CN113941454B - Centrifugal device and centrifugal method for collecting Cyr61 target spot conditioned medium - Google Patents
Centrifugal device and centrifugal method for collecting Cyr61 target spot conditioned medium Download PDFInfo
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- CN113941454B CN113941454B CN202111151536.XA CN202111151536A CN113941454B CN 113941454 B CN113941454 B CN 113941454B CN 202111151536 A CN202111151536 A CN 202111151536A CN 113941454 B CN113941454 B CN 113941454B
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
- B04B—CENTRIFUGES
- B04B5/00—Other centrifuges
- B04B5/04—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers
- B04B5/0407—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers for liquids contained in receptacles
- B04B5/0414—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers for liquids contained in receptacles comprising test tubes
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
- B04B—CENTRIFUGES
- B04B5/00—Other centrifuges
- B04B5/04—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers
- B04B5/0442—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
- B04B—CENTRIFUGES
- B04B5/00—Other centrifuges
- B04B5/04—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers
- B04B5/0442—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
- B04B2005/0478—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation with filters in the separation chamber
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Abstract
The invention discloses a centrifugal device and a centrifugal method for collecting Cyr61 target point conditioned medium, wherein the centrifugal device for collecting the Cyr61 target point conditioned medium comprises a supporting component, a centrifugal component, a filtering component and a liquid taking component, the supporting component comprises a base, a shell and a supporting frame, the centrifugal component comprises a driving motor, a connecting shaft, a supporting arm and a fixing piece, the filtering component comprises an electric push rod, a connecting frame and a plurality of filtering pieces, and the liquid taking component comprises a suction pump, a main pipe, a plurality of branch pipes and a plurality of suction heads. According to the invention, the culture solution is simply and conveniently separated, and meanwhile, after the separation is finished, the culture solution is filtered again and the supernatant is pumped, so that a good separation effect is achieved, and the quality of the supernatant is ensured.
Description
Technical Field
The invention relates to the technical field of centrifugation, in particular to a centrifugation device and a centrifugation method for collecting Cyr61 target point conditioned medium.
Background
Pancreatic cancer liver metastasis prognosis is poor, effective treatment means are lacking so far, and searching for a new treatment target would have important clinical value. The previous projects of the subject group demonstrated that clearing pancreatic products down-regulates Cyr61 to regulate EMT and inhibit pancreatic cancer liver metastasis, and we found in subsequent studies that clearing pancreatic products could inhibit tumor-associated macrophage (TAM) infiltration and IL-6 secretion. This therefore proposes the hypothesis: the inhibition of pancreatic cancer liver metastasis by reducing Cyr61 regulation of EMT by pancreatic volume clearance depends on the involvement of tumor microenvironment TAM, cyr61 possibly promoting the growth and metastasis of pancreatic cancer cells by regulating the tumor-associated macrophage-pancreatic cancer cell interaction feedback loop. In order to verify the hypothesis, the action mechanism of Cyr61 for regulating tumor-related macrophage-pancreatic cancer cell interaction dialogue to promote the growth and metastasis of pancreatic cancer cells is clarified by adopting liposome carrier transfection, a pancreatic cancer cell and macrophage co-culture system and other means.
In the verification process, in-vitro and in-vivo experiments for promoting the growth and metastasis of pancreatic cancer cells by Cyr 61-regulated tumor-associated macrophage-pancreatic cancer cell interactive dialogue are needed. The experiment specifically comprises the steps of pancreatic cancer specimen collection and gene expression analysis, cyr61 eukaryotic expression plasmid construction, cyr61 stable transfection cell line establishment, cyr61 conditioned medium collection and the like, and the specific steps of Cyr61 stable transfection cell line establishment and Cyr61 conditioned medium collection are as follows: the pcDNA3.1-Cyr61 plasmid is transfected into pancreatic cancer cells (PANC-1 and BxPC-3), after 24 hours of transfection, the cells are transferred into a culture medium containing G418 antibiotics, and are cultured for 2 to 3 weeks to obtain pancreatic cancer cell clones resisting G418, and the cell clones are digested and mixed with enzyme to obtain a Cyr61 stable transfected cell line; the transfected cells are cultured in a serum-free culture medium for 24 hours, then the culture solution is collected, the centrifugation is performed for 5 minutes at 1000 revolutions, cell fragments are removed, and the supernatant is Cyr61 conditioned medium, and the cells are sub-packaged and frozen for preservation.
