CN108611273A - Cell co-cultures unit - Google Patents
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- CN108611273A CN108611273A CN201810386179.7A CN201810386179A CN108611273A CN 108611273 A CN108611273 A CN 108611273A CN 201810386179 A CN201810386179 A CN 201810386179A CN 108611273 A CN108611273 A CN 108611273A
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- 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
- C12M23/22—Transparent or translucent parts
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Abstract
The present invention provides cells to co-culture unit, belong to technical field of cell culture, cell co-cultures the culture dish that unit includes at least two settings stacked on top of one another, two culture dishes of arbitrary neighborhood include upper culture dish and lower culture dish, the interior circulation passage of both connections inner cavity or interior seepage channel are equipped between upper culture dish and lower culture dish, the upper culture dish is equipped with the upper bridging arrangement being overlapped on the lower culture dish, and the lower part of upper culture dish is inserted into lower culture dish, the upper bridging arrangement is equipped with the upper ventilating structure for making the inner cavity of the lower culture dish in adjacent two culture dish be in communication with the outside.Different cells is cultivated on each culture dish to simulate the growing environment of normal body inner cell, and stacks mode using mutually overlapped and co-cultures building for unit convenient for the cell, and it is inconvenient to solve the problems, such as that cell in the prior art co-cultures unit operation.
Description
Technical field
The present invention relates to cells to co-culture unit.
Background technology
To using traditional single layer face cell culture in current external toxicity and pharmacological experiment, which can not
Internal microenvironment residing for true simulation cell, certain deviation may be caused when Study of Exogenous substance is to matrix effect.
In order to preferably establish the cultivating system similar to vivo environment, the late 1980s, researcher is single in original two dimension
Cell Co culturing Techenique has been developed on the basis of confluent monolayer cells culture.
Cell Co culturing Techenique is the technology in unified environment by two or more cell co-culture, comparison
For unicellular monolayer cultivation, have the advantages that preferably to react vivo environment.It is directly connect currently, existing co-cultivation mode has
It touches to co-culture and co-cultures two kinds with mediate contact, although being in direct contact the state for co-culturing and can solving various kinds of cell co-cultivation,
The culture but cell for being directed to different tissue sources mixes, independent cannot be distinguished, different cells phase in growth course
Mutually contact interacts, and makes the expression of the respective function of the cell to mix or the character of various cells is caused to change
Become, for the later stage study cell influence each other and when respective effect, Chang Wufa distinguishes or loses actual response；It uses at present
Mediate contact co-cultivation be usually directed to two kinds of cells, most three kinds of cells, although good area can be carried out between cell
Point, but due to by current co-culture device and technical limitation, usually only co-culturing three kinds of cells, in addition extracellular training
Foster base is usually stationary state, these all simulate Cell culture invitro in good cell and are produced in the internal environment of body as far as possible
Certain obstacle is given birth to.
Chang Yan, Wei Wei deliver entitled in July, 2009 in《Cell co-cultures and its progress of application》Summary,
In refer to a kind of Millicell plug-in types Tissue Culture Dish（The also known as cells Transwell）, when carrying out cell co-cultivation,
The cells Transwell are put into culture plate, are planted in upper chamber and lower room respectively into cell A and cell B, and upper chamber bottom is set
The film with permeability of upper chamber and lower room can be connected to by having, and make composition influences of the cell B in its culture solution to cell A,
The influence of materials onto cells A growths, movement to study cell B secretions or metabolism generation etc..But the cells Transwell
The mode of building avoid cumbersome, make troubles to the operation of experimenter.
Invention content
The purpose of the present invention is to provide a kind of cells to co-culture unit, and list is co-cultured to solve cell in the prior art
The problem of atom operation inconvenience.
To achieve the above object, the technical solution of cell of the invention co-cultivation unit is：
Cell co-cultures the culture dish that unit includes at least two settings stacked on top of one another, and two culture dishes of arbitrary neighborhood include upper training
Ware and lower culture dish are supported, interior circulation passage or the interior seepage flow that both connections inner cavity is equipped between upper culture dish and lower culture dish are logical
Road, it is characterised in that：The upper culture dish, which is equipped with, is overlapped on upper bridging arrangement on the lower culture dish, and upper culture dish
Lower part is inserted into lower culture dish, the upper bridging arrangement be equipped with for make the inner cavity of the lower culture dish in adjacent two culture dish with
The upper ventilating structure of external world's connection.
