CN109337813A - Suitable for biological tissue's culture and the system and method for real-time monitoring - Google Patents
Suitable for biological tissue's culture and the system and method for real-time monitoring Download PDFInfo
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- CN109337813A CN109337813A CN201811226234.2A CN201811226234A CN109337813A CN 109337813 A CN109337813 A CN 109337813A CN 201811226234 A CN201811226234 A CN 201811226234A CN 109337813 A CN109337813 A CN 109337813A
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- biological
- organ chip
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y30/00—Apparatus for additive manufacturing; Details thereof or accessories therefor
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- 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
- Y02P10/00—Technologies related to metal processing
- Y02P10/25—Process efficiency
Abstract
The present invention provides a kind of suitable for biological tissue's culture and the system and method for real-time monitoring, is related to the technical field of organizational project, comprising: biological 3D printer, organ chip, connection pedestal, drive system and auxiliary system；Organ chip is connected by connecting pedestal with drive system；Biological 3D printer is for constructing biological 3D printing class loading；Organ chip cultivates the biological 3D printing class loading for placing culture medium and biological 3D printing class loading；Connection pedestal connects drive system for placing organ chip, and drive system is for driving the culture medium to flow in the organ chip, and auxiliary system is for monitoring the histioid state of biological 3D printing.The work such as the system and method are tissue cultures, pathological research and drug screening provide a novel cultivation platform.
Description
Technical field
The present invention relates to field of tissue engineering technology, it is suitable for biological tissue's culture and real-time monitoring more particularly, to a kind of
System and method.
Background technique
Organ chip for the work such as cell or tissue culture, pathological research and drug screening provide one it is novel
Cultivation platform, main characteristics are the reciprocations that can more effectively simulate people's intracorporeal organ.Organ chip is related to three passes
Key elements, respectively living cell tissue/organ element, fluid control elements, detection/sensing element.Fluid control elements provide
Living cell tissue's lifetime bottom and lasting perfusion training mode simulate tumor growth microenvironment；Living cell tissue/organ element
Refer to the component that particular cell types are carried out to specification arrangement on space in 2D or 3D；Detection/sensing element provides monitoring
With Function of Evaluation.
Although compared with 2D situation, the institutional framework of 3D can situation preferably in analogue body, the one of 3D tissue preparation
Cause property, the controllability of dimensional culture, and monitoring property compared with 2D cell line has difficult point in real time, and it is thin how to prepare standardization 3D
Born of the same parents' living tissue, building dimensional culture system and three-dimension monitor scheme are still challenge.It is consistent that biological 3D printing provides a kind of height
Highly controllable organoid method for tissue culture, but in existing organ chip technology, because by biological 3D printing class loading
Directly being printed upon organ chip has inconvenience, so using the organoid of 3D tissue cultures more.
Summary of the invention
In view of this, the purpose of the present invention is to provide the system and the sides that are suitable for biological tissue's culture and real-time monitoring
Method solves organ chip 3D tissue construction, perfusion culture and monitoring and organically combines problem.
In a first aspect, the embodiment of the invention provides it is a kind of suitable for biological tissue culture and real-time monitoring system, packet
It includes: biological 3D printer, organ chip, connection pedestal, drive system and auxiliary system；The organ chip passes through connection pedestal
It is connected with the drive system；
The biology 3D printer is for constructing biological 3D printing class loading；
The organ chip cultivates the biological 3D printing class group for placing culture medium and biological 3D printing class loading
It knits；
The connection pedestal is connected for placing organ chip with drive system；
The drive system is for driving the culture medium to flow in the organ chip；
The auxiliary system is for monitoring the histioid state of biological 3D printing.
With reference to first aspect, the embodiment of the invention provides the first possible embodiments of first aspect, wherein institute
Stating organ chip includes transparent cap, transhipment unit and organ chip body；The transhipment unit is removably embedded in described
The inside of organ chip body, the transparent cap are covered in the organ chip body, and transhipment unit is described for holding
Biological 3D printing class loading.
With reference to first aspect, the embodiment of the invention provides second of possible embodiments of first aspect, wherein institute
State organ chip body further include: hard top layer, miniflow channel layer, transparent underlayer, sensing chip, the miniflow channel layer are arranged in institute
It states between hard top layer and the transparent underlayer, the sensing chip is contacted with the culture medium of organ chip body；
The hard top layer includes at least one culturing room, gas passage, top surface groove, bottom recesses, detection zone；Institute
Bottom recesses are stated for placing the miniflow channel layer；
The miniflow channel layer includes at least one culturing room, fluid channel, driving groove, liquid storage groove, dividing grooves, fence
Shape valve；The fluid channel connects at least one described culturing room, driving groove, liquid storage groove, dividing grooves；It is described
Paliform valve opens dividing grooves partition；
Wherein, the transhipment unit and at least one culturing room in the hard top layer, in the miniflow channel layer extremely
Few culturing room matching.
With reference to first aspect, the embodiment of the invention provides the third possible embodiment of first aspect, wherein device
The shape of official's chip body can be cuboid or hexahedron.
With reference to first aspect, the embodiment of the invention provides the 4th kind of possible embodiments of first aspect, wherein institute
Stating auxiliary system includes: observation system, and/or person's analysis system, and/or person's control system, and/or person's environmental control system；
The observation system is used to observe test organization and culture medium after culture；
The analysis system obtains detection data for being analyzed according to the sensing chip；
The control system is precisely controlled drive system, realizes the control to culture medium flow velocity and flow direction in fluid channel
System；
It is dense that the environmental control system for biological 3D printing class loading growth provides suitable temperature, humidity and carbon dioxide
Degree.
With reference to first aspect, the embodiment of the invention provides the 5th kind of possible embodiments of first aspect, wherein institute
Stating drive system includes display control unit, control device and gas circuit distributor；The control device respectively with the display
Control device, gas circuit distributor are connected；
The display control unit allows the user to multiple works according to viewing display for showing multiple operating modes
Operation mode is selected；
The control device is used for the operating mode selected according to user, controls the gas circuit distributor and works.
With reference to first aspect, the embodiment of the invention provides the 6th kind of possible embodiments of first aspect, wherein institute
The system of stating further includes connection pedestal, and the connection pedestal matches the organ chip and uses, and connection pedestal can pass through connection type
Be combined use with drive system, or connection pedestal and pneumatic system group be combined into an entirety carry out using.
With reference to first aspect, the embodiment of the invention provides the 7th kind of possible embodiments of first aspect, wherein institute
State system further include: more new system, the drive system pass through the more new system and be connected with the organ chip, the driving
System is also used to drive the culture medium in organ chip described in the update system update.
Second aspect, the embodiment of the present invention also provide it is a kind of suitable for biological tissue culture and real-time monitoring method, packet
It includes:
The biological 3D printing class loading of test/test is constructed using biological 3D printer；Wherein, 3D printing class loading conduct
Test organization can directly be printed upon in organ chip culturing room, or be printed upon on transhipment unit and be placed into organ chip culture again
It is indoor；
By receiving the operating mode of user's selection, according to the selected operating mode culture of user in the organ chip
In the test organization；
In incubation, the test organization is detected, detection data is obtained.
In conjunction with second aspect, the embodiment of the invention provides the first possible embodiments of second aspect, wherein institute
The operating mode by receiving user's selection is stated, according to institute of the selected operating mode culture of user in the organ chip
State test organization, comprising:
It is defeated to gas circuit distributor by controlling positive pressure gas source, negative pressure gas source after the operating mode for receiving user's selection
Positive pressure, negative pressure gas flow out, so that the gas passage that positive pressure, negative pressure lead to organ chip transports culture medium by gas circuit distributor
It is dynamic.
