CN110804548B - Cell co-culture equipment - Google Patents
Cell co-culture equipment Download PDFInfo
- Publication number
- CN110804548B CN110804548B CN201911141962.8A CN201911141962A CN110804548B CN 110804548 B CN110804548 B CN 110804548B CN 201911141962 A CN201911141962 A CN 201911141962A CN 110804548 B CN110804548 B CN 110804548B
- Authority
- CN
- China
- Prior art keywords
- culture dish
- type
- type culture
- cell
- dish body
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- 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
- C12M23/10—Petri dish
-
- 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
- C12M23/34—Internal compartments or partitions
-
- 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
- C12M23/48—Holding appliances; Racks; Supports
-
- 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
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/02—Membranes; Filters
-
- 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
- C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
- C12M37/04—Seals
-
- 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
-
- 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/34—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of gas
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Sustainable Development (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Clinical Laboratory Science (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention belongs to the technical field of biological medicine, and relates to cell culture equipment, in particular to cell co-culture equipment. The equipment comprises a B-type culture dish, an A-type culture dish and a lifting unit, wherein a microporous membrane is arranged on the A-type culture dish, the lifting unit controls the A-type culture dish to move in the vertical direction and can control the communication time between the B-type culture dish and the A-type culture dish, so that the control of the time of information exchange among different cells is realized. The scheme can be applied to experimental study of cell co-culture, and can be used for achieving research of influence of various cell co-culture and cell spacing on cell information exchange, direct cell contact co-culture and the like.
Description
Technical Field
The invention belongs to the technical field of biological medicine, and relates to cell culture equipment, in particular to cell co-culture equipment.
Background
The cell co-culture technology is to co-culture two or more than two kinds of cells in the same environment, so that the cells can communicate information with each other and support each other to grow and proliferate. The cell co-culture technology cultures different cells in the same system, simulates the microenvironment of cell growth, and provides an effective research means for researching cell-cell interaction. A large number of cell co-culture techniques and apparatus have emerged in the prior art, with transwell techniques being the most widely used. Conventional Transwell equipment includes a cell with a polycarbonate microporous membrane and a petri dish with a culture tank. In the experiment, the cell is placed in a culture tank of a culture dish, two kinds of cells are respectively inoculated in the cell and the culture tank, and as the two kinds of cells are separated by a polycarbonate membrane, the polycarbonate membrane can pass small molecules, the cells can not pass through, and the culture medium components in the cell and the culture medium components in the culture tank can respectively influence the cells in the culture tank and the cells in the cell, so that the influence of cell secretion substances on the growth, the morphology and the gene or protein expression of the other kind of cells can be studied.
However, the existing transwell technology has the following disadvantages: the two cell growth environments are not independent of each other, and the material communication between the two cells cannot be controlled; the distance between two cells is not adjustable, and the influence of cell spacing on cell information communication cannot be studied; only indirect co-culture (transfer of information substances through the medium) can be performed, and direct co-culture of two adherent cells (direct contact between two cells can occur) cannot be achieved; co-culture of two or more cells cannot be performed.
Disclosure of Invention
The invention aims to provide a cell co-culture device which can control the time of information communication among different cells.
In order to solve the technical problems, the technical scheme of the invention is as follows:
a cell co-culture device comprises a type B culture dish, a type A culture dish and a lifting unit; the A-type culture dish comprises an A-type culture dish body, a sealing disc, a hanging lug and a vertical rod, wherein a microporous layer is fixed in the A-type culture dish body, and the periphery of the microporous layer is fixedly connected with the inner wall of the A-type culture dish body in a sealing manner; the hanging lugs are fixed on the A-type culture dish body, the number of the hanging lugs is a plurality, and the hanging lugs are in axisymmetric distribution by taking the central axis of the A-type culture dish body as a symmetry axis; the sealing disc is used for sealing the lower end of the A-type culture dish body, the sealing disc is detachable from the A-type culture dish body, the sealing disc is fixedly connected with the vertical rod, and the vertical rod is fixed on the lifting unit; the B-type culture dish comprises a culture tank which is positioned right below the A-type culture dish body, and the hanging lugs are used for hanging the A-type culture dish body in the culture tank; the lifting unit is used for driving the vertical rod to move in the vertical direction and driving the sealing disc to fall off from the A-type culture dish body.