The beneficial effects of the invention are as follows：Culture dish is stacked, and cultivate on each culture dish different cells with
The growing environment of normal body inner cell is simulated, and stacks mode using mutually overlapped and co-cultures unit convenient for the cell
It builds, it is inconvenient to solve the problems, such as that cell in the prior art co-cultures unit operation.
Further, it overlaps for convenience, the opening's edge of the lower culture dish in two adjacent culture dishes is formed and corresponding upper training
The supporting mouth edge for supporting the upper bridging arrangement support cooperation of ware, by overlapping the culture dish for being in upper layer in two adjacent culture dishes
At the opening's edge of the culture dish in lower layer, convenient for stacking between culture dish.
Further, in order to improve the safety that cell co-cultures unit, on the upper culture dish in two adjacent culture dishes
Upper bridging arrangement include the overhead gage for circumferentially extending or being distributed along upper culture dish, the upper ventilating structure is arranged in the upper gear
On plate, the culture in lower layer is overlapped on by the way that overhead gage is arranged on the culture dish in upper layer in two adjacent culture dishes
On ware, reduce the device stacks height, improves safety.
Further, cell co-cultures unit and is additionally provided with head cover, and cell, which co-cultures, is in undermost culture dish in unit
It is equipped with for head cover overlapped lower bridging arrangement, and head cover and cell co-culture the opening's edge of the culture dish in top layer in unit
Place interval is arranged, and the lower ventilating structure being additionally provided on the head cover and/or lower bridging arrangement inside and outside connection head cover avoids cell total
The hypercellularity turned out in culture unit contacts the infection for leading to cell with air.
Further, in order to facilitate stacking between culture dish, the downside of the overhead gage is equipped under accordingly
The restriction protrusion knot that the supporting mouth of culture dish coordinates along arrangement and with the internal face of lower culture dish and/or the limit of outside wall surface block
Structure is limited by the block on restriction protrusion structure and supporting mouth edge and is coordinated, keeps the upper culture dish in two adjacent culture dishes concentric
It overlays on lower culture dish, improves the stability that cell co-cultures unit.
Further, stacking in order to facilitate culture dish, the upper bridging arrangement of upper culture dish includes in adjacent two culture dish
It is sleeved on the supporter of corresponding lower culture dish outer circumferential, the lower support that the lower end of the supporter is equipped with corresponding lower culture dish
Structure top pressure coordinates, and is coordinated by supporter and lower support structure top pressure to support upper culture dish in adjacent two culture dish.
Further, the cell co-cultures multiple culture dishes that unit includes at least three arrangements stacked on top of one another successively,
Multiple culture dishes include top culture dish, bottom culture plate and at least one intermediate culture dish, outside at least one intermediate culture dish
Week is installed with medial support structures, which forms upper overlap joint of the intermediate culture dish as the upper culture dish when
Structure, and form lower support structure of the intermediate culture dish as the lower culture dish when.
Further, the upper bridging arrangement of the top culture dish includes overhead gage and is arranged at the opening's edge of overhead gage simultaneously
The support ring flat-plate to overhang downwards, facilitates processing.
Further, the medial support structures of intermediate culture dish include overhead gage and be arranged at the opening's edge of overhead gage and to
The support ring flat-plate of lower overhanging, facilitates processing.
Further, the bottom culture plate periphery is installed with lower supporting baffle, which forms bottom culture
Lower support structure when ware is as the lower culture dish, it includes top of the spiral-lock on the culture dish that the cell, which co-cultures unit,
Lid, there are ventilative gap between head cover and the opening's edge of the top culture dish, the lower end support of the head cover be placed on it is described under
In supporting baffle, avoid the hypercellularity turned out in cell co-cultivation unit contacts the infection for leading to cell with air.
Further, the supporter be support ring set, the lower support structure be equipped with restriction protrusion structure with branch
The internal face of pushing out ring set and/or the limit cooperation of outside wall surface block, avoid stacking dislocation between culture dish.
Further, the supporter is multiple support columns of circumferentially spaced arrangement, and multiple support columns coordinate shape
Circumferential surface is positioned at interior positioning circumferential surface and/or outside, the lower support structure is equipped with restriction protrusion structure with all with the interior positioning
Face and/or outer positioning circumferential surface block limit cooperation avoid stacking dislocation between culture dish.