In conjunction with second aspect, the embodiment of the invention provides second of possible embodiments of second aspect, wherein logical
The operating mode for receiving user's selection is crossed, according to the examination of the selected operating mode culture of user in the organ chip
After the step of testing tissue, the method also includes:
It is determining that the culture medium in organ chip stores in bubble, according to bubble position, is adjusting work at present mould
Formula.
The embodiment of the present invention brings following the utility model has the advantages that passing through the biological 3D printer of increase in systems, in test,
Biological 3D printer constructs the biological 3D printing class loading for testing/testing, and organ chip places culture medium and biology 3D is beaten
Class loading is printed, the biological 3D printing class loading is cultivated, drive system drives the culture medium to flow in the organ chip,
Auxiliary system monitors biological 3D printing class loading and culture medium, the present invention have test organization with biological 3D printing class loading,
Solve controllability, the consistency of the building of 3D tissue model, and freely construct the bionical histotrophic nutrition/metabolic pathway of complexity can
Row.
Other features and advantages of the present invention will illustrate in the following description, also, partly become from specification
It obtains it is clear that understand through the implementation of the invention.The objectives and other advantages of the invention are in specification, claims
And specifically noted structure is achieved and obtained in attached drawing.
To enable the above objects, features and advantages of the present invention to be clearer and more comprehensible, preferred embodiment is cited below particularly, and cooperate
Appended attached drawing, is described in detail below.
Detailed description of the invention
It, below will be to specific in order to illustrate more clearly of the specific embodiment of the invention or technical solution in the prior art
Embodiment or attached drawing needed to be used in the description of the prior art be briefly described, it should be apparent that, it is described below
Attached drawing is some embodiments of the present invention, for those of ordinary skill in the art, before not making the creative labor
It puts, is also possible to obtain other drawings based on these drawings.
Fig. 1 a is the structure of the system provided by one embodiment of the present invention for being suitable for biological tissue's culture and real-time monitoring
Figure；
Fig. 1 b is the knot for the system for being suitable for biological tissue's culture and real-time monitoring that another embodiment of the present invention provides
Composition；
Fig. 2 is the structure chart of organ chip provided in an embodiment of the present invention；
Fig. 3 a is the positive structure chart of miniflow channel layer provided in an embodiment of the present invention；
Fig. 3 b is the structure chart of the reverse side of miniflow channel layer provided in an embodiment of the present invention；
Fig. 4 is the structure chart of culture medium substitute mode in organ chip provided in an embodiment of the present invention；
Fig. 5 is the structure chart of drive system provided in an embodiment of the present invention；
Fig. 6 is gas circuit distribution map provided in an embodiment of the present invention；
Fig. 7 is the structure chart of connection pedestal provided in an embodiment of the present invention；
Fig. 8 is the structure chart of ventilation mould group provided in an embodiment of the present invention；
Fig. 9 is the flow chart of the method provided in an embodiment of the present invention for being suitable for biological tissue's culture and real-time monitoring.
Specific embodiment
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with attached drawing to the present invention
Technical solution be clearly and completely described, it is clear that described embodiments are some of the embodiments of the present invention, rather than
Whole embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art are not making creative work premise
Under every other embodiment obtained, shall fall within the protection scope of the present invention.
For the organ-tissue adherent growth of existing organ chip in the tissue cultures module of chip, this design can be because of cell
Largely be proliferated, fall off cause cell the death rate increase or block fluid channel.
Existing organ chip mostly uses 2D adherent growth in the tissue cultures module of chip, and this design can be because of cell
It is largely proliferated, falling off causes the death rate of cell to increase or block fluid channel, and the institutional framework of this design and true in vivo
Truth condition differs greatly, the institutional framework of 3D can situation preferably in analogue body, but the consistency of 3D tissue preparation, three-dimensional training
Feeding controllability, and monitoring property compared with 2D cell and tissue structrue module has difficult point in real time, and it is thin how to prepare standardization 3D
Born of the same parents' living tissue, building dimensional culture system and three-dimension monitor scheme are still challenge.
Based on this, it is provided in an embodiment of the present invention it is a kind of suitable for biological tissue culture and real-time monitoring system and side
Method, it is flat that the work such as which is tissue cultures, pathological research and drug screening provide a novel culture
Platform, main characteristics are to provide the class loading organ closer to live tissue, and the interaction for more effectively simulating people's intracorporeal organ is made
With, and real-time continuous monitoring method is provided, and it can be by increasing biological 3D printer in systems, in test, biological 3D is beaten
Printing mechanism builds the biological 3D printing class loading for testing/testing, and organ chip places culture medium and biological 3D printing class loading,
The biological 3D printing class loading is cultivated, drive system drives the culture medium to flow in the organ chip, auxiliary system
It monitors biological 3D printing class loading and culture medium, the present invention has test organization with biological 3D printing class loading, solve thin
The problem of born of the same parents fall off, avoids the blocking of fluid channel, and improves the survival rate of cell, solve 3D tissue model building can
Control property, consistency, and freely construct the feasibility of complicated bionical histotrophic nutrition/metabolic pathway.
To be suitable for biological tissue to one kind disclosed in the embodiment of the present invention first convenient for understanding the present embodiment
The system and method for culture and real-time monitoring describe in detail.
In conjunction with shown in Fig. 1 a, the present invention provides it is a kind of suitable for biological tissue culture and real-time monitoring system, comprising:
Biological 3D printer 1000, organ chip 2000, drive system 3000, auxiliary system 4000 and connection pedestal 5000；Organ core
Piece 2000 is connected by connecting pedestal 5000 with drive system 3000, biological 3D printer 1000, organ chip 2000, driving system
System 3000, auxiliary system 4000 and connection pedestal 5000 can be set on same experimental bench.
Wherein, biological 3D printer 1000 is used to construct the biological 3D printing class loading for testing/testing.Organ chip
2000, for placing culture medium and biological 3D printing class loading, cultivate biological 3D printing class loading.Drive system 3000 is for driving
The culture medium is moved to flow in the organ chip.Auxiliary system 4000 for monitor biological 3D printing class loading and/or
Culture medium.
Specifically, after printing biological 3D printing class loading using biological 3D printer 1000, it is put into organ chip
In 2000, then having biological 3D printing class loading and culture medium in organ chip 2000, drive system driving culture medium exists
It is flowed in organ chip 2000, cultivates biological 3D printing class loading.Auxiliary system can detecte biological 3D printing class loading and/or
Person's culture medium, obtains detection data.
For example, auxiliary system 4000 can detecte cell proliferative conditions in test organization or thin in test organization
Biologic slice result etc. in intracellular growth state or biological 3D printing class loading.
Wherein, the histioid type of biological 3D printing may include but be not limited to: liver, kidney, skin, small intestine, lung,
Tumour and heart.
Organ chip 2000 further includes transparent cap, transhipment unit and organ chip body；It is removably embedding to transport unit
Enter in the inside of the organ chip body, transparent cap is covered in organ chip body, and transhipment unit is described for holding
Biological 3D printing class loading.