By adopting the technical scheme, the technical principle is as follows: in performing cell co-culture experiments, a cells were seeded in a type a dish body and B cells were seeded in a type B dish (two cells are designated as a cells and B cells). The cells A and B are adherent cells, and under the action of gravity, the cells A and B can be attached to the bottom of the culture dish for growth. The lifting unit is adjusted to enable the A-type culture dish body to descend, the culture tank is located under the A-type culture dish body, the A-type culture dish body moves downwards, the whole A-type culture dish body can be inserted into the culture tank until the hanging lugs are in contact with the upper surface of the B-type culture dish, and the lifting unit is stopped. When the A cells and the B cells grow to a certain degree and the two cells are required to communicate information. The lifting unit is adjusted to descend, the hanging lugs hang the A-type culture dish body above the B-type culture dish, upward acting force is applied to the A-type culture dish body, the lifting unit applies downward acting force to the sealing disc through the connecting rod, the A-type culture dish body is separated from the sealing disc, and A cells and B cells can communicate substances through the microporous layer.
The beneficial effects are that:
(1) By adopting the scheme, the time of information communication between two cells can be controlled. When the cell culture device is suitable, the lifting mechanism is adjusted downwards, so that the sealing disc is separated from the A cell culture body, and the A cells and the B cells start to communicate information through the microporous layer (the information carrier is various signal factors secreted by the cells). Under the drive of elevating system, make sealing disk drop from the A cell culture body, make both separate more labour saving and time saving for manual.
(2) In the scheme, when cells are cultured, the A-type culture dish is placed inside the B-type culture dish, and compared with the scheme that two cells are separately cultured and put together at proper time to perform co-culture, the scheme creates the same growth environment (comprising temperature, humidity and the like) for the two cells, avoids the difference of growth conditions of the two cells caused by the difference of culture conditions, and therefore increases the accuracy of experiments. In addition, the volume of the A-type culture dish is smaller than that of the B-type culture dish, and the A-type culture dish is easy to be interfered by external environment, but the A-type culture dish is placed in the B-type culture dish, so that the interference can be reduced, and the experimental accuracy is improved.
(3) In this scheme, A type culture dish is located the type B culture dish directly over, has the one-to-one correspondence, has avoided mixing up the culture dish, puts the condition in the type B culture dish with the type A culture dish that does not correspond.
(4) The A-type culture dish body is automatically put in and taken out through the mechanical structure, so that the manual putting in and taking out of the A-type culture dish body by forceps is avoided, and the operation difficulty and the labor intensity are reduced.
Further, the microporous layer is a polycarbonate film.
By adopting the technical scheme, the polycarbonate membrane is a conventional cell co-culture membrane material, is easy to obtain, and is safe and nontoxic.
Further, the microporous layer has a microporous diameter of 0.5 to 8 μm.
By adopting the technical scheme, the cultured cells can be ensured not to pass through the microporous layer, but small molecular substances such as various growth factors secreted by the cells can pass through the microporous layer, so that the interaction among the cells is realized.
Further, the number of the culture tanks of the type B culture dishes is a plurality, and the number of the type A culture dishes is consistent with the number of the culture tanks.
By adopting the technical scheme, a plurality of culture tanks and A-type culture dishes can be arranged for simultaneously carrying out cell co-culture of a plurality of groups, so that the experimental efficiency is improved.
Further, the culture tank is a cylindrical tank with an opening at the upper part, and the A-shaped culture dish body is cylindrical.
By adopting the technical scheme, compared with a square structure, the cylindrical structure is more suitable for cell culture, and the cells are easily gathered at corners by using the square culture dish, so that the difficulty is caused to harvest or passage cells.
Further, the outside of A type culture dish body lateral wall is equipped with two sets of hangers fixed slot groups that are used for inserting the hangers, and the hangers fixed slot group includes a plurality of hangers fixed slot, the axial distribution of A type culture dish body is followed to the hangers fixed slot, and two sets of hangers fixed slot are axisymmetric distribution with the axis of A type culture dish body as the symmetry axis.
By adopting the technical scheme, the depth of the A-shaped culture dish body placed in the culture tank can be adjusted. Because be equipped with a plurality of hangers fixed slots on the A type culture dish body, can insert the hangers in the hangers fixed slot of different positions to adjust the degree of depth that the A type culture dish body inserted the culture tank. The deeper the A-type culture dish body is inserted into the culture tank, the closer the distance between two cells is, and the arrangement can be used for researching the influence of the distance between the cells on the cell interaction.