Further, for the ease of operating, the overhead gage of the upper culture dish in two adjacent culture dishes is equipped with for liquid relief
Rifle is inserted into so as to the connecting hole for descending culture dish adding liquid accordingly, the connecting hole constitutes the upper ventilating structure, passes through
The connecting hole of overhead gage, which can be opposite in the culture dish on the downside of it, adds nutrient solution, digestive juice or contamination agent etc., without needing
Cell is dismounted when adding liquid into culture dish and co-culture unit.
Further, in order to improve air permeability effect, the connecting hole on same overhead gage was equipped at least two and along week
To arrangement.
Further, for the structure of simplified culture ware, the bottom wall of the upper culture dish in two adjacent culture dishes is equipped with
The interior seepage channel.
Further, in order to reduce the manufacture difficulty of culture dish, the bottom wall of culture dish is gone up in two adjacent culture dishes by oozing
Permeable membrane is formed, and the micropore on permeable membrane constitutes the interior seepage channel, without carrying out multi-processing to culture dish.
Further, in order to keep cell preferably adherent, helpful cell patch is all provided on the inside bottom wall of each culture dish
Wall helps leapfrog structure.
Further, for the ease of the operation of experimenter, the cell co-culture unit be equipped with for outer circulation
The entrance and exit of pipeline connection can be co-cultured unit by outer runner pipe road direction cell and supply nutrient solution or discharge nutrition
Liquid is conveniently operated.
Further, the arrangement of outer flow pipe, the entrance and exit are set at undermost training for convenience
It supports on ware, so that outer flow pipe is arranged lower, be convenient for the setting of outer flow pipe.
Description of the drawings
Fig. 1 is the structural schematic diagram that the cell of the present invention co-cultures the specific embodiment 1 of unit；
Fig. 2 is the front view for the bottom culture plate that the cell of the present invention co-cultures the specific embodiment 1 of unit；
Fig. 3 is the vertical view for the bottom culture plate that the cell of the present invention co-cultures the specific embodiment 1 of unit；
Fig. 4 is the front view for the intermediate culture dish that the cell of the present invention co-cultures the specific embodiment 1 of unit；
Fig. 5 is the vertical view for the intermediate culture dish that the cell of the present invention co-cultures the specific embodiment 1 of unit；
Fig. 6 is the front view for the top culture dish that the cell of the present invention co-cultures the specific embodiment 1 of unit；
Fig. 7 is the vertical view for the top culture dish that the cell of the present invention co-cultures the specific embodiment 1 of unit；
Fig. 8 is the axonometric drawing for the head cover that the cell of the present invention co-cultures the specific embodiment 1 of unit；
Fig. 9 is the front view for the head cover that the cell of the present invention co-cultures the specific embodiment 1 of unit；
Figure 10 is the axonometric drawing that the cell of the present invention co-cultures that the chain of the specific embodiment 1 of unit is taken over；
Figure 11 is the structural schematic diagram that the cell of the present invention co-cultures the specific embodiment 2 of unit；
Figure 12 is the front view for the bottom culture plate that the cell of the present invention co-cultures the specific embodiment 2 of unit；
Figure 13 is the front view for the intermediate culture dish that the cell of the present invention co-cultures the specific embodiment 2 of unit；
Figure 14 is the axonometric drawing for the intermediate culture dish that the cell of the present invention co-cultures the specific embodiment 2 of unit；
Figure 15 is the vertical view for the intermediate culture dish that the cell of the present invention co-cultures the specific embodiment 2 of unit；
Figure 16 is the axonometric drawing for the intermediate culture dish that the cell of the present invention co-cultures the specific embodiment 3 of unit.
Specific implementation mode
Embodiments of the present invention are described further below in conjunction with the accompanying drawings.
The cell of the present invention co-cultures the specific embodiment 1 of unit, and as shown in Figures 1 to 10, cell co-cultures unit packet
A pipeline 5 flowed for nutrient solution is included, peristaltic pump 6 is in series on pipeline 5 and two cells co-cultured for cell are trained altogether
Unit is supported, drives nutrient solution to be flowed in pipeline 5 by peristaltic pump 6 and does not generate retardance and destroy the effect of cell, is sought
Nutrient solution is co-cultured at two to be flowed in unit, and the metabolism product of cell is taken away while for cells with nutrient, and
The flowing of the blood of the flow simulating body of nutrient solution, especially for for the cell of different tissues organ generally in body
All it is to be transmitted into row information by blood and mass exchange, therefore can preferably simulates micro-loop of the cell in body
Border.Wherein, peristaltic pump 6 constitutes the pumping installations for driving the flowing of the nutrient solution in each culture dish.