As shown in connection with fig. 2, transhipment unit 2100 is embedded in the inside of organ chip body, and organ chip body includes: hard
Matter top layer 2200, miniflow channel layer 2300, transparent underlayer 2400, sensing chip are (due to the training of sensing chip and organ chip body
Base contact is supported, is arranged inside organ chip body, so not marking here), miniflow channel layer 2300 is arranged in hard top layer
Between 2200 and transparent underlayer 2400, sensing chip is contacted with the culture medium of organ chip body, and sensing chip can be set
Between miniflow channel layer 2300 and transparent underlayer 2400.
Wherein, hard top layer 2200 by it is transparent, the material of cytotoxic is made, material can include but is not limited to gather
Polyglass (PS), polycarbonate (PC), cyclenes hydrocarbon type copolymer plastics (COC plastics), polyethylene (PE), polypropylene
(PP)。
Hard top layer 2200 includes at least one culturing room, top surface groove, bottom recesses, detection zone 2210；Bottom surface is recessed
Slot is for placing miniflow channel layer.
At least one culturing room contained in hard top layer can place cell, tissue or biological 3D printing class loading etc. and use
In the trial target of test, can also place containing cell, tissue or the histioid transhipment unit of biological 3D printing；And culturing room
Containing anchor point and positioning eaves, can coincide with the positioning port of transhipment unit.
Top surface groove can distinguish different miniflow channel layer, and cooperate culturing room number carry out using.Detection zone
2210,96 orifice plate enzyme mark strips can be placed, analysis detection are carried out to the culture medium of taking-up, or directly detected in detection zone.
Wherein, transparent cap card slot is for placing transparent cap；
In conjunction with described in Fig. 3 a, Fig. 3 b, miniflow channel layer 2300 includes at least one culturing room 2310, fluid channel 2320, unidirectional
Valve, driving groove 2330, liquid storage groove 2340, dividing grooves 2350, paliform valve 2360；The general of fluid channel 2320 is described at least
One culturing room 2310, check valve, driving groove 2330, liquid storage groove 2340, dividing grooves 2350 connect；Paliform valve
Door 2360 disconnects dividing grooves every 2350.
Soft by dimethyl silicone polymer (PDMS) or comprising dimethyl silicone polymer of miniflow channel layer 2300 and to cell
It grows harmless material to constitute, and its two sides passes through oxygen plasma treatment, irreversible bonding pattern and hard top layer 2200
It is mutually bonded with transparent underlayer 2400.
Fluid channel 2320 connects the culturing room 2310 in miniflow channel layer, width of flow path gradient, dividing grooves 2350 and liquid storage
Groove 2340, and in dividing grooves 2350 by paliform valve 2360 separate or fluid channel 2320 in can be single by addition
The control and containment reflux of flow direction are realized to valve；Diameter≤1mm of fluid channel, the culturing room for miniflow channel layer provide recycle stream
Dynamic fluid nutrient medium provides nutrition for cell, tissue or the histioid growth of biological 3D printing, and metabolic waste is discharged or incites somebody to action
Culture medium containing metabolic waste is discharged into next culturing room；
Optionally, it is properly termed as the width of flow path gradient with the fluid channel of one section of width gradual change in fluid channel 2320,
Width of flow path gradient is greater than in the width close to culturing room one end close to the width of fluid channel one end or the fluid channel and culture
When room is connected by vertical run, then the width of flow path gradient is the fluid channel of one section of Diameter Gradual Change.
Further, the working principle of drive system 3000 are as follows: since drive system 3000 is controlled using the difference of air pressure
What culture medium processed flowed in organ chip 2000, meanwhile, culture medium mainly in hard top layer 2200, miniflow channel layer 2300,
So first introduce the gas circuit of hard top layer 2200.Hard top layer 2200 includes gas passage in the present invention, and gas passage includes gas
Interface tube and gas delivery port, air pipe interface and gas delivery port can be collectively constituted by conduit connection to be led to for a gas
Road, gas delivery port can be connected with the driving groove 2330 of miniflow channel layer 2300；Air pipe interface can pass through pipeline and drive
Dynamic system 3000 is connected, and the gas delivery port on hard top layer is opposite with the position of the driving groove in miniflow channel layer, shape
It answers.In miniflow channel layer 3200, flexible film, which can be used, will drive groove to divide with dividing grooves 2350 or liquid storage groove 2340
It separates；Drive groove 2330 corresponding with the gas delivery port position on hard top layer, shape, and can be by changing gas passage
In air pressure make flexible film deformation occurs, cause dividing grooves 2350 and the volume of liquid storage groove 2340 to change, realize
The control of liquid volume in the opening and closing of paliform valve 2360 and liquid storage groove 2340, to realize culture medium in the organ core
Flowing in piece.
In the case where circulation inside the culture medium in organ chip body, miniflow channel layer can also be had following structure,
In conjunction with shown in Fig. 3 a, fluid channel 3220 can be closed loop runner, and the culture medium in fluid channel is carried out certainly by drive system 3000
It recycles, and realizes the update of culture medium by manually regularly replacing culture medium.The mode manually regularly replaced can be first to take
Former culture medium out, the tool of liquid can be held by, which reusing, is put into new culture medium in organ chip body.
Or the fluid channel can be fabricated in one or more elevation dimensions, i.e., culture medium both can be in the same of runner
It is flowed in a plane, can also be entered by a vertical channel in the runner of another elevation dimension and be continued cycling through.
When carrying out the update of the culture medium in organ chip body using drive system, as shown in connection with fig. 4, drive system
It can be connected by more new system with organ chip 2000, more new system includes liquid storage tank 6100, waste liquid pool 6200, driving system
System is also used to drive the culture medium in organ chip described in the update system update, specifically, drive system 3000 passes through pipe
Road is connected with liquid storage tank 6100, and liquid storage tank 6100 is connected by pipeline with organ chip 2000, waste liquid pool 6200 pass through pipeline and
Organ chip 2000 is connected, and 2200 surface of hard top layer, which can according to need, increases inlet 2220 and liquid outlet 2230.Specifically
For, new culture medium enters inlet 2220 in the case where drive system drives, through pipeline, completes to follow in organ chip 2000
Organ chip 2000 is left from liquid outlet 2230 by pipeline after ring.
Wherein, the transhipment unit and at least one culturing room in the hard top layer, in the miniflow channel layer extremely
Few culturing room matching.
Wherein, hard top layer 2200, miniflow channel layer 2300, transparent underlayer 2400 is by oxygen plasma treatment, with irreversible
Bonding pattern is bonded, and in bonding process hard top layer 2200 culturing room, be with the culture of miniflow channel layer 2300
2310 Corresponding matching of room, and sensing chip is fixed on miniflow in bonding process by miniflow channel layer 2300 and transparent underlayer 2400
Between channel layer 2300 and transparent underlayer 2400, and it can be in contact with the culture medium in the fluid channel 2320 of miniflow channel layer 2300, device
Official's chip body carry out as a whole using.
Transparent underlayer 2400 can be made of the material of the glass of clear hard or other clear hards, with miniflow channel layer 2300
By being irreversibly bonded, substrate is provided for miniflow channel layer 2300, and include sensing chip.
Sensing chip is the surface-functionalized biochip for having biomarker, is placed in fluid channel, hard top layer or micro-
It the top or bottom end of culturing room and is contacted with culture medium in flow channel layer；According to sensing needed for selected sensing principle and sensing chip
Condition, organ chip body is equipped with the channel that organ chip is passed in and out for sensed condition, and makes auxiliary system and sensing chip phase
Connection；The sensed condition includes but is not limited to light, electric current, voltage, magnetic field, and each side's region feature of organ chip is real-time
Ground detection and record.