The hanger fixed slot takes the middle shaft of the A-type culture dish body as the shaft to be axisymmetrically distributed, so that the balance of the A-type culture dish body is conveniently maintained, and the shaking and the inclination of the A-type culture dish body are reduced when the A-type culture dish body is hung in the culture slot.
Further, the lifting unit comprises a lifting beam, a screw rod mechanism and a base; the vertical rod is detachably fixed on the lifting beam, the base is used for fixing a screw rod mechanism and placing a B-type culture dish, and the screw rod mechanism is used for lifting the lifting beam; the lifting beam is horizontally arranged and is rotationally connected with the screw rod mechanism.
By adopting the technical scheme, when adding liquid or inoculating into the A-type culture dish, the lifting beam is rotated, so that the A-type culture dish is not positioned right above the B-type culture dish, and reagents, cells or culture mediums are prevented from being carelessly dripped into the B-type culture dish, and the occurrence of the condition of polluting the B-type culture dish is avoided.
Further, a placing groove for placing the B-type culture dish is formed in the base.
By adopting the technical scheme, the B-type culture dish can be stably placed, and the B-type culture dish is prevented from toppling over.
Further, a partition board is arranged in the A-type culture dish body and is perpendicular to the microporous layer, and the partition board is used for dividing the accommodating cavity of the A-type culture dish body into a plurality of cavities.
By adopting the technical scheme, a plurality of cells can be simultaneously cultured in the A-type culture dish body, and the interaction among the plurality of cells is studied (see the embodiment 3 for details).
Further, the microporous layer is positioned in the middle of the A-type culture dish body
By adopting the technical scheme, the mutual influence of two cells under the condition of no contact can be tested, and the mutual influence of two cells under the condition of contact can also be tested (see the example 2 for details).
Drawings
FIG. 1 is a front view of the cell co-culture apparatus of example 1.
FIG. 2 is a longitudinal sectional view of the type B dish of example 1 (the state in which the type A dish is not placed).
FIG. 3 is a longitudinal cross-sectional view of a type B dish in which a type A dish was placed in example 1.
FIG. 4 is a top view of the dish cover of example 1.
Fig. 5 is a top view of the slide connection sheet of embodiment 1.
FIG. 6 is a longitudinal cross-sectional view of the type A dish of example 2.
FIG. 7 is a top view of the body of the type A dish of example 3.
Detailed Description
The following is a further detailed description of the embodiments:
reference numerals in the drawings of the specification include: the culture dish comprises a base 1, a type B culture dish 2, a type A culture dish body 3, a sealing disc 4, a connecting rod 5, hanging lugs 6, a fixing rod 7, a lifting beam 8, a screw rod 9, a vertical supporting rod 10, an upper transverse plate 11, a lower transverse plate 12, a gear 13, a worm 14, a culture tank 15, a hanging lug fixing groove 16, a culture dish cover 17, a microporous layer 18, a sliding block 19, a sliding connecting piece 20, a lifting beam connecting piece 21, a fixing rod nut 22, a fixing rod limiting piece 23, a handle 24, a placing groove 25, a screw rod limiting piece 26, a screw rod nut 27 and a partition plate 28.