In the present embodiment, cell co-cultures unit and includes about three spaced apart culture dishes and tip upside down on three cultures
Head cover 4 on ware, wherein three culture dishes are respectively top culture dish 3, intermediate culture dish 2 and bottom culture plate 1 from top to bottom,
It therefore overlaps between three culture dishes, and top culture dish to reduce the difficulty built when cell co-cultures unit
3, the size of intermediate culture dish 2 and bottom culture plate 1 is sequentially increased, top culture dish 3, intermediate culture dish 2 and bottom culture plate 1
Gusset on be respectively equipped with top annular baffle 301, intermediate annular baffle 201 and base circle baffle 101, wherein top annular
The downside of baffle 301 and the opening's edge overlap joint of intermediate culture dish 2 coordinate, and downside and the bottom of intermediate annular baffle 201 are cultivated
Cooperation is overlapped at the opening's edge of ware 1, base circle baffle 101 and the opening's edge overlap joint of head cover 4 coordinate, and top annular baffle 301
Race diameter be less than the race diameter of intermediate annular baffle 201, the race diameter of intermediate annular baffle 201 is less than base circle
The race diameter of baffle 101, in addition there are the vertical of 1-2mm between the inside bottom wall of head cover 4 and the opening's edge of top culture dish 3
Then head cover 4 is tipped upside down on bottom part ring by interval, the permeability for improving top culture dish 3 by the overlap joint successively of three culture dishes
On shape baffle 101, so that cell is co-cultured unit and be easier to assemble, be convenient for the operation of experimenter.Wherein, top annular baffle
301 and intermediate annular baffle 201 constitute overhead gage, base circle baffle 101 constitute lower baffle plate.
Bottom culture plate 1 is equipped with the entrance 103 being connect with pipeline 5 and outlet 104, distinguishes in entrance 103 and outlet 104
It is threadedly coupled with pipeline 5, and chain take over 7 is connected between the bottom culture plate 1 of two cells co-cultivation units, chain take over 7
Material, which is transparent material and chain take over 7, has certain elasticity, can achieve the effect that sealing, one end of chain take over 7 with
The outlet 104 that one of cell co-cultures the bottom culture plate 1 of unit is threadedly coupled, the other end of chain take over 7 and another
Cell co-culture unit bottom culture plate 1 entrance 103 be threadedly coupled, and chain take over 7 can with two bottom culture plates 1 it
Between realize sealing cooperation, avoid outside air pollute nutrient solution.Bottom culture plate 1, intermediate culture dish 2 and top culture dish 3
Be coated on the bottom wall of inside help cell adherent help leapfrog structure 102, the coating of poly-D-lysine material is selected in the present embodiment
It helps cell adherent on the inside bottom wall of bottom culture plate 1, and is equipped with support bottom below the outside bottom wall of bottom culture plate 1
The support construction of portion's culture dish 1, when making bottom culture plate 1 be placed on to be operated on workbench, the entrance of bottom culture plate 1 and
It keeps being spaced between outlet and workbench, it is easy to operation convenient for the docking of the entrance and outlet of chain take over 7 and bottom culture plate 1.
Certainly, in other embodiments, it so that the entrance and exit of bottom culture plate 1 is higher than the bottom wall of bottom culture plate 1, be convenient for chain
The docking of the entrance and outlet of take over 7 and bottom culture plate 1.Wherein, pipeline 5 and 7 composition of chain take over flow outer for nutrient solution
Flow pipe.
The inside bottom wall of intermediate culture dish 2 and top culture dish 3 is pvdf membrane, wherein the pvdf membrane of intermediate culture dish 2
For intermediate pvdf membrane 202, the pvdf membrane of top culture dish 3 is top pvdf membrane 302 can make certainly in other embodiments
Cell growth and there is certain pore size to be all possible convenient for the material of mass exchange, such as film class, hydrogel or 3D micropore classes
Material etc..Wherein, the inside bottom wall of culture dish constitutes the cell inoculation face of culture dish, and pvdf membrane constitutes permeable membrane, on pvdf membrane
Micropore constitute and be connected to intracavitary seepage channel in two neighboring culture dish, and top culture dish 3, intermediate culture dish 2 and bottom training
The cell inoculation face for supporting ware 1 is placed in the lower section at the opening's edge of bottom culture plate 1, enable nutrient solution submerge be placed in it is same thin
Born of the same parents co-culture three cell inoculation faces of unit.It certainly, in other embodiments, can be by the way that pipeline be arranged by adjacent culture
It is connected between ware, micropore can also be opened up on the inside bottom wall of intermediate culture dish 2 and top culture dish 3, bottom is cultivated
It is connected to carry out mass exchange and information between ware 1 and intermediate culture dish 2, between top culture dish 3 and intermediate culture dish 2
It transmits.
