such as other non-phosphate proteins are not adsorbed and removed, the porous microspheres adsorbed with the exosome are washed by washing liquid to remove proteins, impurities and salts, and finally the exosome with high purity is obtained by eluting with eluent.
Disclosure of Invention
The invention aims to provide a cell culture exosome collecting device and a cell culture exosome collecting method, which can solve the problems in the background art.
In order to achieve the above purpose, the present invention provides the following technical solutions: the cell culture exosome collecting device comprises a collecting device main body, wherein one side of the collecting device main body is hinged with a protective door, the other side of the collecting device main body is provided with an extraction assembly, a liquid inlet pipe is embedded in the collecting device main body, and a placement assembly is arranged in the collecting device main body;
the extraction assembly comprises a first connecting rod arranged on the other side of the collecting device main body, one end of the first connecting rod is fixedly connected with a piston, the piston is slidably mounted in the first connecting rod, the other end of the first connecting rod is fixedly connected with a first connecting plate, the first connecting plate is embedded into the inner wall of the collecting device main body and is slidably connected with the first connecting rod, a second connecting rod is slidably mounted on the inner wall of the first connecting plate, one end, far away from the second connecting rod, of the first connecting plate is provided with a first rotating assembly, the top end of the second connecting rod is fixedly provided with a second connecting plate, one end, far away from the second connecting rod, of the second connecting plate is fixedly connected with a third connecting rod, the top end of the third connecting rod is provided with a speed reducer, the top end of the speed reducer is provided with a fourth connecting rod, the top end of the fourth connecting rod is fixedly connected with a speed reducer motor, and the speed reducer is fixedly mounted in the collecting device main body;
wherein, place the subassembly including setting up the inside dish of placing of collection device main part, place the dish and rotate through coupling assembling and collection device main part and be connected, the inside of placing the dish is circumference array evenly and is provided with the collecting pipe, the collecting pipe sets up the bottom at the feed liquor pipe, and the top of one of them collecting pipe is provided with seal assembly.
Preferably, the connecting component comprises a first connecting column fixedly arranged at the bottom end of the placing tray, a mounting column is slidably arranged at the bottom end of the first connecting column, a second connecting column is slidably arranged at the bottom end of the mounting column, the second connecting column is rotatably arranged in the collecting device main body, a first spring is arranged between the mounting column and the second connecting column,
preferably, the first rotating assembly comprises a third connecting plate arranged at one end of the first connecting plate far away from the second connecting rod, one side of the third connecting plate is uniformly provided with tooth blocks, the tooth blocks are provided with two groups, one side of one group of tooth blocks is meshed with the bottom end of the second connecting column, the other side of the other group of tooth blocks is provided with a second rotating assembly,
preferably, the second rotating assembly comprises a gear arranged at one side of the other group of the tooth blocks, the tooth blocks are meshed with the gear, a conical gear set is fixedly arranged at the bottom end of the gear, a seventh connecting rod is fixedly arranged at the other side of the conical gear set, a cam is fixedly arranged at one end of the seventh connecting rod far away from the conical gear set,
preferably, each tooth block is rotationally connected with the third connecting plate through a fifth connecting rod, a torsion spring is arranged in each fifth connecting rod,
preferably, one side of each tooth block is provided with an inclined plane, and the inclined planes of the two groups of tooth blocks are opposite in position
Preferably, the sealing component comprises a sealing cover arranged at the top end of the collecting pipe, the sealing cover is slidably arranged in the accommodating barrel, the bottom end of the accommodating barrel is provided with a moving component, one side of the accommodating barrel is fixedly connected with a first lug, the first lug is arranged at the top end of the cam,
preferably, the moving assembly comprises a first lug slidably mounted in the bottom end of the accommodating barrel, the first lugs are respectively arranged on two sides of the accommodating barrel, a second spring is mounted between the two first lugs and the accommodating barrel, the bottom ends of the two first lugs are respectively provided with a second lug, and the two second lugs are respectively and fixedly mounted on two sides of the collecting pipe
Preferably, the inner wall at the top end of the placing barrel is slidably provided with a supporting plate, the top end of the supporting plate is fixedly provided with a pull rod, and the outer wall of the pull rod is sleeved with a third spring.