Example 1:
as shown in FIG. 1, a cell co-culture apparatus includes a type B dish 2, a type A dish, and a lifting unit. As shown in FIG. 2, four culture tanks 15 are arranged in one B-type culture dish 2, the four culture tanks 15 are positioned on the same straight line, and the culture tanks 15 are cylindrical tanks. As shown in fig. 1, the a-type culture dish comprises an a-type culture dish body 3, a sealing disk 4, a hanging lug 6 and a vertical rod. As shown in FIG. 3, the A-type culture dish body 3 is cylindrical, the upper part is opened, the lower part is fixedly provided with a microporous layer 18, and the periphery of the microporous layer 18 is fixedly connected with the inner wall of the A-type culture dish body 3 in a sealing way. The microporous layer 18 is a polycarbonate membrane, and is a membrane material used in conventional transwell experiments, the diameter of micropores of the microporous layer 18 can be between 0.5 μm and 8 μm, and in this embodiment, the diameter of micropores in the microporous layer 18 is 3 μm, so that cultured cells can not pass through the microporous layer 18, but small molecular substances such as various growth factors secreted by the cells can pass through the microporous layer 18, so as to realize interaction among cells. The lower extreme of A type culture dish body 3 is fixed with sealing disk 4 through the block, and sealing disk 4 seals the lower extreme of A type culture dish body 3, and under the exogenic action, A type culture dish body 3 and sealing disk 4 separable. A vertical rod is fixed on the sealing disc 4 and is divided into a connecting rod 5 and a fixing rod 7, and one end of the connecting rod 5 and the sealing disc 4 are integrally formed. The other end of the connecting rod 5 is fixed with the fixing rod limiting piece 23, the fixing rod 7 is also fixed with the fixing rod limiting piece 23, and the connecting rod 5, the fixing rod limiting piece 23 and the fixing rod 7 are integrally formed. Two sets of hanging lug fixing groove sets for inserting hanging lugs 6 are arranged on the outer side of the side wall of the A-type culture dish body 3, and each set of hanging lug fixing groove sets comprises a plurality of hanging lug fixing grooves 16 (5 hanging lug fixing grooves in the embodiment). The hanging lug fixing grooves 16 are distributed along the axial direction of the A-shaped culture dish body 3, and the two hanging lug fixing groove groups are symmetrically distributed by taking the central axis of the A-shaped culture dish body 3 as an axis. The hanger fixing groove 16 is a square groove, the hanger 6 is of a sheet structure with a thickness, two hangers 6 are respectively inserted into the corresponding hanger fixing groove 16, the distance between the non-insertion ends of the two hangers 6 is larger than the diameter of the culture groove 15 of the B-type culture dish 2, and the A-type culture dish body 3 can be hung in the culture groove 15 of the B-type culture dish 2. The hanging lug fixing grooves 16 are distributed along the axial direction of the A-shaped culture dish body 3, so that the depth of the A-shaped culture dish body 3 placed in the culture groove 15 can be adjusted; the hanging lug fixing grooves 16 are symmetrically distributed by taking the central shaft of the A-shaped culture dish body 3 as an axis, and the two hanging lugs 6 are positioned on the same plane, so that the balance of the A-shaped culture dish body 3 is conveniently maintained, and shaking and inclination of the A-shaped culture dish body 3 are reduced when the A-shaped culture dish body 3 is hung in the culture groove 15.
As shown in fig. 1, the lifting unit includes a lifting beam 8, a screw mechanism, and a base 1. The base 1 is provided with a placement groove 25 (shown in fig. 3) for placing the B-type culture dish 2. The screw mechanism comprises a screw 9, a slide block 19, an adjusting gear set and a supporting frame. The support frame comprises a vertical support rod 10, an upper transverse plate 11 and a lower transverse plate 12. The vertical support rod 10 is vertically arranged, the right end horizontal welding of the upper transverse plate 11 is arranged at the top end of the vertical support rod 10, the lower side of the upper transverse plate is welded with a supporting foot, and the lower surface of the supporting foot and the lower surface of the base 1 are arranged on the same horizontal plane. The lower part of the vertical supporting rod 10 and the upper part of the supporting leg are welded with a lower transverse plate 12, the lower transverse plate 12 is horizontally arranged, one end of the lower transverse plate 12 is welded with the vertical supporting rod 10, and the other end is welded with the base 1. The vertical support bar 10 is provided with a vertical sliding hole. The upper end of the screw rod 9 passes through the upper transverse plate 11 and is rotationally connected with the upper transverse plate 11; the lower end of the screw rod 9 passes through the lower transverse plate 12 and is rotationally connected with the lower transverse plate 12. The specific method is as follows: the upper and lower parts of the screw rod 9 are welded and fixed with screw rod limiting plates 26 (the screw rod 9 passes through the through holes on the screw rod limiting plates 26 and then is welded and fixed), and then the upper and lower parts of the screw rod 9 respectively pass through the through holes formed on the upper transverse plate 11 and the lower transverse plate 12 and are respectively connected with screw rod nuts 27 in a threaded manner. The diameter of the screw limiting piece 26 and the diameter of the screw nut 27 are both larger than the diameters of the through holes formed in the upper transverse plate 11 and the lower transverse plate 12, and after the screw nut 27 is mounted on the screw 9, the screw nut 27 and the screw limiting piece 26 limit the movement of the screw 9 in the vertical direction. The screw rod 9 is rotatably connected between the upper transverse plate 11 and the lower transverse plate 12, and the screw rod 9 can only rotate and cannot move in the vertical direction.