Preferably, the distance between opening's edge of the base circle baffle 101 away from bottom culture plate 1 is equal to bottom culture plate 1
Opening's edge to bottom culture plate 1 the distance between bottom wall 1/5th, intermediate annular baffle 201 is away from intermediate culture dish 2
Opening's edge between distance be equal to five points of the distance between bottom wall at the opening's edge of intermediate culture dish 2 to intermediate culture dish 2
One of, the distance between the bottom wall of top annular baffle 301 away from top culture dish 3 is equal at the opening's edge of top culture dish 3 to top
/ 5th of the distance between the bottom wall of portion's culture dish 3 reduce the height that cell co-cultures unit, improve safety as far as possible
Property.
Preferably, it has circumferentially been evenly arranged eight connections respectively on intermediate annular baffle 201 and top annular baffle 301
Hole, wherein the connecting hole being placed on intermediate annular baffle 201 is intermediate connecting hole 203, the company being placed on top annular baffle 301
It is top connecting hole 303 to connect hole, convenient for culture medium, nutrient solution or contamination liquid etc., and intermediate ring are added in subsequent operation
Intermediate connecting hole 203 on shape baffle 201 makes to keep connection between bottom culture plate 1 and head cover 4, on top annular baffle 301
Top connecting hole 303 make to keep connection between intermediate culture dish 2 and head cover 4, ensure bottom culture plate 1 and intermediate culture dish 2
Permeability, and head cover 4 is directly overlapped on base circle baffle 101 and understands some gas and enter in head cover 4, avoids cell
Because dead for hypoxgia.Gap between head cover 4 and base circle baffle 101 constitutes lower ventilative inside and outside connection head cover 4
Structure, certainly, in other embodiments, it is contemplated that cell co-cultures the permeability of unit, ensures the eupnea of cell, because
This, several grooves can also be arranged in ventilating structure at the opening's edge of head cover 4, or can also be set on base circle baffle 101
Air hole is set, to improve the air inlet in head cover 4.It can also be arranged in the lap-joint of head cover 4 and base circle baffle 101
Air-permeable paper, or make head cover 4 equipped with several fulcrums at lap position at the opening's edge of base circle baffle 101 and head cover 4
Certain interval is kept to ensure permeability in turn between base circle baffle 101.Wherein, intermediate connecting hole 203 and top connect
It connects hole 303 to constitute for liquid-transfering gun insertion with the connecting hole to the culture dish adding liquid in lower layer, connecting hole is constituted for making
The upper ventilating structure that the inner cavity of culture dish in lower layer is in communication with the outside.Certainly, in other embodiments, upper ventilating structure can
The fan groove in intermediate annular baffle 201 and top annular baffle 301 is arranged.
Preferably, an isocon can also be connected on chain take over 7, it can be from this implementation of one end of isocon supplement liquid
In example, isocon is four-way isocon, and two are connected to the chain take over 7 respectively with culture dish, remaining is two in, and one to for being discharged
Cell co-cultures the liquid recycled in unit, another to can fill in rubber stopper in oral area, liquid or remove liquid when
Time can be inserted through rubber belt by syringe needle and carry out liquid supplement or absorption.
In the present embodiment, in order to make to stack more stable, top annular baffle 301 and intermediate annular baffle between culture dish
The upper limit block of the opening's edge arrangement of the opening's edge and bottom culture plate 1 around intermediate culture dish 2 is respectively equipped on 201 downside
304 and middle limit block 204, wherein upper limit block 304 and the internal face block limit of the opening's edge of intermediate culture dish 2 coordinate,
Middle limit block 204 coordinates with the internal face block limit at the opening's edge of bottom culture plate 1, is trained by bottom culture plate 1, centre
It supports when being stacked successively between ware 2 and top culture dish 3, passes through middle limit block 204 and the inner wall at the opening's edge of bottom culture plate 1
The block between the internal face at block limit and the opening's edge of upper limit block 304 and intermediate culture dish 2 between face limits, and makes
It can be stacked with one heart between culture dish, improve cell and co-culture the stability of unit, and can played and certain prevent from cultivating
The problem of cell contamination in ware.Wherein, upper limit block 304 and middle limit block 204 are constituted around corresponding lower culture dish
The restriction protrusion structure that supporting mouth coordinates along arrangement and with the internal face of lower culture dish and/or the limit of outside wall surface block, certainly,
In other embodiment, upper limit block 304 and middle limit block 204 can also respectively with it is outer at the opening's edge of intermediate culture dish 2
Side, bottom culture plate 1 opening's edge at lateral surface block limit cooperation；Restriction protrusion structure may be limited post or limit
The structures such as position ring set, meanwhile, it is also unlimited for upper limit block 304 and 204 quantity of middle limit block, two can be arranged
A, three, any amounts such as four.