A cell culture exosome collection method:
s1: when the exosome is required to be collected, the position of the main body of the collecting device is firstly adjusted, and one end of the liquid inlet pipe penetrating through the inner wall of the main body of the collecting device is placed in the exosome;
s2: then, starting a speed reducing motor through an external power supply, and enabling the inside of the collecting device main body to form a negative pressure cavity through the extracting component, so that the liquid inlet pipe extracts an external body to the inside of the collecting pipe for collection;
s3: after the exosome is collected in one collecting pipe, the placing component is driven to rotate by the first rotating component, so that the collecting pipe which has collected the exosome moves to the bottom end of the sealing component, and the collecting pipe which has not collected the exosome moves to the bottom end of the liquid inlet pipe;
s4: the second rotating assembly drives the sealing assembly to move downwards to seal the opening of the collected exosome collecting pipe, and the extracting assembly works to enable the device to continuously collect and seal the exosome;
s5: after the collection is completed, the gear motor is closed, the protective door is opened, the placing plate is taken out of the collection device body through the connecting assembly, and the collection pipe is taken out for the next step.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention uses the extraction component and the placement component in a mutual matching way, when the exosome is extracted, the position of the main body of the collecting device is firstly adjusted, the liquid inlet pipe passes through one end of the inner wall of the main body of the collecting device and is placed in the exosome to be collected, then the speed reducing motor is started by an external power supply, the input end of the speed reducing motor is electrically connected with the external power supply through a lead, the output end of the speed reducing motor is fixedly connected with the fourth connecting rod, the fourth connecting rod enables the third connecting rod to rotate at a low speed through the speed reducing device, the speed reducing device is a common speed reducing box, the speed reducing device works by utilizing a big gear and a small gear, the number of the gear teeth of the input gear shaft is smaller than the number of the gear teeth of the big gear on the output shaft because the gear teeth of the input gear shaft are meshed together, the number of the gear teeth of the input gear shaft is inversely proportional to the rotation number ratio according to the tooth ratio, when the power source (such as a motor) or other transmission mechanism moves at a high speed, the output shaft is lower than the input shaft to achieve the aim of deceleration after being transmitted to the output shaft through the input gear shaft), the rotation of the third connecting rod drives the rotation of the second connecting plate, the rotation of the second connecting plate drives the second connecting rod to rotate, the second connecting rod slides in the first connecting plate and drives the first connecting plate to move in the inner wall of the collecting device main body, the first connecting plate drives the first connecting rod to move, the movement of the first connecting rod drives the piston to move, the piston can be made of rubber materials, because the piston is in sealing connection with the inner part of the collecting device main body, when the piston moves to the side far away from the collecting device main body, negative pressure is formed in the inner part of the collecting device main body, and because the liquid inlet pipe is communicated with the inner part of the collecting device main body, can make the piston remove and make the drive feed liquor pipe extract the exosome, extract the exosome to the inside of collecting pipe, the cooperation of rethread first rotating assembly makes the placing plate intermittent type rotate, thereby let the exosome collect in proper order to the inside of different collecting pipes, collect in batches, avoided when collecting the exosome, the inside of collecting the box is all unified to all exosome, and after the exosome is collected, still need come to remove cell to it centrifugation, piece and impurity, and follow-up purification operation, after the unified collection, still need transport to each test tube inside, carry out centrifugation, thereby lead to the comparatively loaded down with trivial details problem of operating procedure, thereby the convenience of operation has been improved and the work efficiency is improved.
2. According to the invention, when the collecting pipe which collects the exosome is required to be sealed, power can be transmitted to the second rotating assembly through the extracting assembly, so that the placing barrel moves up and down, when the collecting pipe which has collected the exosome moves to the bottom end of the placing barrel, the first lug moves downwards under the action of the second rotating assembly, the placing barrel moves downwards to drive the sealing cover to move, the sealing cover is arranged as a common sealing plug and consists of two circular tables, the diameter of the upper circular table is larger than that of the lower circular table, the lower circular table is in interference fit with the collecting pipe, when the sealing cover moves downwards, the opening of the collecting pipe can be embedded, meanwhile, the placing barrel moves downwards to drive the first lug to move, as the bottom end of the first lug, which is close to the sealing cover, is arranged as an inclined plane, and one side, which is far away from the collecting pipe, of the top end of the second lug is matched with the inclined plane, when the first lug moves downwards, the second lug is contacted with the second lug, the second spring is pressed by the force of the second lug, the first lug deforms to store the elastic force, the first lug slides into the interior of the placing barrel to enable the sealing cover to be contacted with the sealing cover, and the sealing cover to be separated from the interior of the placing barrel, so that the sealing cover is separated from the sealing cover, and the opening of the collecting pipe can be in the sealing cover, and the air can be exposed to the exosome can be exposed, and exposed to the air can be exposed in the air, and the air can be exposed outside in the air can be detected, and exposed to the air can be exposed to the outside is detected in the outside is exposed to the outside.