As shown in fig. 1, a threaded through hole is formed in the slider 19, and the screw 9 is screwed to the slider 19 through the threaded through hole in the slider 19. Two mutually parallel lifting beam connecting pieces 21 are fixed on the left side of the sliding block 19, the lifting beam 8 is rotatably connected between the two lifting beam connecting pieces 21 through a vertical rotating shaft, and the lifting beam 8 is horizontally arranged and rotates by taking the rotating shaft as an axis. The two ends of the rotating shaft respectively pass through the upper lifting beam connecting sheet 21 and the lower lifting beam connecting sheet 21 and are connected with the nuts in a threaded manner. The A-shaped culture dishes are fixed on the lifting beam 8 and are axially distributed along the lifting beam 8. The specific method is as follows: the fixing rod 7 passes through the through hole on the lifting beam 8 and is in threaded connection with the fixing rod nut 22, and the fixing rod nut 22 and the fixing rod limiting piece 23 fix the A-type culture dish on the lifting beam 8. The lifting beam 8 is rotated so that the A-type culture dish is positioned right above the culture tank 15 of the B-type culture dish 2. Two sliding connection pieces 20 are fixed on the right side of the sliding block 19, as shown in fig. 1 and 5, the left end of the sliding connection piece 20 is welded with the sliding block 19, a sliding shaft is welded between the two sliding connection pieces 20, and the sliding shaft is arranged in a sliding hole on the vertical supporting rod 10 in a penetrating manner and can slide in the vertical direction in the sliding hole.
The adjusting gear set is shown in fig. 1, and comprises a gear 13 and a worm 14, wherein the gear 13 is welded and fixed on the screw rod 9, and the gear 13 and the screw rod 9 are coaxially arranged. The worm 14 is welded and fixed on a central shaft, the worm 14 is coaxial with the central shaft, the worm 14 is horizontally arranged and vertical to the vertical supporting rod 10, and the worm 14 is meshed with the gear 13. Two ends of a central shaft coaxial with the worm 14 are respectively and rotatably connected to two support plates, the support plates are fixedly welded with the vertical support rods 10, and a handle 24 is fixedly welded at one end of the central shaft.
The specific embodiment is as follows:
when the cell co-culture experiment is carried out (two cells are denoted by A cells and B cells, the two cells are adherent cells, under the action of gravity, the A cells and the B cells are attached to the bottom of the culture dish for growth), the A-type culture dish is firstly fixed on the lifting beam 8 (the fixing rod 7 passes through a through hole on the lifting beam 8 and then is in threaded connection with the fixing rod nut 22, and the fixing rod nut 22 and the fixing rod limiting piece 23 fix the A-type culture dish on the lifting beam 8), the lifting beam 8 is rotated, so that the A-type culture dish is not positioned right above the B-type culture dish 2, and the reagent or the culture medium is prevented from being carelessly dripped into the B-type culture dish 2 when the A-type culture dish is added with liquid. Cells a were seeded into the type a dish body 3 and then B cells were seeded into the type B dish 2. The lifting beam 8 is rotated so that the a-type culture dish is located directly above the culture tank 15 of the B-type culture dish 2. Then, the worm 14 is rotated through the handle 24, the worm 14 drives the gear 13 to rotate, the lead screw 9 rotates to drive the sliding block 19 to move downwards, and the lifting beam 8 moves downwards to drive the A-type culture dish to move downwards. When the A-type culture dish descends to the position of the B-type culture dish 2, the hanging lugs 6 of the A-type culture dish contact the upper surface of the B-type culture dish 2, the rotation of the handle 24 is stopped, the culture dish cover 17 is covered (as shown in fig. 4, the diameter of the culture dish cover 17 is larger than that of the culture tank 15, and a notch is formed in the culture dish cover 17 for placing the connecting rod 5), and cell culture is carried out. Because the A-type culture dish body 3 is provided with the plurality of hanging lug fixing grooves 16, the hanging lugs 6 can be inserted into the hanging lug fixing grooves 16 at different positions so as to adjust the depth of the A-type culture dish body 3 inserted into the culture groove 15. But it is ensured that the two lugs 6 are located on the same horizontal plane so that the a-type culture dish will not tilt. The deeper the a-type culture dish body 3 is inserted into the culture bath 15, the closer the distance between a cells and B cells is, and this arrangement can be used to study the effect of the distance of the cells on the cell interaction. After a period of incubation (the incubation time is determined according to the specific cell type), both a cells and B cells have grown adherent. The A-type culture dish body 3 is placed in the B-type culture dish 2 to create the same growing environment (including temperature, humidity and the like) for two cells, so that the difference of growing conditions of the two cells caused by the difference of the culturing conditions is avoided, and the accuracy of the experiment is improved. In addition, the volume of the A-type culture dish is smaller than that of the B-type culture dish 2, and the A-type culture dish is easy to be interfered by external environment, but the A-type culture dish is placed in the B-type culture dish 2, so that the interference can be reduced, and the experimental accuracy is improved.