In addition, biological 3D printer is in addition to can directly be printed upon in the culturing room on hard top layer；Or by the life
Object 3D printing class loading is printed upon in transhipment unit, is subsequently placed in the culturing room in organ chip body.
As shown in connection with fig. 5, drive system 3000 includes display control unit 3100, control device 3200, gas circuit distribution dress
Set 3300；
Wherein, display control unit 3100 may include touch screen 3130, indicator light 3110, switch 3120, fill with control
It sets and carries out information connection, for display control unit for showing multiple operating modes, user can be by viewing touch screen 3130
Operating mode, carry out the operating mode wanted of selection user.Indicator light 3110 can be used to indicate that touch screen can be made
With situation is obtained, switch 3120 can be used for starting display control unit.
The control device is used for the operating mode selected according to user, and control gas circuit distributor works.Specifically
For, in conjunction with shown in Fig. 5, control device 3200 is defeated to gas circuit distributor 3300 by control positive pressure gas source, negative pressure gas source
Positive pressure, negative pressure gas flow out, so that positive pressure, negative pressure, are led to the gas passage of organ chip body by gas circuit distributor 3300.Gas
Road distributor 3300 is connected by fast plug and tracheae with organ chip body, is at least connected with an organ chip body
It connects；The gas circuit distributor controls positive pressure, the negative pressure of the air-flow of output to organ chip body, by four steps
The circulation action of composition makes culture medium according to flowing clockwise or counterclockwise, and when culture medium clockwise flow, the circulation is
Right-hand circular, when culture medium counterclockwise flow, the circulation is left-hand circular.Wherein, the circulation of four steps composition,
It is to be completed by three driving groove positive pressures of control or negative pressure output；
There are the first dividing grooves, liquid storage groove and the second dividing grooves below the corresponding flexible film of three driving grooves；
As shown in connection with fig. 6, when work, three corresponding air pressures of driving groove are respectively as follows: step 1, export positive pressure, negative pressure, negative pressure；Step
2 output negative pressure, positive pressure, negative pressure；Step 3 exports negative pressure, positive pressure, positive pressure；Step 4 exports negative pressure, negative pressure, positive pressure；Complete culture
Base is from the first dividing grooves to the driving in the second dividing grooves direction.
Wherein, operating mode selection includes liquid feeding mode, alternate mode, circulation pattern, manual mode and step mode；
Liquid feeding mode refers to that all gas passages are provided which negative pressure, is raised all flexible films of miniflow channel layer, makes
All paliform valve openings, liquid storage depression volume increase, and culture medium is added for organ chip.
Alternate mode refers to can set gas circuit distributor by parameter setting, what realization recycled clockwise or counterclockwise
Alternately.In actually control, the control device exports positive pressure and/or negative pressure and gives gas circuit distributor, gas circuit distributor
Carry out gas circuit output so that culture medium according to clockwise flow for a period of time, for a period of time according still further to counterclockwise flow, and recycle
Alternately.
Circulation pattern is to set the lasting progress of gas circuit distributor by parameter setting to recycle clockwise or counterclockwise.?
When practical control, the control device exports positive pressure and/or negative pressure and gives gas circuit distributor, and gas circuit distributor carries out gas circuit
Output, so that culture medium is according to clockwise flow or counterclockwise flow.
Manual mode refers to the positive pressure output and negative pressure output that all gas channels are manually controlled by display control unit.
Step mode refer to by display control unit by gas channels status be adjusted to it is set counterclockwise or
The previous step or next step of right-hand circular.For example, when currently being flowed for clockwise direction, in step mode
Under, it can pass through four times operations by operating the step of making gas channels status enter next right-hand circular and complete
Right-hand circular；Or when can flow in the clockwise direction, gas channels status is set to enter upper one by operation
The step of a right-hand circular, completes a left-hand circular by four operations.
Wherein, auxiliary system 4000 includes: observation system and/or analysis system.Observation system is used for observation experiment group
It knits and culture medium；Analysis system obtains detection data for being analyzed according to sensing chip.
In addition, auxiliary system further includes control system, environmental control system.
Wherein, control system is precisely controlled drive system 3000, realizes to culture medium flow velocity and stream in fluid channel
To control；Swap time by controlling media flow realizes organ chip body culture indoor biological 3D printing class loading
It is reacted with substance in culture solution.
Wherein, observation system is completed using the high grade of transparency of organ chip to the biological 3D printing class group cultivated in culturing room
Knit the function of directly observing with the culture medium in fluid channel.
Wherein, observation system can choose but be not limited only to following methods: optical microscopy, fluorescence microscope, difference are aobvious
Micro mirror, optical coherence tomography system (OCT)；
Environmental control system provides suitable temperature, humidity and gas concentration lwevel for tissue growth.Wherein, the present invention mentions
One incubator is set around the organ chip of confession, and environmental control system is mainly that suitable tissue cultures are provided in incubator
Temperature, humidity and gas concentration lwevel etc. environmental factor.
Furthermore it is possible to by being connected into the sensing core for capableing of the indexs such as real-time monitoring pH, temperature, flow velocity in fluid channel
The information such as pH, temperature, flow velocity are sent to environmental control system by piece, and environmental control system can be according to collected by pH, temperature
The information such as degree, flow velocity are monitored, and when discovery overflows situation, can remind user by alarm.
Analysis system can analyze the signal obtained on sensing chip, and provide one kind indirectly by analyzing result
Real-time online monitoring is carried out to the biological 3D printing class loading cultivated on organ chip；
Wherein, when the signal that sensing chip is exported is optical signal, which need to have the wavelength to light, function
One of parameters such as rate, energy, incident angle, pulsewidth, repetition, degree of polarization or a variety of detectabilities；
When the signal that sensing chip is exported is electric signal, which need to have the voltage, electric current, electricity to circuit
One or more detectabilities of the parameters such as lotus amount.
In conjunction with shown in Fig. 1, connection pedestal 5000 matches organ chip 2000 and uses.The connection pedestal can pass through connection
Mode and the drive system are combined uses, or connection pedestal and drive system group be combined into an entirety carry out using.Again
As shown in connection with fig. 1, drive system 3000 with connect pedestal by pipeline 3500 be connected.
As shown in connection with fig. 7, pedestal 5000, including base cover 5010, fixed bayonet 5020, card slot 5030, ventilation are connected
Mould group 5040, air guide connector 5042, spring stack, quick coupling 5044, gas-guide tube 5045, sliding group 5050, is seen ventilation sliding block 5041
Examine window, shaft 5070；
Wherein, as shown in connection with fig. 8, the gas-guide tube for mould group 5040 of ventilating is connected in quick coupling 5044, and the air guide connects
First 5042 and quick coupling 5044 be connected to ventilation sliding block 5041 two sides, by ventilation sliding block inside gas-guide tube mutually interconnect
It is logical；The ventilation mould group 5040 is connected by sliding group 5050 with card slot 5030, and sliding group 5050, ventilation mould group 5040 can opposite cards
Slot sliding.
Base cover 5010 is connected with card slot 5030 by shaft 5070, and base cover 5010 can push away during covering
Dynamic ventilation 5040 forward slip of mould group；Base cover is opened, is opened to 90 degree or more, by the air pipe interface of organ chip towards air guide
The direction of connector 5042 is put into the card slot 5030 of connection pedestal, is pressed downward base cover 5010 with hand, base cover 5010
Ventilation 5040 forward slip of mould group is pushed during covering, air guide connector 5042 is inserted into the air pipe interface of organ chip body
In, it is sealed by the O-ring seal on air guide connector 5042.After covering base cover 5010, fixed bayonet 5020 is set right, Gu
Surely base cover 5010 is lived, prevents from loosening.