Drawings
FIG. 1 is a schematic diagram of the overall structure of the present invention;
FIG. 2 is a schematic cross-sectional view of the overall structure of the present invention;
FIG. 3 is a schematic cross-sectional view of another view of the overall structure of the present invention;
FIG. 4 is a schematic view of the structure of the connecting assembly of the present invention;
FIG. 5 is an enlarged schematic view of the structure of FIG. 2A according to the present invention;
FIG. 6 is an enlarged schematic view of the structure of FIG. 4B according to the present invention;
fig. 7 is an enlarged view of the structure of fig. 2 at C according to the present invention.
In the figure: 1. a collection device body; 2. a protective door; 3. a liquid inlet pipe; 4. a first connecting rod; 5. a piston; 6. a first connection plate; 7. a second connecting rod; 8. a second connecting plate; 9. a third connecting rod; 10. a speed reducing device; 11. a fourth connecting rod; 12. a speed reducing motor; 13. a collection pipe; 14. placing a tray; 15. a first connection post; 16. a mounting column; 17. a second connection post; 18. a first spring; 19. a third connecting plate; 20. tooth blocks; 21. a fifth connecting rod; 22. a sixth connecting rod; 23. a gear; 24. a conical gear set; 25. a seventh connecting rod; 26. a cam; 27. sealing cover; 28. placing a barrel; 29. a first bump; 30. a second spring; 31. a second bump; 32. a support plate; 33. a pull rod; 34. a third spring; 35. and a torsion spring.
Description of the embodiments
The following description of the embodiments of the present invention will be made clearly and fully with reference to the accompanying drawings, in which it is evident that the embodiments described are only some, but not all embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Examples
Referring to fig. 1-7, a cell culture exosome collecting device and a collecting method thereof are shown in the drawings, comprising a collecting device main body 1, wherein a protective door 2 is hinged on one side of the collecting device main body 1, an extraction component is arranged on the other side of the collecting device main body 1, a liquid inlet pipe 3 is embedded in the collecting device main body 1, and a placement component is arranged in the collecting device main body 1.
Wherein the extraction assembly comprises a first connecting rod 4 arranged at the other side of the collecting device main body 1, one end of the first connecting rod 4 is fixedly connected with a piston 5, the piston 5 is slidably mounted in the first connecting rod 4, the other end of the first connecting rod 4 is fixedly connected with a first connecting plate 6, the first connecting plate 6 is embedded in the inner wall of the collecting device main body 1 and is slidably connected with the first connecting rod 7, the inner wall of the first connecting plate 6 is slidably mounted with a second connecting rod 7, one end of the first connecting plate 6 far away from the second connecting rod 7 is provided with a first rotating assembly, the top end of the second connecting rod 7 is fixedly mounted with a second connecting plate 8, one end of the second connecting plate 8 far away from the second connecting rod 7 is fixedly connected with a third connecting rod 9, the top end of the third connecting rod 9 is provided with a speed reducer 10, the top end of the speed reducer 10 is provided with a fourth connecting rod 11, the top end of the fourth connecting rod 11 is fixedly connected with a speed reducer 12, the speed reducer 12 is fixedly mounted in the collecting device main body 1,
wherein, place the subassembly and including setting up the setting at the inside dish 14 of placing of collection device main part 1, place the dish 14 and rotate through coupling assembling and collection device main part 1 and be connected, the inside of placing the dish 14 is circumference array and evenly is provided with collecting pipe 13, and collecting pipe 13 sets up the bottom at feed liquor pipe 3, and the top of one of them collecting pipe 13 is provided with seal assembly.