After a period of culture, both the A cells and the B cells are in a more vigorous growth state (exponential growth state), and an experiment of the interaction between cell secretions can be started. Before that, since the a cells and the B cells are in a non-vigorous growth state, if they are affected by each other's secretions, they have a certain negative effect on the growth of the cells, so that the sealing disk 4 seals the lower end of the a-type culture dish body 3, and the a cells and the B cells do not communicate with each other. When the cells are in an exponential growth state, the handle 24 is rotated to enable the A-type culture dish to continue to descend, at the moment, the hanging lugs 6 hang the A-type culture dish body 3 on the B-type culture dish 2, upward acting force is applied to the A-type culture dish body 3, downward acting force is applied to the sealing disc 4 by the lifting beam 8 through the connecting rod 5, the A-type culture dish body 3 and the sealing disc 4 are separated, and the A cells and the B cells can communicate substances through the microporous layer 18. After the cells are cultured for a certain period of time, the culture dish cover 17 is taken down, the handle 24 is rotated to enable the lifting beam 8 to ascend, and the sealing disc 4 supports the A-type culture dish body 3 upwards, so that the A-type culture dish body 3 is taken out. In this case, various tests can be performed on the A cells and the B cells, including cell morphology, cell number, expression of genes and proteins in the cells, and the like.
Example 2
This example is basically the same as example 1 except that the microporous layer 18 is located in the middle of the a-type culture dish body 3 as shown in fig. 6. This arrangement allows not only to test the interaction of two cells without contact, as in example 1, but also to test the interaction of two cells with contact.
The specific embodiment is as follows:
the a-type culture dish is fixed on the lifting beam 8, and the lifting beam 8 is rotated so that the a-type culture dish is not directly above the B-type culture dish 2. The a cells are inoculated into the a-type culture dish body 3, then only the culture medium (not inoculated cells) is added to the B-type culture dish 2, then the handle 24 is turned to descend the a-type culture dish, and when the a-type culture dish descends to the B-type culture dish 2, the lugs 6 of the a-type culture dish contact the upper surface of the B-type culture dish 2, the turning of the handle 24 is stopped. Then, the culture dish cover 17 is covered for cell culture, after the A cells are grown on the wall, the handle 24 is rotated, and the lifting beam 8 is enabled to descend, so that the A-type culture dish body 3 is separated from the sealing disc 4. Then the culture dish cover 17 is taken out, the lifting beam 8 is lifted by rotating the handle 24, the A-type culture dish body 3 is supported by the sealing disc 4, and the A-type culture dish body 3 is taken out from the culture tank 15. The medium in the A-type culture dish body 3 and the B-type culture dish 2 was aspirated with a pipette, and then the A-type culture dish body 3 was inverted on the sealing plate 4, and B cells were seeded in the other vessel of the A-type culture dish body 3. As shown in fig. 6, a cell a is first inoculated into the cavity above the type a dish body 3, then the type a dish body 3 is inverted, and a cell B is inoculated into the original cavity below. After inoculation is completed, fresh culture medium is added into the culture tank 15 of the type-B culture dish 2, the position of the hanging lugs 6 is adjusted, then the handle 24 is rotated to enable the type-A culture dish body 3 to descend, after the hanging lugs 6 are contacted with the upper surface of the type-B culture dish 2, the handle 24 is continuously rotated to enable the sealing disc 4 to descend, and the type-A culture dish is suspended in the culture tank 15. And ensures that microporous layer 18 is below the level of the medium in culture tank 15 to ensure that the a cells are completely infiltrated by the medium. Cell-to-cell direct contact (e.g., partial cell membrane contact) occurs between the a cells and the B cells through the micropores in microporous layer 18. After a period of culture, the A-type culture dish body 3 is taken out, and various tests can be carried out on the A cells and the B cells, including cell morphology, cell number, expression of genes and proteins in the cells and the like.