Other than the form of Fig. 1 a, the present invention also provides a kind of organ chips of shape described in combination Fig. 1 b, can make
It include transparent cap 2500 on organ chip 2000 for hexahedron, organ chip 2000 can pass through connection pedestal 5000 and drive
Dynamic 3000 groups of system is combined into an entirety, and drive system 3000 is connect with display control unit 3100, passes through display control unit
3100 carry out control operation.
As shown in connection with fig. 9, the present invention also provides it is a kind of suitable for biological tissue culture and real-time monitoring method, packet
It includes:
S110: the biological 3D printing class loading of test/test is constructed using biological 3D printer；Wherein, it is directly printed upon
In organ chip culturing room, or it is printed upon on transhipment unit and is placed into organ chip culturing room again；
S120: the operating mode by receiving user's selection, according to the selected operating mode culture experiment tissue of user.
Specifically, after the operating mode for receiving user's selection, by controlling positive pressure gas source, negative pressure gas source, to gas circuit
Distributor exports positive pressure, negative pressure gas flow, so that gas circuit distributor makes the gas passage that positive pressure, negative pressure lead to organ chip
Culture medium movement.
Specifically, after the liquid feeding mode for receiving user's selection, the culture medium controlled in liquid storage tank or culturing room enters
In fluid channel；
Receive user selection alternate mode after, by clockwise or counterclockwise alternately in a manner of, control organ core
Media flow in piece main body；
After the circulation pattern for receiving user's selection, in clockwise manner or counter-clockwise, organ chip body is controlled
In culture medium flowed；
After the step mode for receiving user's selection, primary flowing before the loop direction that current media flows is adjusted to
Direction or a rear flow direction.
S130: in incubation, the test organization is detected, detection data is obtained.
It is introduced below it is several utilize it is provided by the invention suitable for biological tissue culture and real-time monitoring system and method
Practical application example.
Example 1
Step 1. takes a transhipment unit 2100, and the biological 3D with liver cell is printed in it with biological 3D printer 1000
Class loading is printed, and is put into culture 20 days in culture medium (+1% mycillin of+10% fetal calf serum of DMEM culture medium), it is during which every
Two days one subcultures of replacement；
Step 2. separately takes a transhipment unit 2100, is printed in it with biological 3D printer 1000 with tumour cell
Biological 3D printing class loading, and culture 20 days in culture medium (+1% mycillin of+10% fetal calf serum of DMEM culture medium) are put into,
One subculture of every two days of period replacement；
Step 3. will contain liver cell biology 3D printing class loading and be put into containing tumour cell biology 3D printing class loading
It transports in unit 2100, and transhipment unit 2100 is installed to two different culturing room on organ chip 2000；
Organ chip 2000 is put into connection pedestal 5000 by step 4., closes base cover 5010 with by air guide connector
5042 insertion air pipe interfaces；
The pipeline 3500 of fast joint 3400 in drive system 3000 is connected by step 5., makes drive system 3000 and connects
Pedestal 5000 is connect to be connected；
Step 6. turns on the switch 3120, and indicator light 3110 is waited to light, and enters drive system 3000 using touch screen 3130
It carries out in operating mode selection, selects liquid feeding mode, then add culture medium into transhipment unit 2100 with syringe or liquid-transfering gun,
It allows culture medium to flow into culturing room, terminate liquid feeding mode after culture medium is full of entire fluid channel 2320 and returns to operating mode selection
Interface；
Step 7. opens base cover 5010 and closes pedestal again after transparent cap is placed on organ chip 2000
Lid 5010 is air guide connector 5042 to be inserted on air pipe interface；
Step 8. selects circulation pattern in drive system 3000, and be arranged loop direction be it is clockwise, cycle frequency is
0.5Hz, positive pressure output valve are 150kPa, and negative pressure exports 65kPa, later on circulation pattern；
Step 9. environmental control system provides isoperibol and control gas concentration lwevel in this specific embodiment,
Connection pedestal 5000 containing organ chip 2000 is put into incubator, the drive system 3000 outside pipeline and incubator is passed through
Connection is kept, the environment of incubator is set as 37 DEG C, gas concentration lwevel 5%；
Step 10. is analyzed by the biological information that analysis system spreads out of sensing chip to monitor in fluid channel 2320
Medium component simultaneously infers cell growth condition；
After step 11.6 hour, opens connection pedestal 5000 of the culture chamber door taking-up containing organ chip 2000 and be placed on down
It sets under optical microscopy, cellular morphology on culture indoor biological 3D printing class loading is observed using watch window, is opened
Base cover 5010, removes transparent cap and culture medium samples in culturing room with liquid-transfering gun, and carries out cell Proliferation to sample
Detection；
The histioid culturing room of the 3D printing of biology containing liver cell is added in institute's testing drug by step 12.；
Transparent cap is placed on organ chip 2000 by step 13., then closes base cover 5010 for air guide connector
Connection pedestal 5000 containing organ chip 2000 is simultaneously put into incubator by 5042 insertion air pipe interfaces；
Step 14. repeats step 10-13 three times；
After step 15.6 hour, step 10-11 is repeated, is then removed culture medium in all culturing room with liquid-transfering gun, and
New culture medium is added；
After step 16. repetition step 14-15 is primary, it is primary to repeat step 14；
After step 17.6 hour, stop circulation pattern in drive system 3000, by the connection containing organ chip 2000
Pedestal 5000 takes out from incubator, opens base cover 5010, removes transparent cap, takes out all culture mediums in culturing room and makees
For sample, celliferous biological 3D printing class loading taking-up is further tested；
Step 18. observes and records the record of sensing obtained in experimentation, Cell proliferation results, cell growth state
And the biological histioid biologic slice result of 3D printing summarizes and analyzes liver cell on the biological 3D printing class loading of acquisition and swells
The growth conditions of oncocyte during the experiment, the liver cell if growth of the addition inhibition tumour cell of drug does not have an impact
Growth, then the drug is screened by this hepatotoxicity wind agitation.
Example 2
2 transhipment units 2100 are respectively put into 2 culturing room on organ chip 2000 by step 1.；
Step 2. enters printing syringe needle in culturing room by adjusting the printing head of biological 3D printer 1000；
Step 3. is respectively by the biological 3D printing class loading with liver cell and the biological 3D printing class loading with tumour cell
It is printed upon on the transhipment unit 2100 in 2 culturing room；
Step 4. is added culture medium (+1% mycillin of+10% fetal calf serum of DMEM culture medium) into culturing room, is executed
The operation of step 4-7 in example 1；
Step 5. is cultivated cell 20 days, carries out culture medium replacement by step 6 in embodiment 1 within during which every two days；
Step 6. executes step 8-17 in example 1；
Step 7. observes and records the record of sensing obtained in experimentation, Cell proliferation results, cell growth state
And the biological histioid biologic slice result of 3D printing summarizes and analyzes liver cell on the biological 3D printing class loading of acquisition and swells
The growth conditions of oncocyte during the experiment, the liver cell if growth of the addition inhibition tumour cell of drug does not have an impact
Growth, then the drug is screened by this hepatotoxicity wind agitation.
Example 3
Step 1. executes step 1-10 in example 1；
Step 2. is when marker concentrations are insufficient or sensing chip reaction speed deficiency causes culture medium to flow through, the biography of acquisition
It is too low to feel signal, stops circulation pattern in drive system 3000, and switch to alternate mode, after 3 right-hand circulars are set
It pauses 5 seconds and carries out 2 left-hand circulars, cycle frequency 0.5Hz, positive pressure output valve is 150kPa, and negative pressure output valve is 65kPa,
Later on alternate mode；
Step 3. executes step 9-17 in example 1, wherein " stops circulation pattern in drive system 3000 " in step 17
It is changed to " stopping alternate mode in drive system 3000 "；
Step 4. observes and records the record of sensing obtained in experimentation, Cell proliferation results, cell growth state
And the biological histioid biologic slice result of 3D printing summarizes and analyzes liver cell on the biological 3D printing class loading of acquisition and swells
The growth conditions of oncocyte during the experiment, the liver cell if growth of the addition inhibition tumour cell of drug does not have an impact
Growth, then the drug is screened by this hepatotoxicity wind agitation.
Example 4
Step 1. executes step 1-8 in example 1；
Step 2. stops circulation pattern in drive system 3000 and selects to walk when finding has bubble in fluid channel 2320
Progressive die formula, loop direction, which is adjusted to bubble in miniflow channel layer 2300, can enter culturing room without paliform valve 2360
Direction manually controls step mode and slowly bubble is passed through in culturing room through discharge at the top of culture medium liquid level；
Step 3. exits step mode in drive system 3000, selects circulation pattern and cycle frequency is arranged as 0.5Hz,
Positive pressure output valve is 150kPa, and negative pressure exports 65kPa, later on circulation pattern；
Step 4. executes step 9-17 in example 1；
Step 5. observes and records the record of sensing obtained in experimentation, Cell proliferation results, cell growth state
And the biological histioid biologic slice result of 3D printing summarizes and analyzes liver cell on the biological 3D printing class loading of acquisition and swells
The growth conditions of oncocyte during the experiment, the liver cell if growth of the addition inhibition tumour cell of drug does not have an impact
Growth, then the drug is screened by this hepatotoxicity wind agitation.
Example 5
Step 1. executes step 1-8 in example 1
Step 2. stops in drive system 3000 when finding has bubble in dividing grooves 3250 or liquid storage groove 3240
Circulation pattern simultaneously selects manual mode；
The positive pressure and negative pressure that step 3. manually controls drive mode output are to control dividing grooves 2350 and liquid storage groove
Bubble is squeezed into fluid channel 2320 from dividing grooves 2350 or liquid storage groove 2340 by 2340 volume；
Step 4. stops circulation pattern in drive system 3000 and selects step mode, and loop direction is adjusted to miniflow
Bubble can enter the direction of culturing room without paliform valve 2360 in channel layer 2300, manually control step mode and slowly will
Bubble is passed through in culturing room and the discharge at the top of culture medium liquid level；
Step 5. exits step mode in drive system 3000, selects circulation pattern and cycle frequency is arranged as 0.5Hz,
Positive pressure output valve is 150kPa, and negative pressure exports 65kPa, later on circulation pattern；
Step 6. executes step 9-17 in example 1；
Step 7. observes and records the record of sensing obtained in experimentation, Cell proliferation results, cell growth state
And the biological histioid biologic slice result of 3D printing summarizes and analyzes liver cell on the biological 3D printing class loading of acquisition and swells
The growth conditions of oncocyte during the experiment, the liver cell if growth of the addition inhibition tumour cell of drug does not have an impact
Growth, then the drug is screened by this hepatotoxicity wind agitation.
Example 6
Step 1. executes step 1-14 in example 1；
After step 2.6 hour, opens connection pedestal 5000 of the culture chamber door taking-up containing organ chip 2000 and be placed on down
It sets under optical microscopy, cellular morphology on culture indoor biological 3D printing class loading is observed using watch window, is opened
Base cover 5010, removes transparent cap and culture medium samples in culturing room with liquid-transfering gun, and detection zone is added in sample, or
After being caught in enzyme mark strip in the detection area, enzyme mark strip is added in sample, and add marker；
Organ chip 2000 is put into microplate reader by step 2., carries out analysis test；
Step 3. takes out organ chip 2000 from microplate reader, and executes step 15-17 in example 1；
Step 4. observes and records the record of sensing obtained in experimentation, Cell proliferation results, cell growth state
And the biological histioid biologic slice result of 3D printing summarizes and analyzes liver cell on the biological 3D printing class loading of acquisition and swells
The growth conditions of oncocyte during the experiment, the liver cell if growth of the addition inhibition tumour cell of drug does not have an impact
Growth, then the drug is screened by this hepatotoxicity wind agitation.
Example 7
Step 1. executes step 1-3 in example 1
Step 2. opens drive system 3000, the culture medium in liquid storage tank 6100 is pumped into pipeline, by pipeline and inlet
2220 and liquid outlet 2230 connect, make culture medium by inlet 2220 enter fluid channel 2320, culture medium be full of all cultures
Behind room, after leaving fluid channel 2320 by liquid outlet 2230, waste liquid pool 6200 is flowed by pipeline；
Step 3., which adjusts drive system 3000, makes flow velocity be fixed on 20 microlitres/second；
Step 4. will containing there are two transhipment unit 2100 and being stamped the organ chip 2000 of transparent cap and be put into incubator, and
And make culturing room locating for the transhipment unit 2100 containing tumour cell than training locating for the transhipment unit 2100 containing liver cell
Support room in the runner of culture medium be in more upstream position, 37 DEG C of set temperature, gas concentration lwevel 5%；
Step 5. is analyzed by the biological information that analysis system spreads out of sensing chip to monitor in fluid channel 2320
Medium component simultaneously infers cell growth condition；
Organ chip 2000 is taken out from incubator after step 6.6 hour, waste liquid pool 6200 is connect disconnection with pipeline,
From taking the culture medium flowed out in liquid outlet 2230 to be further analyzed in pipeline；
The culture medium of the testing drug containing institute is added in step 7. in liquid storage tank 6100；
Step 8. repeats step 5-6；
After step 9. experiment starts 72 hours, is stopping drive system 3000, organ chip 2000 is being taken from incubator
Out；
Step 10. observes and records the record of sensing obtained in experimentation, Cell proliferation results, cell growth state
And the biological histioid biologic slice result of 3D printing summarizes and analyzes liver cell on the biological 3D printing class loading of acquisition and swells
The addition of the growth conditions of oncocyte during the experiment, drug does not have an impact the growth of liver and tumour cell, obtains
The conclusion drug is invalid to tumour out or can not directly effectively.
Example 8
Step 1. executes step 1-3 in example 1
Step 2. opens drive system 3000, the culture medium in liquid storage tank 6100 is pumped into pipeline, by pipeline and inlet
2220 and liquid outlet 2230 connect, make culture medium by inlet 2220 enter fluid channel 2320, culture medium be full of all cultures
Behind room, fluid channel 2320 is left by liquid outlet 2230, waste liquid pool 6200 is flowed by pipeline；
Step 3., which adjusts drive system 3000, makes flow velocity be fixed on 20 microlitres/second；
Step 4. will containing there are two transhipment unit 2100 and being stamped the organ chip body of transparent cap and be put into incubator, and
And make culturing room locating for the transhipment unit 2100 containing tumour cell than training locating for the transhipment unit 2100 containing liver cell
Support room in the runner of culture medium be in further downstream position, 37 DEG C of set temperature, gas concentration lwevel 5%；
Step 5. is analyzed by the biological information that analysis system spreads out of sensing chip to monitor in fluid channel 2320
Medium component simultaneously infers cell growth condition；
Organ chip 2000 is taken out from incubator after step 6.6 hour, waste liquid pool 6200 is connect disconnection with pipeline,
From taking the culture medium flowed out in liquid outlet 2230 to be further analyzed in pipeline；
The culture medium of the testing drug containing institute is added in step 7. in liquid storage tank 6100；
Step 8. repeats step 5-6；
After step 9. experiment starts 72 hours, is stopping drive system 3000, organ chip 2000 is being taken from incubator
Out；
Step 10. observes and records the record of sensing obtained in experimentation, Cell proliferation results, cell growth state
And the biological histioid biologic slice result of 3D printing summarizes and analyzes liver cell on the biological 3D printing class loading of acquisition and swells
The growth conditions of oncocyte during the experiment.The addition of drug inhibits the growth of tumour cell and the life without influencing liver cell
It grows, in conjunction with the embodiments 7 resulting experimental result, it was demonstrated that the drug is needed by that can generate drug effect to tumour after liver metabolism,
And there is no hepatotoxicity wind agitation.
Example 9
Step 1. takes a transhipment unit 2100, and the biological 3D with liver cell is printed in it with biological 3D printer 1000
Class loading is printed, and is put into culture 20 days in culture medium (+1% mycillin of+10% fetal calf serum of DMEM culture medium), it is during which every
Two days one subcultures of replacement；
Step 2. takes another to transport unit 2100, and the biology with nephrocyte is printed in it with biological 3D printer 1000
3D printing class loading, and culture 20 days in culture medium (+1% mycillin of+10% fetal calf serum of DMEM culture medium) are put into, during which
Every two days one subcultures of replacement；
Step 3. will contain liver cell biology 3D printing class loading and contain the histioid transhipment of nephrocyte biology 3D printing
Unit 2100 is respectively put into 2 culturing room in organ chip body；
Step 4. executes step 4-9 in example 1；
The biological information and concentration of glucose that step 5. spread out of sensing chip by analysis system are analyzed with prison
It controls medium component in fluid channel 2320 and infers cell growth condition；
After step 6.6 hour, opens connection pedestal 5000 of the culture chamber door taking-up containing organ chip 2000 and be placed on down
It sets under optical microscopy, cellular morphology on culture indoor biological 3D printing class loading is observed using peep hole, opens bottom
Flap 5010, removes transparent cap and culture medium samples in culturing room with liquid-transfering gun, and carries out cell Proliferation inspection to sample
It surveys；
After step 7. is removed culture medium in all culturing room with liquid-transfering gun, the culture of the glucose containing 15mmol/L is added
Base；
Transparent cap is placed on organ chip 2000 by step 8., then closes base cover 5010 for air guide connector
Connection pedestal 5000 containing organ chip 2000 is simultaneously put into incubator by 5042 insertion air pipe interfaces；
Step 9. repeats step 5-8, until finding that lesion occurs for liver cell or nephrocyte by observation or sensing data；
Step 10. stops circulation pattern in drive system 3000, by the connection pedestal 5000 containing organ chip 2000
It is taken out from incubator, opens base cover 5010, remove transparent cap, take out in culturing room all culture mediums as sample,
Celliferous biological 3D printing class loading taking-up is further tested；
Step 11. observes and records the record of sensing obtained in experimentation, Cell proliferation results, cell growth state
And the biological histioid biologic slice result of 3D printing summarizes and analyzes liver cell and kidney on the biological 3D printing class loading of acquisition
The growth conditions of cell during the experiment, and diabetes are summarized to the induced factor of liver, renal lesions.
Embodiment 10
Step 1. takes a transhipment unit 2100, and the biological 3D with liver cell is printed in it with biological 3D printer 1000
Class loading is printed, and is put into culture 20 days in culture medium (+1% mycillin of+10% fetal calf serum of DMEM culture medium), it is during which every
Two days one subcultures of replacement；
Step 2. separately takes a transhipment unit 2100, prints the biology with tumour cell in it with biological 3D printer
3D printing class loading, and culture 20 days in culture medium (+1% mycillin of+10% fetal calf serum of DMEM culture medium) are put into, during which
Every two days one subcultures of replacement；
Step 3. will contain liver cell biology 3D printing class loading and contain histioid turn of tumour cell biology 3D printing
Transportation unit 2100, two different culturing room being respectively put on organ chip, and with transparent cap organ chip is carried out close
Envelope；
Organ chip is placed on connection pedestal 5000 by step 4., and it is integrated with drive system 3000 to connect pedestal 5000
In the same machine；
Drive system 3000 is connected by step 5. with touch screen 3130, and drive system is arranged by touch screen 3130
3000, circulation pattern is selected, flow velocity is set as 30uL/min, disconnects drive system 3000 and touch screen 3130 after setting completed
Connection；
Step 9. environmental control system can provide isoperibol, control gas concentration lwevel in this specific embodiment
And chamber door has the incubator of string holes, and the drive system 3000 for being connected with organ chip and connection pedestal 5000 is put into incubator
In, the environment of incubator is set as 37 DEG C, gas concentration lwevel 5%；
Step 10. is analyzed by the biological information that analysis system spreads out of sensing chip to monitor in fluid channel 2320
Medium component simultaneously infers cell growth condition；
After step 11.6 hour, opens the taking-up of incubator goalkeeper organ chip 2000 and is placed under inverted light microscope,
Cellular morphology on culture indoor biological 3D printing class loading is observed, removes transparent cap and with liquid-transfering gun in culturing room
Culture medium sampling, and cell Proliferation detection is carried out to sample；
The histioid culturing room 3110 of the 3D printing of biology containing liver cell is added in institute's testing drug by step 12.,
Transparent cap is placed on organ chip by step 13., then turns on incubator, and organ chip is connected to connection
On pedestal 5000, incubator is closed；
Step 14. repeats step 10-13 three times；
After step 15.6 hour, step 10-11 is repeated, is then removed culture medium in all culturing room with liquid-transfering gun, and
New culture medium is added；
After step 16. repetition step 14-15 is primary, it is primary to repeat step 14；
After step 17.6 hour, the drive system 3000 for being connected with organ chip and connection pedestal 5000 is taken from incubator
Out, transparent cap is removed, all culture mediums in culturing room is taken out and takes out celliferous biological 3D printing class loading as sample
It is further tested, touch screen 3130 is connected with drive system 3000, stop circulation pattern；
Step 18. observes and records the record of sensing obtained in experimentation, Cell proliferation results, cell growth state
And the biological histioid biologic slice result of 3D printing summarizes and analyzes liver cell on the biological 3D printing class loading of acquisition and swells
The growth conditions of oncocyte during the experiment, the liver cell if growth of the addition inhibition tumour cell of drug does not have an impact
Growth, then the drug is screened by this hepatotoxicity wind agitation.
The present invention combines organ chip technology with biological 3D printing technique, adherent to improve existing organ chip cell
The training mode of growth.Specifically the cell for cultivating organ needed for organ chip is mixed in biological 3D printing ink, is printed
Biological 3D printing class loading with cell is cultivated.Such a process increases the organization survival times, and cell detachment is avoided to block
Organ chip runner, and a kind of cultural method of more approximate tumor growth environment is provided, the model for constructing organ chip
More simulate human body.
Meanwhile, it is capable to by can effectively assess biological 3D printing class group to the direct observation of organ chip and on-line monitoring
The growth conditions and growing environment knitted, thus provide more to biological 3D printing histioid growth course analysis data or
Biological 3D printing class loading growth executes emergency in time when something goes wrong and puts on record to reduce loss.In addition, organ chip can be straight
It scoops out for detecting instruments such as microplate reader, convenient for operation.
Thirdly, the present invention can also real-time online the histioid growth conditions of detection biology 3D printing and metabolin contain
Amount, at the same realize organ chip tissue cultures, drug screening, pathological study operation automation, reduce human factor influence.
4th point, transhipment unit proposed by the invention provides the hand of transhipment between 3D printer and organ chip
Section, avoids the technical difficulty directly printed in organ chip.The connection pedestal proposed can carry out the fast insert-pull of tracheae,
Reduce the troublesome operation of pneumatic type organ chip plug tracheae.
Unless specifically stated otherwise, the opposite step of the component and step that otherwise illustrate in these embodiments, digital table
It is not limit the scope of the invention up to formula and numerical value.
In all examples being illustrated and described herein, any occurrence should be construed as merely illustratively, without
It is as limitation, therefore, other examples of exemplary embodiment can have different values.
It should also be noted that similar label and letter indicate similar terms in following attached drawing, therefore, once a certain Xiang Yi
It is defined in a attached drawing, does not then need that it is further defined and explained in subsequent attached drawing.
In addition, in the description of the embodiment of the present invention unless specifically defined or limited otherwise, term " installation ", " phase
Even ", " connection " shall be understood in a broad sense, for example, it may be being fixedly connected, may be a detachable connection, or be integrally connected；It can
To be mechanical connection, it is also possible to be electrically connected；It can be directly connected, can also can be indirectly connected through an intermediary
Connection inside two elements.For the ordinary skill in the art, above-mentioned term can be understood at this with concrete condition
Concrete meaning in invention.
In the description of the present invention, it should be noted that term " center ", "upper", "lower", "left", "right", "vertical",
The orientation or positional relationship of the instructions such as "horizontal", "inner", "outside" be based on the orientation or positional relationship shown in the drawings, merely to
Convenient for description the present invention and simplify description, rather than the device or element of indication or suggestion meaning must have a particular orientation,
It is constructed and operated in a specific orientation, therefore is not considered as limiting the invention.In addition, term " first ", " second ",
" third " is used for descriptive purposes only and cannot be understood as indicating or suggesting relative importance.
Finally, it should be noted that embodiment described above, only a specific embodiment of the invention, to illustrate the present invention
Technical solution, rather than its limitations, scope of protection of the present invention is not limited thereto, although with reference to the foregoing embodiments to this hair
It is bright to be described in detail, those skilled in the art should understand that: anyone skilled in the art
In the technical scope disclosed by the present invention, it can still modify to technical solution documented by previous embodiment or can be light
It is readily conceivable that variation or equivalent replacement of some of the technical features；And these modifications, variation or replacement, do not make
The essence of corresponding technical solution is detached from the spirit and scope of technical solution of the embodiment of the present invention, should all cover in protection of the invention
Within the scope of.Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (11)
1. it is a kind of suitable for biological tissue culture and real-time monitoring system characterized by comprising biological 3D printer, device
Official's chip, connection pedestal, drive system and auxiliary system；The organ chip passes through the connection pedestal and the drive system
It is connected；
The biology 3D printer is for constructing biological 3D printing class loading；
The organ chip cultivates the biological 3D printing class loading for placing culture medium and biological 3D printing class loading；
The connection pedestal is connected for placing organ chip with drive system；
The drive system is for driving the culture medium to flow in the organ chip；
The auxiliary system is for monitoring the histioid state of biological 3D printing.
2. system according to claim 1, which is characterized in that the organ chip include transparent cap, transhipment unit and
Organ chip body；The transhipment unit is removably embedded in the inside of the organ chip body, and the transparent cap covers
It covers in the organ chip body, transhipment unit is for holding the biological 3D printing class loading.
3. system according to claim 2, which is characterized in that the organ chip body further include: hard top layer, miniflow
Channel layer, transparent underlayer, sensing chip, the miniflow channel layer is arranged between the hard top layer and the transparent underlayer, described
Sensing chip is contacted with the culture medium of organ chip body；
The hard top layer includes at least one culturing room, gas passage, top surface groove, bottom recesses, detection zone；The bottom
Face groove is for placing the miniflow channel layer；
The miniflow channel layer includes at least one culturing room, fluid channel, driving groove, liquid storage groove, dividing grooves, paliform valve
Door；The fluid channel connects at least one described culturing room, driving groove, liquid storage groove, dividing grooves；The fence
Shape valve opens dividing grooves partition；
Wherein, described to transport at least one culturing room in unit and the hard top layer, at least one in the miniflow channel layer
A culturing room's matching.
4. system according to claim 2, which is characterized in that the shape of the organ chip body is cuboid or six faces
Body.
5. system according to claim 1, which is characterized in that the auxiliary system includes: observation system, and/or person point
Analysis system, and/or person's control system, and/or person's environmental control system；
The observation system is used to observe test organization and culture medium after culture；
The analysis system obtains detection data for being analyzed according to the sensing chip；
The control system is precisely controlled drive system, realizes the control to culture medium flow velocity and flow direction in fluid channel；
The environmental control system provides suitable temperature, humidity and gas concentration lwevel for biological 3D printing class loading growth.
6. system according to claim 1, which is characterized in that the drive system includes display control unit, control dress
It sets and gas circuit distributor；The control device is connected with the display control unit, gas circuit distributor respectively；
The display control unit allows the user to multiple Working moulds according to viewing display for showing multiple operating modes
Formula is selected；
The control device is used for the operating mode selected according to user, controls the gas circuit distributor and works.
7. system according to claim 1, which is characterized in that the connection pedestal matches the organ chip and uses, institute
Use can be combined by connection type and the drive system by stating connection pedestal, or connection pedestal is combined into drive system group
One entirety carry out using.
8. system according to claim 1, which is characterized in that the system also includes: more new system, the drive system
It is connected by the more new system with the organ chip, the drive system is also used to drive described in the update system update
Culture medium in organ chip.
9. it is a kind of suitable for biological tissue culture and real-time monitoring method characterized by comprising
The biological 3D printing class loading of test/test is constructed using biological 3D printer；Wherein, the 3D printing class loading conduct
Test organization can directly be printed upon in organ chip culturing room, or be printed upon on transhipment unit and be placed into organ chip culture again
It is indoor；
By receiving the operating mode of user's selection, according to the selected operating mode culture of user in the organ chip
The test organization；
In incubation, the test organization is detected, detection data is obtained.
10. according to the method described in claim 9, it is characterized in that, it is described by receive user selection operating mode, according to
The test organization of the selected operating mode culture of user in the organ chip, comprising:
After the operating mode for receiving user's selection, by controlling positive pressure gas source, negative pressure gas source, just to the output of gas circuit distributor
Pressure, negative pressure gas flow, so that the gas passage that positive pressure, negative pressure lead to organ chip is made culture medium movement by gas circuit distributor.
11. according to the method described in claim 10, it is characterized in that, by receive user selection operating mode, according to
After the selected operating mode culture in family is the test organization in the organ chip the step of, the method is also wrapped
It includes:
It is determining that the culture medium in organ chip stores in bubble, according to bubble position, is adjusting current operation mode.
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| CN113180879A (en) * | 2021-05-24 | 2021-07-30 | 上海大学 | Biological tissue preparation system and method |
| CN116536155A (en) * | 2023-05-17 | 2023-08-04 | 扬州大学 | Chip culture device and application method thereof |
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