When being ready for cell co-cultivation operation, cell is first co-cultured to the bottom culture plate 1 and pipeline 5, chain of unit
Take over 7, peristaltic pump are sequentially connected, then cell inoculation is co-cultured to the thin of the bottom culture plate 1 of unit in two cells again
On born of the same parents' inoculating surfaces, nutrient solution is added in bottom culture plate 1 followed by liquid-transfering gun, slight concussion and etc. after make cell
Suspension is uniformly distributed in bottom culture plate 1, then in the same fashion puts intermediate culture dish 2, top culture dish 3 successively
It sets on bottom culture plate 1, if the liquid level of nutrient solution can pass through intermediate connecting hole 203 and top connecting hole when inadequate
303 addition nutrient solutions make the cell inoculation face of top culture dish 3 be placed in nutrition so that nutrient solution reaches certain liquid level
Under the liquid level of liquid, then head cover 4 is covered, is put into cell culture in incubator.It waits after the completion of cultivating, you can subsequently tried
It tests.Certainly, in other embodiments, peristaltic pump can not be connected in series on pipeline 5 when cell co-cultures, bottom culture plate 1,
Pipeline 5 still keeps being connected to chain take over 7, and the liquid in bottom culture plate is avoided to be flowed out from bottom culture plate.
It is co-cultured with pneumonocyte, vascular endothelial cell, macrophage and liver cell to study transpulmonary contamination in the present embodiment
There are related inflammation caused by liver metabolism or metabolic effect.Wherein it is inoculated with pneumonocyte, vascular endothelial cell and macrophage
Cell co-cultures unit simulation organ lung, and the cell for being inoculated with liver cell, vascular endothelial cell and macrophage co-cultures unit
Simulated organ liver.
When inoculating cell, one of cell co-cultures and is inoculated with macrophage on the bottom culture plate 1 in unit, centre training
It supports and is inoculated with vascular endothelial cell on ware 2, pneumonocyte is inoculated in top culture dish 3, another cell co-cultures the bottom in unit
Culture dish 1 is inoculated with macrophage, is inoculated with vascular endothelial cell in intermediate culture dish 2, liver cell, mould are inoculated in top culture dish 3
The quasi- cyclic process that liver is normally flow to by lung channel intravascular mass.When the lung being inoculated in the culture dish of pneumonocyte is thin
When born of the same parents contaminate, macrophage can the migration in each culture dish under the driving of peristaltic pump, the flowing of blood in similar blood vessel,
Detect the variation of the cell survival and associated metabolites of the relevant culture dish for being inoculated with liver cell, or research inflammatory factor water
The migration of flat correlated expression and macrophage.
Meanwhile unit is co-cultured using the cell and carries out exposure experiment, contamination object can be made to pass sequentially through top culture dish
In pneumonocyte, the vascular endothelial cell of intermediate culture dish, bottom culture plate macrophage, simulation body really contaminated
Journey more meets true cell toxic environment in body.
Certainly, in other embodiments, any cell can co-culture unit to cultivate by the cell, if needed
Carry out co-culturing more cells only needs intermediate culture dishes 2 to be arranged or more cells of connecting on pipeline 5 co-culture more
Unit, in addition, the cell co-cultivation unit of the present invention also can individually be connected on pipeline 5 and use.
The cell of the present invention co-cultures the specific embodiment 2 of unit, and the specific reality of unit is co-cultured with the cell of the present invention
Example 1 is applied to differ only in：As shown in Figure 11 to Figure 15, the outer edge of top annular baffle 31 and intermediate annular baffle 21 it is outer
The top braces ring set 32 to overhang downwards and intermediate supports ring set 22, the lower end top of top braces ring set 32 are respectively equipped on edge
It is pressed on the upper side of intermediate annular baffle 21, the lower end top pressure of intermediate supports ring set 22 is on base circle baffle 11, wherein
Top braces ring set 32 and intermediate supports ring set 22 constitute support ring set, and it is circumferential that support ring set composition is sleeved on accordingly lower culture dish
The supporter in outside, top annular baffle 31 and top braces ring set 32 constitute the upper bridging arrangement of top culture dish, intermediate ring
Shape baffle 21 and intermediate supports ring set 22 constitute the medial support structures of intermediate culture dish, and base circle baffle 11 constitutes lower support
Baffle.In the present embodiment, between being respectively equipped with along the circumferential direction on the upper side of intermediate annular baffle 21 and base circle baffle 11
Every the multiple realizing middle limiting columns 23 and bottom limited post 13 of arrangement, pass through the outer wall of realizing middle limiting column 23 and top braces ring set 32
Limit block coordinates between face and/or internal face, the outside wall surface and/or internal face of bottom limited post 13 and intermediate supports ring set 22
Between limit block coordinate, make to be stacked together with one heart between culture dish, 13 structure of realizing middle limiting column 23 and bottom limited post
At restriction protrusion structure.Certainly, in other embodiments, restriction protrusion structure may be to be arranged in 21 He of intermediate annular baffle
Limit ring set on the upper side of base circle baffle 11；The setting quantity of realizing middle limiting column and bottom limited post is unlimited simultaneously
, two, three, any amounts such as four can be set.
In the present embodiment, top annular baffle 31, intermediate annular baffle 21 and base circle baffle 11 are equipped with handle 8,
The hand-held of personnel easy to operation, certainly, in other embodiments, handle 8 may also be arranged on top culture dish, intermediate culture
In the outside wall surface of ware and bottom culture plate.
The cell of the present invention co-cultures the specific embodiment 3 of unit, and the specific reality of unit is co-cultured with the cell of the present invention
Apply differing only in for example 1：As shown in figure 16, supporter may include outer edge and the top for being separately positioned on intermediate annular baffle
The outer edge of portion's ring baffle and the multiple overlap joint columns 9 to overhang downwards.
The cell of the present invention co-cultures the specific embodiment 4 of unit, and the specific reality of unit is co-cultured with the cell of the present invention
Apply differing only in for example 1：Intermediate culture dish is arranged two, and the diameter of two intermediate culture dishes is sequentially increased from top to bottom.
The cell of the present invention co-cultures the specific embodiment 5 of unit, and the specific reality of unit is co-cultured with the cell of the present invention
Apply differing only in for example 1：The peripheral surface of the culture dish in upper layer is equipped with and is in for being overlapped in adjacent culture dish
Overlap joint ear at the opening's edge of the culture dish of lower layer.
The cell of the present invention co-cultures the specific embodiment 6 of unit, and the specific reality of unit is co-cultured with the cell of the present invention
Apply differing only in for example 1：Bottom culture plate is taken over pipeline, chain and is connect by hose clamp or anchor ear.
Claims (19)
1. cell co-cultures unit, include the culture dish of at least two settings stacked on top of one another, two culture dishes of arbitrary neighborhood include
Upper culture dish and lower culture dish are equipped with the interior circulation passage of both connections inner cavity or interior seepage flow between upper culture dish and lower culture dish
Channel, it is characterised in that：The upper culture dish is equipped with the upper bridging arrangement being overlapped on the lower culture dish, and upper culture dish
Lower part be inserted into lower culture dish, the upper bridging arrangement is equipped with the inner cavity for making the lower culture dish in adjacent two culture dish
The upper ventilating structure being in communication with the outside.
2. cell according to claim 1 co-cultures unit, it is characterised in that：Lower culture dish in two adjacent culture dishes
Opening's edge form the supporting mouth edge with the support cooperation of the upper bridging arrangement of corresponding upper culture dish.
3. cell according to claim 2 co-cultures unit, it is characterised in that：Upper culture dish in two adjacent culture dishes
On upper bridging arrangement include the overhead gage for circumferentially extending or being distributed along upper culture dish, the upper ventilating structure is arranged on described
On baffle.
4. cell according to claim 3 co-cultures unit, it is characterised in that：Cell co-cultures unit and is additionally provided with head cover,
Cell is co-cultured in unit and is equipped with for head cover overlapped lower bridging arrangement in undermost culture dish, and head cover and cell are total
Interval setting at the opening's edge of the culture dish in top layer in unit is cultivated, is additionally provided on the head cover and/or lower bridging arrangement
The lower ventilating structure being connected to inside and outside head cover.
5. cell according to claim 3 co-cultures unit, it is characterised in that：The downside of the overhead gage is equipped with ring
The limit coordinated along arrangement and with the internal face of lower culture dish and/or the limit of outside wall surface block around the supporting mouth of corresponding lower culture dish
Raised structures.
6. cell according to claim 1 co-cultures unit, it is characterised in that：Upper culture dish is upper in adjacent two culture dish
Bridging arrangement includes the supporter for being sleeved on corresponding lower culture dish outer circumferential, the lower end of the supporter on accordingly lower culture dish
The lower support structure top pressure cooperation being equipped with.
7. cell according to claim 6 co-cultures unit, it is characterised in that：It includes at least that the cell, which co-cultures unit,
Multiple culture dishes of three arrangements stacked on top of one another successively, multiple culture dishes include top culture dish, bottom culture plate and at least one
A intermediate culture dish, at least one intermediate culture dish periphery are installed with medial support structures, which forms in this
Between culture dish as the upper culture dish when upper bridging arrangement, and when forming the intermediate culture dish as the lower culture dish
Lower support structure.
8. cell according to claim 7 co-cultures unit, it is characterised in that：The upper bridging arrangement of the top culture dish
Including overhead gage and the support ring set for being arranged at the opening's edge of overhead gage and overhanging downwards.
9. cell according to claim 7 co-cultures unit, it is characterised in that：The intermediate supports knot of the intermediate culture dish
Structure includes overhead gage and the support ring set that is arranged at the opening's edge of overhead gage and overhangs downwards.
10. cell according to claim 7 co-cultures unit, it is characterised in that：The bottom culture plate periphery is installed with
Lower supporting baffle, the lower supporting baffle form lower support structure of the bottom culture plate as the lower culture dish when, the cell
It includes head cover of the spiral-lock on the culture dish to co-culture unit, and there are ventilative between head cover and the opening's edge of the top culture dish
The lower end support in gap, the head cover is placed in the lower supporting baffle.
11. the cell according to any one of claim 6-10 co-cultures unit, it is characterised in that：The supporter is branch
Pushing out ring set, the lower support structure are equipped with restriction protrusion structure and are limited with the internal face and/or outside wall surface block with support ring set
Position cooperation.
12. the cell according to any one of claim 6-10 co-cultures unit, it is characterised in that：The supporter is edge
The spaced apart multiple support columns of circumferencial direction, multiple support columns cooperatively form interior positioning circumferential surface and/or position circumferential surface outside, described
Lower support structure is equipped with restriction protrusion structure to coordinate with the interior positioning circumferential surface and/or outer positioning circumferential surface block limit.
13. the cell according to claim 3,4,5,8,9 co-cultures unit, it is characterised in that：In two adjacent culture dishes
Upper culture dish overhead gage be equipped with for liquid-transfering gun be inserted into to the connecting hole for descending culture dish adding liquid accordingly, the company
It connects hole and constitutes the upper ventilating structure.
14. cell according to claim 13 co-cultures unit, it is characterised in that：Connecting hole on same overhead gage
Equipped at least two and circumferentially.
15. the cell according to any one of claim 1-10 co-cultures unit, it is characterised in that：Two adjacent culture dishes
In upper culture dish bottom wall be equipped with the interior seepage channel.
16. cell according to claim 15 co-cultures unit, it is characterised in that：Upper culture dish in two adjacent culture dishes
Bottom wall formed by permeable membrane, micropore on permeable membrane constitutes the interior seepage channel.
17. the cell according to any one of claim 1-10 co-cultures unit, it is characterised in that：Each culture dish is interior
Be all provided on the bottom wall of side helpful cell it is adherent help leapfrog structure.
18. the cell according to any one of claim 1-10 co-cultures unit, it is characterised in that：The cell co-cultures
Unit is equipped with the entrance and exit for being connected to outer flow pipe.
19. cell according to claim 18 co-cultures unit, it is characterised in that：The entrance and exit is set at
On undermost culture dish.
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| US20100055790A1 (en) * | 2006-11-14 | 2010-03-04 | Simon Eric M | Cell culture apparatus and associated methods |
| WO2009078637A2 (en) * | 2007-12-18 | 2009-06-25 | Hyo Joo Song | Cell culture container and cell culture system |
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