Example 3
The difference of this embodiment is that, as shown in fig. 7, a partition board 28 perpendicular to the microporous layer 18 is provided in the a-type culture dish body 3, the partition board 28 divides the cavity of the a-type culture dish body 3 into two chambers, and the partition board 28 is sealed and adhered to the microporous layer 18 and the side wall of the a-type culture dish body 3 respectively. By adopting the scheme, the A cells and the C cells can be respectively inoculated in the two chambers of the A-type culture dish body 3, the B cells can be inoculated in the B-type culture dish 2, and the co-culture condition of the three cells can be observed and tested.
The foregoing is merely exemplary embodiments of the present invention, and specific structures and features that are well known in the art are not described in detail herein. It should be noted that modifications and improvements can be made by those skilled in the art without departing from the structure of the present invention, and these should also be considered as the scope of the present invention, which does not affect the effect of the implementation of the present invention and the utility of the patent. The protection scope of the present application shall be subject to the content of the claims, and the description of the specific embodiments and the like in the specification can be used for explaining the content of the claims.
Claims (10)
1. The cell co-culture equipment is characterized by comprising a type B culture dish, a type A culture dish and a lifting unit; the A-type culture dish comprises an A-type culture dish body, a sealing disc, a hanging lug and a vertical rod, wherein a microporous layer is fixed in the A-type culture dish body, and the periphery of the microporous layer is fixedly connected with the inner wall of the A-type culture dish body in a sealing manner; the hanging lugs are fixed on the A-type culture dish body, the number of the hanging lugs is a plurality, and the hanging lugs are in axisymmetric distribution by taking the central axis of the A-type culture dish body as a symmetry axis; the sealing disc is used for sealing the lower end of the A-type culture dish body, the lower end of the A-type culture dish body is clamped and fixed with the sealing disc, the sealing disc is integrally formed with the vertical rod, and the vertical rod is fixed on the lifting unit; the B-type culture dish comprises a culture tank which is positioned right below the A-type culture dish body, and the hanging lugs are used for hanging the A-type culture dish body in the culture tank; the lifting unit is used for driving the vertical rod to move in the vertical direction, the vertical rod is used for applying acting force to the sealing disc, so that the sealing disc is separated from the A-type culture dish body, and the A-type culture dish body is taken out of the culture tank of the B-type culture dish through the sealing disc.
2. The cell co-cultivation apparatus according to claim 1, wherein said microporous layer is a polycarbonate membrane.
3. A cell co-cultivation apparatus according to claim 2, wherein said microporous layer has a microporous diameter of 0.5-8 μm.
4. A cell co-cultivation apparatus according to claim 3, wherein the number of culture tanks of the type B dishes is a number corresponding to the number of culture tanks.
5. The apparatus according to claim 4, wherein the culture vessel is a cylindrical vessel with an upper opening, and the body of the culture vessel is cylindrical.
6. The cell co-culture apparatus according to claim 5, wherein two sets of ear fixing groove groups for inserting the ear are provided on the outer side of the side wall of the A-type culture dish body, the ear fixing groove groups comprise a plurality of ear fixing grooves, the ear fixing grooves are distributed along the axial direction of the A-type culture dish body, and the two sets of ear fixing grooves are distributed in an axisymmetric manner by taking the center axis of the A-type culture dish body as a symmetry axis.
7. The cell co-culture apparatus of claim 6, wherein the lifting unit comprises a lifting beam, a screw mechanism, and a base; the vertical rod is detachably fixed on the lifting beam, the base is used for fixing a screw rod mechanism and placing a B-type culture dish, and the screw rod mechanism is used for lifting the lifting beam; the lifting beam is horizontally arranged and is rotationally connected with the screw rod mechanism.
8. The cell co-culture apparatus of claim 7, wherein the base is provided with a placement groove for placing a type B culture dish.