When the exosome is extracted, the position of the collecting device main body 1 is adjusted, one end of the liquid inlet pipe 3 penetrating through the inner wall of the collecting device main body 1 is placed in the exosome to be collected, then the speed reducing motor 12 is started through an external power supply, the input end of the speed reducing motor 12 is electrically connected with the external power supply through a wire, the output end of the speed reducing motor 12 is fixedly connected with the fourth connecting rod 11, the fourth connecting rod 11 enables the third connecting rod 9 to rotate at a low speed through the speed reducing device 10, the speed reducing device 10 is a common speed reducing box and works by utilizing the big and small gears 23, as the gear teeth of the input gear 23 shaft are meshed with the big gear 23 on the output shaft, the gear teeth of the input gear 23 shaft are less than the gear teeth of the big gear 23 on the output shaft, according to the inverse proportion of the number of teeth and the revolution, when the high-speed motion of a power source (such as a motor) or other transmission mechanisms is transmitted to the output shaft through the input gear 23, the output shaft obtains low-speed motion lower than the input shaft, so as to achieve the aim of reducing speed, the rotation of the third connecting rod 9 drives the rotation of the second connecting plate 8, the rotation of the second connecting plate 8 drives the second connecting rod 7 to rotate, the second connecting rod 7 slides in the first connecting plate 6 and drives the first connecting plate 6 to move in the inner wall of the collecting device main body 1, the first connecting plate 6 drives the first connecting rod 4 to move, the movement of the first connecting rod 4 drives the piston 5 to move, the piston 5 can be made of rubber materials, because the piston 5 is in sealing connection with the inner part of the collecting device main body 1, when the piston 5 moves to the side far away from the collecting device main body 1, negative pressure is formed in the inner part of the collecting device main body 1, the liquid inlet pipe 3 is communicated with the inside of the collecting device main body 1, so that the piston 5 can move to drive the liquid inlet pipe 3 to extract exosomes, the exosomes are extracted to the inside of the collecting pipe 13, the placing plate 14 is intermittently rotated through the cooperation of the first rotating component, the exosomes are sequentially collected to the inside of different collecting pipes 13, batch collection is carried out, the problem that when the exosomes are collected, all exosomes are uniformly collected to the inside of the collecting box is avoided, after the exosomes are collected, centrifugation is needed to be carried out to remove cells, fragments and impurities, and subsequent purification operation is carried out, after the exosomes are uniformly collected, the exosomes are also needed to be transferred to the inside of each test tube, and centrifugation is carried out, so that the problem that operation steps are complicated is caused, and the convenience of operation is improved, and the working efficiency is improved.
The connecting component comprises a first connecting column 15 fixedly arranged at the bottom end of the placement disc 14, a mounting column 16 is slidably arranged at the bottom end of the first connecting column 15, a second connecting column 17 is slidably arranged at the bottom end of the mounting column 16, the second connecting column 17 is rotatably arranged in the collecting device main body 1, a first spring 18 is arranged between the mounting column 16 and the second connecting column 17, when the collecting tube 13 needs to be taken out of the collecting device main body 1 through the connecting component, the protective door 2 is firstly opened, the mounting column 16 is slidably arranged on the inner wall of the collecting device main body 1, the mounting column 16 can be manually pressed downwards by a worker, the mounting column 16 is made to extrude the first spring 18 to deform to store elastic potential energy, meanwhile, the mounting column 16 is downwardly slid on the outer wall of the second connecting column 17, the inner wall of the mounting column 16 is separated from the outer wall of the first connecting column 15, then the placement disc 14 is outwardly moved, and the collecting tube 13 can be directly and upwardly slid and taken out due to the fact that the placement disc 14 is slidably connected with the collecting tube 13, and the collecting tube 13 is conveniently taken out.
The first rotating assembly comprises a third connecting plate 19 arranged at one end, far away from the second connecting rod 7, of the first connecting plate 6, tooth blocks 20 are uniformly arranged on one side of the third connecting plate 19, two groups of tooth blocks 20 are arranged, one side of one group of tooth blocks 20 is meshed with the bottom end of the second connecting column 17, the second rotating assembly is arranged on one side of the other group of tooth blocks 20, when the exosomes are required to be sequentially collected into each collecting pipe 13 through the first rotating assembly, the first connecting plate 6 can be driven to move by the first connecting plate 6 through the operation of the extracting assembly, the movement of the third connecting plate 19 can drive the movement of the tooth blocks 20, the movement of the first group of tooth blocks 20 drives the rotation of the connecting assembly, the number of the first group of tooth blocks 20 can be set to be one N times of the number of teeth of the outer wall of the second connecting column 17, and N is the number of the collecting pipes 13, so that the collecting pipes 13 are driven to move when the placing plate 14 rotates each time, and the exosomes are sequentially collected.
The second rotating assembly comprises a gear 23 arranged on one side of the other group of tooth blocks 20, the tooth blocks 20 are meshed with the gear 23, a conical gear set 24 is fixedly arranged at the bottom end of the gear 23, a seventh connecting rod 25 is fixedly arranged at the other side of the conical gear set 24, a cam 26 is fixedly arranged at one end of the seventh connecting rod 25 far away from the conical gear set 24, the second rotating assembly can be matched with the sealing assembly for use, after the first rotating assembly drives the collecting assembly to rotate, when the second group of tooth blocks 20 move, the tooth blocks 20 drive the gear 23 to rotate, the gear 23 rotates to drive the sixth connecting rod 22 to rotate, the rotation of the sixth connecting rod 22 drives the conical gear set 24 to rotate, the rotation of the conical gear set 24 drives the seventh connecting rod 25 to drive the cam 26 to be matched with the tooth number of the gear 23, the gear 23 can drive the seventh connecting rod 25 to rotate for one circle each time, and as the cam 26 is arranged into an irregular shape, and when the first rotating assembly is in an initial state, the highest point of the cam 26 firstly drives the sealing assembly to move downwards, and then the sealing assembly is driven to move upwards by the sealing assembly.
Each tooth block 20 is rotationally connected with the third connecting plate 19 through a fifth connecting rod 21, a torsion spring 35 is installed in each fifth connecting rod 21, through the torsion spring 35, when the third connecting plate 19 moves, the torsion spring 35 is extruded to generate deformation to store elastic potential energy when the first group of tooth blocks 20 or the second group of tooth blocks 20 are contacted with the tooth blocks 20 of the second connecting column 17 or the tooth blocks 20 of the gear 23, and when the first group of tooth blocks 20 or the second group of tooth blocks 20 are not contacted with the tooth blocks 20 of the second connecting column 17 or the tooth blocks 20 of the gear 23, the torsion spring 35 releases the elastic potential energy, and the tooth blocks 20 are restored to the original positions under the action of the torsion spring 35.
One side of each tooth block 20 is set to be an inclined plane, the inclined planes of the two groups of tooth blocks 20 are opposite in position, when the third connecting plate 19 moves, the movement of the third connecting plate 19 drives the first group of tooth blocks 20 and the second group of tooth blocks 20 to move simultaneously, the inclined plane of the first group of tooth blocks 20 is the direction in which the third connecting plate 19 moves away from the second connecting column 17, when the third connecting plate 19 slides to one side of the second connecting column 17, the first group of tooth blocks 20 can drive the second connecting column 17 to rotate, so that the placing assembly is driven to rotate, meanwhile, because the inclined plane of the second group of tooth blocks 20 is the direction in which the third connecting plate 19 moves close to the second connecting column 17, the second group of tooth blocks 20 does not drive the sixth connecting rod 22 to rotate, when the third connecting plate 19 slides to one side away from the second connecting column 17, the second group of tooth blocks 20 drive the gears 23 to rotate, thereby providing power for the sealing assembly, the movement interference of the placing assembly and the movement stability of the sealing assembly in the movement process of the third connecting plate 19 is prevented.
A cell culture exosome collection method:
s1: when the exosome of cell culture needs to be collected, firstly, the position of the collecting device main body 1 is adjusted, and one end of the liquid inlet pipe 3 penetrating through the inner wall of the collecting device main body 1 is placed in the exosome;
s2: then the speed reducing motor 12 is started by an external power supply, and a negative pressure cavity is formed in the collecting device main body 1 by the extraction component, so that the liquid inlet pipe 3 extracts an exosome to the inside of the collecting pipe 13 for collection;
s3: after the exosome is collected in one of the collecting pipes 13, the placing component is driven to rotate by the first rotating component, so that the collecting pipe 13 which has collected the exosome moves to the bottom end of the sealing component, and the collecting pipe 13 which has not collected the exosome moves to the bottom end of the liquid inlet pipe 3;
s4: the second rotating component drives the sealing component to move downwards to seal the opening of the collected exosome collecting pipe 13, and the device continuously collects and seals the exosome through the operation of the extracting component;
s5: after the collection is completed, the gear motor 12 is closed, the protective door 2 is opened, the placing tray 14 is taken out of the collection device body 1 through the connecting assembly, and the collection tube 13 is taken out for the next step.
Examples
Referring to fig. 1-7, this embodiment further illustrates example 1, the seal assembly includes the sealed lid 27 that sets up on the top of collecting pipe 13, sealed lid 27 slidable mounting is in the inside of placing the bucket 28, the bottom of placing the bucket 28 is provided with the removal subassembly, one side fixedly connected with first lug 29 of placing the bucket 28, first lug 29 sets up on the top of cam 26, through the seal assembly that sets up, when need seal collecting pipe 13 to the exosome, can provide power transmission to the second rotating assembly through the extraction subassembly, make and place the bucket 28 reciprocate, when the collecting pipe 13 that has collected the exosome moves to the bottom of placing the bucket 28, first lug 29 moves down under the effect of second rotating assembly, place the bucket 28 and move down and drive sealed lid 27 and remove, sealed lid 27 sets up to be common sealing plug, by two round platforms, the round platform diameter below is bigger than, the round platform diameter below cooperates with collecting pipe 13, can imbeds the opening part of collecting pipe 13 when sealed lid 27 moves down, seal collecting box's opening part when need to collect the exosome, can not influence the exosome, the exosome can be exposed to the air with the exosome, the external air can be influenced by the time, if the external air can be exposed in the time, the external air can be guaranteed, the effect is exposed to the external air can be guaranteed, the external air can be influenced.
The movable assembly comprises a first lug 29 which is slidably arranged inside the bottom end of a placing barrel 28, the first lug 29 is respectively arranged on two sides of the placing barrel 28, a second spring 30 is arranged between the two first lugs 29 and the placing barrel 28, second lugs 31 are respectively arranged at the bottom ends of the two first lugs 29, the two second lugs 31 are respectively fixedly arranged on two sides of a collecting pipe 13, when the sealing assembly moves downwards, the placing barrel 28 moves downwards to drive the first lugs 29 to move through the arranged movable assembly, as the bottom end of the first lugs 29 close to a sealing cover 27 is arranged to be an inclined surface, one side, far away from the collecting pipe 13, of the top end of the second lugs 31 is arranged to be an inclined surface, the two are matched, when the first lugs 29 move downwards to be in contact with the second lugs 31, the second springs 30 are pressed by the force of the second lugs 31 to deform and store elastic potential energy, the first lugs 29 slide into the inside of the placing barrel 28 to enable the first lugs 29 to be separated from the sealing cover 27, the sealing cover 27 slides out of the inside of the placing barrel 28 to enable the sealing cover 27 to seal the collecting pipe 13, and when the sealing cover 27 slides downwards to slide out of the inside the placing barrel 28 to be clamped by the second springs 30, and the sealing cover 27 is prevented from sliding downwards, and the sealing cover 27 is guaranteed to slide out of the sealing cover 27 after the sealing cover 27 slides downwards from the inside the placing barrel 28 to be clamped by the second springs.
The inner wall on the top end of the placing barrel 28 is slidably provided with a supporting plate 32, the top end of the supporting plate 32 is fixedly provided with a pull rod 33, the outer wall of the pull rod 33 is sleeved with a third spring 34, after the bottom end of the sealing cover 27 is embedded into the collecting pipe 13, the number of the sealing covers 27 in the placing barrel 28 is reduced, the third spring 34 drives the supporting plate 32 to downwardly extrude the sealing covers 27, so that the sealing covers 27 tightly press the collecting pipe 13, and when the sealing assembly seals the collecting pipe 13, the sealing effect on the collecting pipe 13 is improved due to the fact that the sealing is not tight enough.
Working principle: when the device is required to be used for collecting the cell culture exosome, firstly, the position of the collecting device main body 1 is adjusted, and one end of the liquid inlet pipe 3 penetrating through the inner wall of the collecting device main body 1 is placed in the exosome;
then, a speed reducing motor 12 is started through an external power supply, the speed reducing motor 12 is started to drive the fourth connecting rod 11 to rotate, the fourth connecting rod 11 drives the third connecting rod 9 to rotate through a speed reducing device 10, the third connecting rod 9 rotates to drive the second connecting plate 8 to rotate, the second connecting plate 8 rotates to drive the second connecting rod 7 to rotate, the second connecting rod 7 slides in the inner wall of the first connecting plate 6 to drive the first connecting plate 6 to reciprocate in the inner wall of the collecting device main body 1, when the first connecting plate 6 moves to the side far away from the collecting device main body 1, the movement of the first connecting plate 6 drives the movement of the first connecting rod 4, the movement of the first connecting rod 4 drives the piston 5 to enable the inside of the collecting device main body 1 to form a negative pressure chamber, external secretion is extracted through the liquid inlet pipe 3, and the external secretion enters the inner wall of the collecting pipe 13 through the liquid inlet pipe 3 to be collected;
after the exosomes are collected into the collecting pipe 13, when the first connecting plate 6 moves to the side close to the collecting device main body 1, the movement of the first connecting plate 6 drives the movement of the third connecting plate 19, the movement of the third connecting plate 19 drives the movement of the tooth block 20, the first group of tooth blocks 20 drive the second connecting column 17 to rotate, the rotation of the second connecting column 17 drives the rotation of the mounting column 16, the rotation of the mounting column 16 drives the rotation of the first connecting column 15, the rotation of the first connecting column 15 drives the placing tray 14, the rotation of the placing tray 14 drives the rotation of the collecting pipe 13, and the collecting pipe 13 which has collected the exosomes rotates to the bottom end of the placing barrel 28;
when the collecting pipe 13 of the exosome has been collected and rotated to the bottom end of the placing barrel 28, the first connecting plate 6 moves to the side far away from the collecting device main body 1, and similarly, the movement of the first connecting plate 6 drives the movement of the third connecting plate 19, the movement of the third connecting plate 19 drives the movement of the tooth block 20, the first group of tooth blocks 20 does not drive the second connecting post 17 to rotate, the second group of tooth blocks 20 drives the gear 23 to rotate, the rotation of the gear 23 drives the rotation of the sixth connecting rod 22, the rotation of the sixth connecting rod 22 drives the rotation of the conical gear set 24, the rotation of the conical gear set 24 drives the rotation of the seventh connecting rod 25, the rotation of the seventh connecting rod 25 drives the rotation of the cam 26, the rotation of the cam 26 drives the first bump 29 to move up and down, the movement of the first protruding block 29 drives the movement of the placing barrel 28, the downward movement of the placing barrel 28 drives the sealing cover 27 to move downwards, the bottom end of the sealing cover 27 is embedded into the inner wall of the collecting pipe 13, when the second protruding block 31 is contacted with the first protruding block 29, the second protruding block 31 forces the first protruding block 29 to press the second spring 30 to slide in the inner wall of the placing barrel 28, the first protruding block 29 is removed from the bottom end of the sealing cover 27, meanwhile, the third spring 34 presses the supporting plate 32 to move downwards, the supporting plate 32 presses the sealing cover 27 to move downwards, the sealing cover 27 is clamped with the collecting pipe 13, the collecting pipe 13 is sealed, then the cam 26 drives the placing barrel 28 to move upwards to return to the original position, the device continuously extracts exosome, rotates and seals, and the reducing motor 12 is closed after collection is completed;
when the collecting pipe 13 needs to be taken out from the inside of the collecting device main body 1, the protective door 2 is opened, the mounting column 16 is manually pressed downwards, the mounting column 16 moves downwards to press the first spring 18, the mounting column 16 slides downwards on the outer wall of the second connecting column 17, the first connecting column 15 is separated from the inner wall of the mounting column 16, then the placing disc 14 is taken out, and the collecting pipe 13 is sequentially taken down, so that the subsequent operation is facilitated;
when the sealing cover 27 needs to be added, the cover at the top of the collecting device main body 1 can be opened, then the pull rod 33 is pulled upwards, the movement of the pull rod 33 drives the supporting plate 32 to press the third spring 34 to move upwards, a certain gap is formed between the supporting plate 32 and the opening of the placing barrel 28, and the sealing cover 27 is placed in the gap between the supporting plate 32 and the opening of the placing barrel 28 for adding.
It is noted that relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process-method-article or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process-method-article or apparatus.
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes-modifications-substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. A cell culture exosome collection device, characterized in that: comprises a collecting device main body (1), wherein one side of the collecting device main body (1) is hinged with a protective door (2), the other side of the collecting device main body (1) is provided with an extraction component, a liquid inlet pipe (3) is embedded in the collecting device main body (1), a placement component is arranged in the collecting device main body (1),
wherein the extraction assembly comprises a first connecting rod (4) arranged on the other side of the collecting device main body (1), one end of the first connecting rod (4) is fixedly connected with a piston (5), the piston (5) is slidably mounted in the first connecting rod (4), the other end of the first connecting rod (4) is fixedly connected with a first connecting plate (6), the first connecting plate (6) is embedded into the inner wall of the collecting device main body (1) and is slidably connected with a second connecting rod (7), one end of the first connecting plate (6) far away from the second connecting rod (7) is provided with a first rotating assembly, one end of the second connecting rod (7) is fixedly provided with a second connecting plate (8), one end of the second connecting plate (8) far away from the second connecting rod (7) is fixedly connected with a third connecting rod (9), the top end of the third connecting rod (9) is provided with a speed reducer (10), the top end of the speed reducer (10) is provided with a fourth connecting rod (11), one end of the fourth connecting rod (11) is slidably mounted with a motor (12) far away from the second connecting rod (7), the motor (12) is fixedly connected with the motor (12) in the collecting device (1),
the collecting device comprises a collecting device body (1), and is characterized in that the placing component comprises a placing disc (14) arranged inside the collecting device body (1), the placing disc (14) is rotationally connected with the collecting device body (1) through a connecting component, collecting tubes (13) are uniformly arranged inside the placing disc (14) in a circumferential array, the collecting tubes (13) are arranged at the bottom end of the liquid inlet tube (3), and a sealing component is arranged at the top end of one collecting tube (13).
2. The cell culture exosome collection apparatus according to claim 1, wherein: the connecting assembly comprises a first connecting column (15) fixedly arranged at the bottom end of the placing disc (14), a mounting column (16) is slidably arranged at the bottom end of the first connecting column (15), a second connecting column (17) is slidably arranged at the bottom end of the mounting column (16), the second connecting column (17) is rotatably arranged in the collecting device main body (1), and a first spring (18) is arranged between the mounting column (16) and the second connecting column (17).
3. The cell culture exosome collection apparatus according to claim 1, wherein: the first rotating assembly comprises a third connecting plate (19) arranged at one end, far away from the second connecting rod (7), of the first connecting plate (6), tooth blocks (20) are uniformly arranged on one side of the third connecting plate (19), two groups of tooth blocks (20) are arranged, one group of tooth blocks is meshed with the bottom end of the second connecting column (17), and the other group of tooth blocks is provided with a second rotating assembly.
4. A cell culture exosome collection apparatus according to claim 3, wherein: the second rotating assembly comprises a gear (23) arranged on one side of the other group of the tooth blocks (20), the tooth blocks (20) are meshed with the gear (23), a conical gear set (24) is fixedly arranged at the bottom end of the gear (23), a seventh connecting rod (25) is fixedly arranged at the other side of the conical gear set (24), and a cam (26) is fixedly arranged at one end, far away from the conical gear set (24), of the seventh connecting rod (25).
5. The cell culture exosome collection apparatus according to claim 4, wherein: each tooth block (20) is rotationally connected with a third connecting plate (19) through a fifth connecting rod (21), and a torsion spring (35) is arranged in each fifth connecting rod (21).
6. The cell culture exosome collection apparatus according to claim 4, wherein: one side of each tooth block (20) is provided with an inclined plane, and the inclined planes of the two groups of tooth blocks (20) are opposite in position.
7. The cell culture exosome collection apparatus according to claim 1, wherein: the sealing assembly comprises a sealing cover (27) arranged at the top end of the collecting pipe (13), the sealing cover (27) is slidably arranged in a placing barrel (28), a moving assembly is arranged at the bottom end of the placing barrel (28), a first lug (29) is fixedly connected to one side of the placing barrel (28), and the first lug (29) is arranged at the top end of the cam (26).
8. The cell culture exosome collection apparatus according to claim 7, wherein: the movable assembly comprises first protruding blocks (29) which are slidably mounted in the bottom end of the placement barrel (28), the first protruding blocks (29) are respectively arranged on two sides of the placement barrel (28), second springs (30) are mounted between the two first protruding blocks (29) and the placement barrel (28), second protruding blocks (31) are respectively arranged at the bottom ends of the two first protruding blocks (29), and the two second protruding blocks (31) are respectively fixedly mounted on two sides of the collecting pipe (13).
9. The cell culture exosome collection apparatus according to claim 8, wherein: the inner wall at the top end of the placing barrel (28) is slidably provided with a supporting plate (32), a pull rod (33) is fixedly arranged at the top end of the supporting plate (32), and a third spring (34) is sleeved on the outer wall of the pull rod (33).
10. The method for collecting cell culture exosomes according to claim 1, wherein:
s1: when the exosome is required to be collected in cell culture, firstly, the position of the collecting device main body (1) is adjusted, and one end of the liquid inlet pipe (3) penetrating through the inner wall of the collecting device main body (1) is placed in the exosome;
s2: then, a speed reducing motor (12) is started through an external power supply, and a negative pressure cavity is formed in the collecting device main body (1) through an extraction component, so that the liquid inlet pipe (3) extracts an external body to the inside of the collecting pipe (13) for collection;
s3: after the exosome is collected in one collecting pipe (13), the placing assembly is driven to rotate by the first rotating assembly, so that the collecting pipe (13) which is already collected with the exosome moves to the bottom end of the sealing assembly, and the collecting pipe (13) which is not collected with the exosome moves to the bottom end of the liquid inlet pipe (3);
s4: the second rotating assembly drives the sealing assembly to move downwards to seal the opening of the collected exosome collecting pipe (13), and the device continuously collects and seals the exosome through the operation of the extracting assembly;
s5: after the collection is completed, the gear motor (12) is closed, the protective door (2) is opened, the placing plate (14) is taken out of the collection device main body (1) through the connecting assembly, and the collection pipe (13) is taken out for the next step.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310468332.1A CN116590128A (en) | 2023-04-27 | 2023-04-27 | Cell culture exosome collecting device and collecting method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310468332.1A CN116590128A (en) | 2023-04-27 | 2023-04-27 | Cell culture exosome collecting device and collecting method |
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| CN116590128A true CN116590128A (en) | 2023-08-15 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202310468332.1A Withdrawn CN116590128A (en) | 2023-04-27 | 2023-04-27 | Cell culture exosome collecting device and collecting method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117402736A (en) * | 2023-12-12 | 2024-01-16 | 济南市人民医院 | Bioreactor for secretion, separation and collection of exosomes |
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2023
- 2023-04-27 CN CN202310468332.1A patent/CN116590128A/en not_active Withdrawn
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117402736A (en) * | 2023-12-12 | 2024-01-16 | 济南市人民医院 | Bioreactor for secretion, separation and collection of exosomes |
| CN117402736B (en) * | 2023-12-12 | 2024-03-08 | 济南市人民医院 | Bioreactor for secretion, separation and collection of exosomes |
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Application publication date: 20230815 |