Centrifugation is used as an important ring in the Cyr61 conditioned medium, plays an important role in subsequent experiments, however, in the use process of the conventional centrifugal device, the supernatant is inconvenient to discharge, and after centrifugation is finished, the supernatant is lack of further filtration, so that the quality of the supernatant is poor, and the subsequent experiments are influenced.
Disclosure of Invention
First, the technical problem to be solved
The invention can solve the problems that the existing centrifugal device is inconvenient in discharging supernatant and the quality of the supernatant is poor due to the lack of further filtering the supernatant after centrifugation is finished, thereby influencing the subsequent experiment.
(II) technical scheme
In order to achieve the above purpose, the present invention adopts the following technical scheme, and in a first aspect, the present invention provides a centrifugal device for collecting Cyr61 target conditioned medium, which comprises a support assembly, a centrifugal assembly, a filtering assembly and a liquid taking assembly.
The support assembly comprises a base, a shell and a support frame, wherein the shell is installed on the upper portion of the base, the interior of the shell is hollow, and the support frame is installed on the upper portion of the shell.
The centrifugal assembly comprises a driving motor, a connecting shaft, a supporting arm and a fixing piece, wherein the driving motor is installed on the upper portion of the base, the connecting shaft is installed at the output end of the driving motor, the connecting shaft stretches into the shell, the supporting arm is installed on the side portion of the connecting shaft, and the fixing piece is installed on the upper portion of the supporting arm.
The filter component comprises an electric push rod, a connecting frame and a plurality of filter pieces, wherein the electric push rod is installed on the upper center portion of the supporting frame, the connecting frame is installed at the output end of the electric push rod, the filter pieces are respectively installed on the lower portions of the connecting frame, and the lower portions of the filter pieces are opposite to the upper portions of the fixing pieces.
The liquid taking component comprises a suction pump, a main pipe, a plurality of branch pipes and a plurality of suction heads, wherein the suction pump is arranged on the upper portion of the supporting frame, the main pipe is connected to the liquid inlet end of the suction pump, the branch pipes are respectively connected to the liquid inlet end of the main pipe, the suction heads are respectively arranged on the liquid inlet ends of the branch pipes, the branch pipes and the suction heads are respectively arranged on the side portions of the connecting frame, and the suction heads are respectively adjacent to the filtering pieces.
As a preferable technical scheme of the invention, the middle part of the base is hollow, and the outer contour of the whole section of the base is isosceles trapezoid.
As a preferable technical scheme of the invention, a through hole matched with the connecting shaft is formed in the middle of the shell, and the inner wall of the shell is in clearance fit with the side part of the supporting arm.
As a preferable technical scheme of the invention, the fixing piece comprises a supporting disc and a plurality of limiting rings, wherein the supporting disc is arranged on the upper part of the supporting arm, the upper part of the supporting disc is provided with a plurality of grooves, and the limiting rings are respectively arranged on the inner sides of the supporting disc adjacent to the grooves.
As a preferable technical scheme of the invention, the center of the supporting disc and the center of the connecting shaft are coaxially arranged, and the lower parts of the grooves are respectively opposite to the lower parts of the filtering pieces.
As a preferable technical scheme of the invention, the filter element comprises a connecting sleeve, a primary filter screen and a high-efficiency filter screen, wherein the connecting sleeve is arranged at the lower part of the connecting frame, the primary filter screen and the high-efficiency filter screen are respectively arranged at the inner side of the connecting sleeve, and the high-efficiency filter screen is arranged at the upper part of the primary filter screen.
As a preferable technical scheme of the invention, a sealing ring is glued on the outer side of the connecting sleeve, a guide part is arranged at the lower part of the sealing ring, and the section of the guide part is arranged in a conical shape.
As a preferable embodiment of the present invention, the main pipe is provided as a hose.
As a preferable technical scheme of the invention, a pipe clamp matched with the branch pipe is arranged at the side part of the connecting frame, and a flexible pad is arranged at the inner side of the pipe clamp.
In addition, the invention also provides a centrifugation method of the centrifugation device for collecting the Cyr61 target spot conditioned medium, which comprises the following steps:
s1, fixing a test tube: placing culture solution to be centrifuged into a plurality of test tubes, wherein the height of the culture solution is not more than two thirds of the total height of the test tubes, and then fixing the test tubes on the upper part of the fixing piece;
s2, centrifuging a culture solution: the driving motor is operated, the driving motor drives the connecting shaft, the supporting arm and the fixing piece to rotate, so that the test tubes rotate, broken cells are gathered on the side walls of the test tubes under the action of centrifugal force, and then the test tubes are kept stand for 1-3 h, so that the broken cells are gathered at the bottoms of the test tubes under the action of gravity;
s3, filtering the culture solution: operating an electric push rod, wherein the electric push rod drives the connecting frame to move downwards, so that a plurality of filtering pieces are inserted into a plurality of test tubes, and filtering culture solution again in the descending process of the filtering pieces, and filtering non-fallen broken cells to the bottoms of the test tubes;
s4, extracting supernatant, wherein in the process of downward movement of the connecting frame, the suction head stretches into the test tube, then the suction pump is operated, the operation of the suction pump sucks the supernatant on the upper portion of the test tube, and the supernatant sequentially passes through the suction head, the branch pipe, the main pipe and the suction pump and is discharged into an external storage container.
(III) beneficial effects
1. The invention provides a centrifugal device for collecting Cyr61 target spot condition culture medium, wherein a supporting component comprises a base, a shell and a supporting frame, the supporting component is prevented from being arranged on a laboratory table to be used through the base, the whole supporting is convenient, the shell provides a centrifugal space, and the supporting frame is convenient for supporting a filtering component and a liquid taking component;
2. the centrifugal device for collecting the Cyr61 target point condition culture medium comprises a centrifugal component, a centrifugal device and a centrifugal device, wherein the centrifugal component comprises a driving motor, a connecting shaft, a supporting arm and a fixing piece, centrifugal power is provided by the driving motor, a test tube is fixed by the fixing piece, the fixing piece is effectively supported by the cooperation of the connecting shaft and the supporting arm, and the stability of a centrifugal process is ensured;
3. the invention provides a centrifugal device for collecting Cyr61 target spot condition culture medium, wherein a filter assembly comprises an electric push rod, a connecting frame and a plurality of filter pieces, and the electric push rod operates to drive the connecting frame to lift, so that the filter pieces lift, and the filter pieces can conveniently extend into a test tube;
the invention provides a centrifugal device for collecting Cyr61 target point condition culture medium, wherein a liquid taking component comprises a suction pump, a main pipe, a plurality of branch pipes and a plurality of suction heads, the suction heads extend into a test tube in the downward moving process of a connecting frame, then the suction pump is operated, the operation of the suction pump sucks supernatant liquid on the upper part of the test tube, and the supernatant liquid sequentially passes through the suction heads, the branch pipes, the main pipe and the suction pump and is discharged into an external storage container.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some examples of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic view in front cross-section of the present invention;
FIG. 2 is a schematic front view of a portion of the structure of the support plate of the present invention;
FIG. 3 is a schematic top view of a portion of the structure of the support disk of the present invention;
FIG. 4 is an enlarged schematic view of the invention at A;
FIG. 5 is a schematic view of a pipe clamp portion of the present invention.
In the figure: 100. a support assembly; 110. a base; 120. a housing; 121. a through hole; 130. a support frame; 200. a centrifuge assembly; 210. a driving motor; 220. a connecting shaft; 230. a support arm; 240. a fixing member; 241. a support plate; 242. a limiting ring; 243. a groove; 300. a filter assembly; 310. an electric push rod; 320. a connecting frame; 321. a pipe clamp; 322. a flexible pad; 330. a filter; 331. connecting sleeves; 332. a primary filter screen; 333. an efficient filter screen; 334. a seal ring; 335. a guide part; 400. a liquid taking component; 410. a suction pump; 420. a main pipe; 430. a branch pipe; 440. suction head.
The achievement of the objects, functional features and advantages of the present invention will be further described with reference to the accompanying drawings, in conjunction with the embodiments.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, based on the embodiments of the invention, which are apparent to those of ordinary skill in the art without inventive faculty, are intended to be within the scope of the invention.
Thus, the following detailed description of the embodiments of the invention, as presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, based on the embodiments of the invention, which are apparent to those of ordinary skill in the art without inventive faculty, are intended to be within the scope of the invention.
It should be noted that: like reference numerals and letters denote like items in the following figures, and thus once an item is defined in one figure, no further definition or explanation thereof is necessary in the following figures.
In the description of the present invention, it should be understood that the terms "longitudinal," "upper," "lower," "left," "right," "top," "bottom," "inner," "outer," and the like indicate orientations or positional relationships based on the orientation or positional relationships shown in the drawings, merely to facilitate description of the present invention and simplify the description, and do not indicate or imply that the devices or elements being referred to must have a particular orientation, be configured and operated in a particular orientation, and thus should not be construed as limiting the present invention.
Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include one or more such feature. In the description of the present invention, the meaning of "a plurality" is two or more, unless explicitly defined otherwise.
Example 1
As shown in fig. 1 to 5, a centrifuge for collecting Cyr61 target conditioned medium includes a support assembly 100, a centrifuge assembly 200, a filter assembly 300, and a liquid extraction assembly 400.
The support assembly 100 comprises a base 110, a shell 120 and a support frame 130, wherein the shell 120 is arranged at the upper part of the base 110, and when the support assembly is specifically arranged, in order to achieve a better overall support effect, the middle part of the base 110 is hollow, and the outline of the overall section of the base 110 is isosceles trapezoid; the inside of the housing 120 is hollow, the upper portion of the housing 120 is open, and the supporting frame 130 is mounted on the upper portion of the housing 120 by bolts.
The centrifugal assembly 200 includes a driving motor 210, a connection shaft 220, a support arm 230, and a fixing member 240, the driving motor 210 is installed at the upper portion of the base 110, the connection shaft 220 is installed at the output end of the driving motor 210, the connection shaft 220 extends into the housing 120, the support arm 230 is installed at the side portion of the connection shaft 220, and the fixing member 240 is installed at the upper portion of the support arm 230.
When the support arm 230 is specifically configured, in order to enable the connecting shaft 220 to effectively rotate, the middle portion of the housing 120 is provided with a through hole 121 matched with the connecting shaft 220, and in order to ensure that the support arm 230 effectively rotates, the inner wall of the housing 120 is in clearance fit with the side portion of the support arm 230.
In this embodiment, the fixing member 240 includes a support plate 241 and a plurality of limiting rings 242, the support plate 241 is mounted on the upper portion of the support arm 230, a plurality of grooves 243 are formed on the upper portion of the support plate 241, the grooves 243 are arranged in an annular array relative to the center of the support plate 241, the plurality of limiting rings 242 are respectively mounted on the inner sides of the support plate 241 adjacent to the grooves 243, and the limiting rings 242 are elastic rubber rings, so that the fixing of test tubes can be better realized; when the test tube fixing device is specifically arranged, the center of the supporting plate 241 and the center of the connecting shaft 220 are coaxially arranged, and when the test tube fixing device is specifically used, a plurality of test tubes are respectively inserted into the grooves 243 and are limited and fixed through the limiting rings 242, so that good fixing of the test tubes is achieved.
The filter assembly 300 includes an electric push rod 310, a connecting frame 320 and a plurality of filter elements 330, the electric push rod 310 is mounted on the upper center portion of the supporting frame 130 through bolts, the connecting frame 320 is mounted on the output end of the electric push rod 310, the plurality of filter elements 330 are respectively mounted on the lower portions of the connecting frame 320, the lower portions of the plurality of filter elements 330 are opposite to the upper portions of the fixing elements 240, and when the filter assembly is specifically arranged, the lower portions of the plurality of grooves 243 are opposite to the lower portions of the plurality of filter elements 330.
In this embodiment, the filter 330 includes a connecting sleeve 331, a primary filter screen 332 and a high-efficiency filter screen 333, the connecting sleeve 331 is mounted at the lower part of the connecting frame 320, the primary filter screen 332 and the high-efficiency filter screen 333 are respectively mounted at the inner side of the connecting sleeve 331, and the high-efficiency filter screen 333 is positioned at the upper part of the primary filter screen 332; when the device is specifically arranged, the sealing ring 334 is glued on the outer side of the connecting sleeve 331, the guide part 335 is arranged at the lower part of the sealing ring 334, the section of the guide part 335 is in a conical shape, the guide part 335 guides, the sealing ring 334 is more convenient to insert into a test tube, and the guide part 335 and the sealing ring 334 are integrally arranged; when the culture medium filter is specifically used, the connecting sleeve is lifted along with the lifting of the connecting frame 320, is inserted into the test tube, is in contact with the inner wall of the test tube by utilizing the sealing ring 334 to seal, and is matched with the primary filter screen 332 and the efficient filter screen 333 to filter culture medium.
The liquid taking assembly 400 comprises a suction pump 410, a main pipe 420, a plurality of branch pipes 430 and a plurality of suction heads 440, wherein the suction pump 410 is arranged at the upper part of the support frame 130, the main pipe 420 is connected with the liquid inlet end of the suction pump 410, the main pipe 420 is provided as a hose, the plurality of branch pipes 430 are respectively connected with the liquid inlet end of the main pipe 420, the plurality of suction heads 440 are respectively arranged at the liquid inlet ends of the plurality of branch pipes 430, the plurality of branch pipes 430 and the plurality of suction heads 440 are respectively arranged at the side parts of the connecting frame 320, when the liquid taking assembly is specifically arranged, the side parts of the connecting frame 320 are provided with pipe clamps 321 matched with the branch pipes 430, and the inner sides of the pipe clamps 321 are provided with flexible pads 322; a plurality of suction heads 440 are respectively adjacent to the plurality of filter elements 330.
In addition, the invention also provides a centrifugation method of the centrifugation device for collecting the Cyr61 target spot conditioned medium, which comprises the following steps:
s1, fixing a test tube: placing culture solution to be centrifuged into a plurality of test tubes, wherein the height of the culture solution is not more than two thirds of the total height of the test tubes, and then fixing the plurality of test tubes on the upper part of the fixing piece 240;
s2, centrifuging a culture solution: the driving motor 210 is operated, the driving motor 210 is operated to drive the connecting shaft 220, the supporting arm 230 and the fixing piece 240 to rotate, so that a plurality of test tubes rotate, broken cells are gathered on the side walls of a plurality of test tubes under the action of centrifugal force, and then the test tubes are kept stand for 1-3 h, so that the broken cells are gathered at the bottoms of the test tubes under the action of gravity;
s3, filtering the culture solution: operating the electric push rod 310, wherein the electric push rod 310 drives the connecting frame 320 to move downwards, so that the plurality of filtering pieces 330 are inserted into the plurality of test tubes, and filtering the culture solution again in the descending process of the filtering pieces 330, and filtering the broken cells which do not fall to the bottom of the test tubes;
s4, extracting supernatant, wherein in the process of downward movement of the connecting frame 320, the suction head 440 stretches into the test tube, then the suction pump 410 is operated, the operation of the suction pump 410 sucks the supernatant on the upper part of the test tube, and the supernatant sequentially passes through the suction head 440, the branch pipe 430, the main pipe 420 and the suction pump 410 and is discharged into an external storage container.
In summary, the invention simply and conveniently realizes the separation of the culture solution, and simultaneously, after the separation is finished, the secondary filtration and the suction of the supernatant liquid are carried out, thereby achieving good separation effect and ensuring the quality of the supernatant liquid.
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. A centrifuge device for collecting Cyr61 target conditioned medium, comprising:
the support assembly (100), the support assembly (100) comprises a base (110), a shell (120) and a support frame (130), the shell (120) is installed on the upper portion of the base (110), the interior of the shell (120) is hollow, and the support frame (130) is installed on the upper portion of the shell (120);
the centrifugal assembly (200), the centrifugal assembly (200) comprises a driving motor (210), a connecting shaft (220), a supporting arm (230) and a fixing piece (240), the driving motor (210) is installed on the upper portion of the base (110), the connecting shaft (220) is installed at the output end of the driving motor (210), the connecting shaft (220) stretches into the shell (120), the supporting arm (230) is installed on the side portion of the connecting shaft (220), and the fixing piece (240) is installed on the upper portion of the supporting arm (230);
the filtering assembly (300) comprises an electric push rod (310), a connecting frame (320) and a plurality of filtering pieces (330), wherein the electric push rod (310) is installed at the central upper part of the supporting frame (130), the connecting frame (320) is installed at the output end of the electric push rod (310), the filtering pieces (330) are respectively installed at the lower parts of the connecting frame (320), and the lower parts of the filtering pieces (330) are opposite to the upper parts of the fixing pieces (240);
the liquid taking assembly (400), the liquid taking assembly (400) comprises a suction pump (410), a main pipe (420), a plurality of branch pipes (430) and a plurality of suction heads (440), wherein the suction pump (410) is installed on the upper portion of the supporting frame (130), the main pipe (420) is connected to the liquid inlet end of the suction pump (410), the plurality of branch pipes (430) are respectively connected to the liquid inlet end of the main pipe (420), the plurality of suction heads (440) are respectively installed on the liquid inlet ends of the plurality of branch pipes (430), the plurality of branch pipes (430) and the plurality of suction heads (440) are respectively installed on the side portions of the connecting frame (320), and the plurality of suction heads (440) are respectively adjacent to the plurality of filtering pieces (330);
the fixing piece (240) comprises a supporting disc (241) and a plurality of limiting rings (242), the supporting disc (241) is arranged on the upper portion of the supporting arm (230), a plurality of grooves (243) are formed in the upper portion of the supporting disc (241), and the limiting rings (242) are respectively arranged on the inner sides, adjacent to the grooves (243), of the supporting disc (241);
the filter element (330) comprises a connecting sleeve (331), a primary filter screen (332) and a high-efficiency filter screen (333), wherein the connecting sleeve (331) is arranged at the lower part of the connecting frame (320), the primary filter screen (332) and the high-efficiency filter screen (333) are respectively arranged at the inner side of the connecting sleeve (331), and the high-efficiency filter screen (333) is arranged at the upper part of the primary filter screen (332).
2. The centrifuge device for collecting Cyr61 target conditioned media of claim 1, wherein: the middle part of base (110) is the cavity setting, the outline of the whole section of base (110) is isosceles trapezoid setting.
3. The centrifuge device for collecting Cyr61 target conditioned media of claim 1, wherein: the middle part of the shell (120) is provided with a through hole (121) matched with the connecting shaft (220), and the inner wall of the shell (120) is in clearance fit with the side part of the supporting arm (230).
4. The centrifuge device for collecting Cyr61 target conditioned media of claim 1, wherein: the center of the supporting disc (241) is coaxially arranged with the center of the connecting shaft (220), and the lower parts of the grooves (243) are respectively opposite to the lower parts of the filtering pieces (330).
5. The centrifuge device for collecting Cyr61 target conditioned media of claim 1, wherein: the outside of adapter sleeve (331) glues has sealing ring (334), the lower part of sealing ring (334) is provided with guiding portion (335), the section of guiding portion (335) is the taper setting.
6. The centrifuge device for collecting Cyr61 target conditioned media of claim 1, wherein: the main pipe (420) is provided as a hose.
7. The centrifugal device for collecting Cyr61 target conditioned media according to claim 1, wherein a pipe clamp (321) matched with the branch pipe (430) is arranged at the side part of the connecting frame (320), and a flexible pad (322) is arranged at the inner side of the pipe clamp (321).
8. A centrifugation method using the centrifugation device for collecting Cyr61 target conditioned medium according to any one of claims 1 to 7, comprising the steps of:
s1, fixing a test tube: placing culture solution to be centrifuged into a plurality of test tubes, wherein the height of the culture solution is not more than two thirds of the total height of the test tubes, and then fixing the test tubes on the upper part of the fixing piece;
s2, centrifuging a culture solution: the driving motor is operated, the driving motor drives the connecting shaft, the supporting arm and the fixing piece to rotate, so that the test tubes rotate, broken cells are gathered on the side walls of the test tubes under the action of centrifugal force, and then the test tubes are kept stand for 1-3 h, so that the broken cells are gathered at the bottoms of the test tubes under the action of gravity;
s3, filtering the culture solution: operating an electric push rod, wherein the electric push rod drives the connecting frame to move downwards, so that a plurality of filtering pieces are inserted into a plurality of test tubes, and filtering culture solution again in the descending process of the filtering pieces, and filtering non-fallen broken cells to the bottoms of the test tubes;
s4, extracting supernatant, wherein in the process of downward movement of the connecting frame, the suction head stretches into the test tube, then the suction pump is operated, the operation of the suction pump sucks the supernatant on the upper portion of the test tube, and the supernatant sequentially passes through the suction head, the branch pipe, the main pipe and the suction pump and is discharged into an external storage container.
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| WO2020138506A1 (en) * | 2018-12-27 | 2020-07-02 | エイブル株式会社 | Perfusion culture apparatus and centrifugal separator |
| CN210543707U (en) * | 2019-05-20 | 2020-05-19 | 湖北锦顺祥环保有限公司 | Reverse suction device of fiber rotary disc filter |
| CN111304056A (en) * | 2020-03-11 | 2020-06-19 | 李容尔 | Cell sample draws partial shipment device |
| CN112034194A (en) * | 2020-09-14 | 2020-12-04 | 安徽新生医学科技有限公司 | Automatic sample feeding device for measuring selenium content of hair based on fluorescence spectrum |
| CN113214970A (en) * | 2021-05-07 | 2021-08-06 | 锦州医科大学 | Self-adaptive adjustment somatic cell culture device and culture method thereof |
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