9. The cell co-culture apparatus according to claim 8, wherein a partition plate is provided in the body of the type A culture dish, the partition plate is perpendicular to the microporous layer, and the partition plate is used for dividing the cavity of the body of the type A culture dish into a plurality of chambers.
10. The cell co-culture apparatus of claim 9, wherein the microporous layer is located in the middle of the body of the type a plate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911141962.8A CN110804548B (en) | 2019-11-20 | 2019-11-20 | Cell co-culture equipment |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911141962.8A CN110804548B (en) | 2019-11-20 | 2019-11-20 | Cell co-culture equipment |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110804548A CN110804548A (en) | 2020-02-18 |
| CN110804548B true CN110804548B (en) | 2023-05-30 |
Family
ID=69490723
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911141962.8A Active CN110804548B (en) | 2019-11-20 | 2019-11-20 | Cell co-culture equipment |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110804548B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103194388A (en) * | 2013-03-22 | 2013-07-10 | 中国科学院力学研究所 | Double-layer cell co-culture device with adjustable gap |
| CN103270149A (en) * | 2010-10-08 | 2013-08-28 | 莱萃恩维斯柯凡股份有限公司 | Cell culture insert |
| CN204737966U (en) * | 2015-05-28 | 2015-11-04 | 深圳职业技术学院 | Multiple cell culture apparatus |
| CN206385177U (en) * | 2017-01-10 | 2017-08-08 | 中国人民解放军第四军医大学 | The transwell culture dishes co-cultured suitable for three kinds of cells |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104169413B (en) * | 2012-02-29 | 2016-01-20 | 株式会社Jtec | There is the cell culture apparatus that nutrient solution changes function |
-
2019
- 2019-11-20 CN CN201911141962.8A patent/CN110804548B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103270149A (en) * | 2010-10-08 | 2013-08-28 | 莱萃恩维斯柯凡股份有限公司 | Cell culture insert |
| CN103194388A (en) * | 2013-03-22 | 2013-07-10 | 中国科学院力学研究所 | Double-layer cell co-culture device with adjustable gap |
| CN204737966U (en) * | 2015-05-28 | 2015-11-04 | 深圳职业技术学院 | Multiple cell culture apparatus |
| CN206385177U (en) * | 2017-01-10 | 2017-08-08 | 中国人民解放军第四军医大学 | The transwell culture dishes co-cultured suitable for three kinds of cells |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110804548A (en) | 2020-02-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| AU2006257609B2 (en) | Cell-and tissue culture device | |
| JP5558560B2 (en) | Bioreactor system | |
| RU2016152238A (en) | METHODS OF PERFUSION CULTIVATION AND THEIR APPLICATION | |
| US3912596A (en) | Multiple inoculating system | |
| WO2003085080A1 (en) | Cell culture plate and system using the same | |
| US4968623A (en) | Cell culture apparatus | |
| CN110804548B (en) | Cell co-culture equipment | |
| CN203212573U (en) | Device for improving culturability of microorganisms | |
| CN113846016A (en) | High-flux porous array chip, device, preparation method and application | |
| US20100028992A1 (en) | Cell culture device | |
| CN109929762A (en) | A kind of cell culture apparatus | |
| CN115948339A (en) | Method for culturing glioma organoid by high-flux gas-liquid boundary method | |
| US4144136A (en) | Apparatus for cellular culture | |
| CN204644372U (en) | A kind of many cells noncontact culture apparatus | |
| CN115612615A (en) | Culture dish convenient for observation for biological cell culture | |
| KR20190068378A (en) | Culture device of inserting type for three dimensional cell culture, kit and co-culture method of three dimensional cell using the same | |
| CN210528952U (en) | Multi-functional incubator of constant temperature cell | |
| CN203429184U (en) | Multi-adherent-cell coculture device | |
| CN220413416U (en) | Organoid culture bracket and culture plate for culturing organoid by using matrigel | |
| CN215668045U (en) | Cell vessel for cell culture | |
| CN212247075U (en) | Cell culture device is used in medical science laboratory | |
| CN215593084U (en) | Culture plate plug-in, single-hole culture plate and multi-hole culture plate | |
| CN215799463U (en) | Cell material inoculation fixer | |
| CN210163461U (en) | A culture apparatus for cell proliferation | |
| CN220780403U (en) | Cell culture bottle bracket |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |