CN110331097A - The integrated multi-modal movement Three Dimensional Cavities intestines organ chip of enteron aisle and method - Google Patents
The integrated multi-modal movement Three Dimensional Cavities intestines organ chip of enteron aisle and method Download PDFInfo
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- CN110331097A CN110331097A CN201910713193.8A CN201910713193A CN110331097A CN 110331097 A CN110331097 A CN 110331097A CN 201910713193 A CN201910713193 A CN 201910713193A CN 110331097 A CN110331097 A CN 110331097A
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- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/10—Hollow fibers or tubes
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- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
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- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/10—Perfusion
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- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
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- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
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- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
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- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
Present disclose provides a kind of multi-modal movement Three Dimensional Cavities intestines organ chip of integrated enteron aisle and methods, including the cavity porous membrane for cultivating differentiation enterocyte, support film is provided on the outside of cavity porous membrane, the support film is connect with gas passage, pass through the import/exhaust of gas passage, support film movement is driven, and then makes the cavity porous membrane deformation occurs, to simulate tonic contraction movement；It is provided with the cavity of housing region on the outside of cavity porous membrane, culture solution perfusion region can be provided for the cavity porous membrane, being formed has the axial micromechanics environment with circumferential shear stress；Circumference is embedded with multiple closed magnetic bands on cavity porous membrane, and closed magnetic band can generate magnetic field as the solenoid being movably arranged outside cavity is powered, to make the cavity porous membrane corresponding position that radial contraction movement occur.
Description
Technical field
The disclosure belongs to microcomputer electrical engineering and field of biomedicine technology, and in particular to a kind of integrated multi-modal movement of enteron aisle
Three Dimensional Cavities intestines organ chip and method.
Background technique
Only there is provided background technical informations relevant to the disclosure for the statement of this part, it is not necessary to so constitute first skill
Art.
Currently, the new oral drug screening to disease treatment depends on animal model.New drug enter market before,
After need to carrying out curative effect and safety evaluation by animal model, it just can further implement the clinical trial of the mankind.Based on animal mould
The drug screening of type is at high cost, and time-consuming, and there are dispute of ethic；And by not being suitable in the drug of test more than 95%
The mankind, or even to mankind's toxic side effect.Enterocyte metabolic capability, absorption efficiency and cross-film between this result and species
The otherness of permeability is related.
Due to the presence of problem above, the pass of people is increasingly caused based on the in vitro enterocyte of the mankind/tissue/organ model
Note.Occur the products such as enterocyte culture systems, but the artificial intestines organ of the product of these early stages and subsequent research and development successively
And there is very big difference in body intestines organ.
The micro-fluidic device (Microfluidic device) developed by MEMS (MEMS) is that simulation is in vitro
The micro- physiology of cell/tissue/organ/Microphysical environment provides possibility, the organ chip that is further derived (OOC,
It Organ-on-a-chip is) accurate control multiple parameters, the behavioral activity of single/multiple organ of the in-vitro simulated mankind, mechanical spy
Property and physiological reaction provide possibility.Based on OOC technology, the micro-fluidic core of lung function will be simulated by Ingber and other seminars
Piece uses the differentiation culture in intestines organ instead, can be realized enterocyte-bacteria paragenesis culture and probes into pathogenic bacteria to the shadow of intestines pattern
It rings.
Understand according to inventor, these researchs are realized to explore in-vitro simulated intestines organ dysfunction and drug screening be made that it is important
Contribution, the enteron aisle of acquisition opposite Transwell model in each index of pattern and function has major progress, but still exists
Following aspect is insufficient:
It can only realize two-dimensional surface cell culture, when simulating the shear stress of microfluid, only be able to achieve axial shear stress mould
It is quasi-；Living body enteron aisle is morphologically a kind of three-dimensional cavity structure, thus lacks circumferential shear stress of the fluid in cavity.
The only single tonic contraction mode of analog cannot simulate the multi-modal forms of motion of enteron aisle (for example, offseting
The merogenesis radial contraction and promote chyme/drug peristaltic wave that change/absorption function plays a major role).
Summary of the invention
The disclosure to solve the above-mentioned problems, proposes a kind of multi-modal movement Three Dimensional Cavities intestines organ chip of integrated enteron aisle
And method, the disclosure can be realized in vitro simulation mankind's intestines organ, integrate more mechanical parameters, can differentiate closer to body intestines
Pattern, intestinal epithelial barrier and the digestive function of road organ.
According to some embodiments, the disclosure is adopted the following technical scheme that
A kind of multi-modal movement Three Dimensional Cavities intestines organ chip of integrated enteron aisle, comprising:
For cultivating the cavity porous membrane of differentiation enterocyte, it is thin that support is provided on the outside of the cavity porous membrane
Film, the support film are connect with gas passage, by the import/exhaust of the gas passage, drive support film movement, in turn
Making the cavity porous membrane, deformation occurs, to simulate tonic contraction movement；
The cavity of housing region is provided on the outside of the cavity porous membrane, it can be porous thin for the cavity
Film provides culture solution perfusion region, and being formed has the axial micromechanics environment with circumferential shear stress；
Circumference is embedded with multiple closed magnetic bands on the cavity porous membrane, and the closed magnetic band can be with removable outside cavity
The solenoid of dynamic setting, which is powered, generates magnetic field, to make the cavity porous membrane corresponding position that radial contraction movement occur.
In above-mentioned design scheme, by the cooperation between each element, cavity porous membrane can be made to generate tonicity and received
Contracting movement, the movement of merogenesis radial contraction and the axial multi-modal movement with circumferential shear stress are simulated；Joined according to more Mechanical courses
Number, explores the relationship of itself and intestines organ metabolic capability, absorption efficiency and transmembranal penetration rate.
As selectable embodiment, metabolism liquid collection channel is additionally provided on the outside of the cavity porous membrane, to receive
Collect the metabolism liquid of enterocyte.
As selectable embodiment, the cavity porous membrane is that flexible polydimethyl siloxane material is prepared into
It arrives, 3 μm~5 μm of thickness.
As selectable embodiment, the closed magnetic band is endless tape, by magnetic-particle and cavity porous membrane
It is formed by curing, is evenly spaced on cavity porous membrane surface after being sufficiently mixed.
As selectable embodiment, the solenoidal power-on voltage is adjustable, by adjusting the power of voltage, changes
Varying magnetic field intensity realizes the radial contraction of magnetic rings band difference amplitude, by control the solenoidal axial movement position with
Movement speed realizes the simulation of enteron aisle different location, different conditions.
As selectable embodiment, the gas passage includes two, and is set to the cavity porous membrane
Two sides, and the cavity porous membrane same material, by corona treatment bonding packaging, length and thickness and the cavity are porous
Film is consistent.
As selectable embodiment, cavity porous membrane medial and lateral can cultivate enterocyte, utilize intestines
Organ chip mark specifies albumen, carries out the morphology characterization of Laser Scanning Confocal Microscope, compares the formation state of intestinal villi；Pass through note
The barrier of intestinal epithelial cells monolayer for recording cross-film resistive technologies analysis inside, reflects the degree of enterocyte gap connection indirectly.
The preparation method of above-mentioned Three Dimensional Cavities intestines organ chip, comprising the following steps:
Prepare cavity porous membrane: one layer of SU-8 negative photoresist of spin coating on silicon wafer is cooled to room temperature after vacuum drying,
Photomask is contacted and exposed with coating silicon wafers, heating exposure chip, and be heat-treated or postexposure bake, micro-pillar array
Then by by silicon wafer time of infusion in developer solution, then rinsed with water and dry in air and develop, it will be flexible
Polydimethyl siloxane material is spin-coated in micro-pillar array, is removed after cured, and film is enclosed Teflon hose and attaches a circle, is led to
It crosses semi-solid preparation flexibility polydimethyl siloxane material and realizes close joint；
It prepares micro- gas channel: solidifying after flexible polydimethyl siloxane material is poured into mold, the mold includes structure
Identical two parts, each part are oppositely arranged at least two protrusions, two parts, by plasma treatment, are realized
Agree with encapsulation；
Closed magnetic band is prepared, by Fe3O4Particle is sufficiently mixed with flexible polydimethyl siloxane material, then by it with rotation
It is applied to the uniform acrylic board in surface, is waited after cured, removing activates cavity flexibility polydimethyl siloxane material with Plasma
Film surface, by the Fe after removing3O4Thin magnetic film uniformly sticks its surface at equal intervals；
The three parts being prepared are completed into assembling by key and technique.
The working method of above-mentioned Three Dimensional Cavities intestines organ chip, comprising the following steps:
By the import/exhaust of the gas passage, support film movement is driven, and then the cavity porous membrane occurs
Deformation, to simulate and analyze tonic contraction；
Or/and the flow rate by changing the nutrient solution, being formed has the axial micromechanics ring with circumferential shear stress
The relationship of flow rate of liquid and shear stress is simulated and is analyzed in border；
Or/and it by changing the voltage and solenoidal movement speed that solenoid is powered, simulates and analyzes enteron aisle
Radial contraction and peristaltic wave.
As selectable embodiment, the simulation and when analyzing tonic contraction radial contraction, preparing with it is living
Under conditions of the cavity film of body intestinal tissue same stiffness and thickness, magnetic field strength and PDMS film deformation are established using simulation software
Calibration relationship, change the deformation that solenoidal voltage finds cavity film corresponding with deformation, it is thin to cavity by solenoid
Film realizes merogenesis radial contraction according to the frequency of setting；
The simulation and when analyzing peristaltic wave, alternately attract to closed magnetic band and release, the speed magnetic of peristaltic wave
Property annulus gap and the time difference of wave crest two-by-two calculate, determine the pass between solenoidal friction speed and cavity wriggling velocity of wave
System.
As selectable embodiment, the simulation and when analyzing tonic contraction utilizes two wing passages to realize air pressure
Oriented film deformation determines the relationship of different air pressures and deformation.
As selectable embodiment, the simulation and when analyzing the relationship of flow rate of liquid and shear stress, according to living body
Liquid flow rate in enteron aisle calculates ideally inner shaft shear stress, which is imported simulation software, backward simulation
The flowing velocity of culture solution needed for cavity intestines organ chip out.
Compared with prior art, the disclosure has the beneficial effect that
The drawbacks of disclosure lacks the multi-modal motor pattern of internal enteron aisle for it, proposes that one kind is done more physical exercises parameter simulation
Three Dimensional Cavities intestines organ chip, realize the pattern, intestinal epithelial barrier and digestive function closer to living body intestines organ in vitro；
The tonic contraction, merogenesis radial contraction, peristaltic wave, the circumferential multi-modal simulation with axial shear stress of simulation enteron aisle can be achieved；
The influence that more mechanical parameters form enteron aisle pattern, function is really disclosed under bionical state, is the research etc. of intestines organ chip
Provide help.
Detailed description of the invention
The Figure of description for constituting a part of this disclosure is used to provide further understanding of the disclosure, and the disclosure is shown
Meaning property embodiment and its explanation do not constitute the improper restriction to the disclosure for explaining the disclosure.
Fig. 1 (a) be the disclosure cavity porous membrane under the air pressure of 10kPa caused stress distribution schematic diagram；
Fig. 1 (b) is maximum strain and atmospheric pressure value schematic diagram in a linear relationship in the air pressure range of 5kPa to 200kPa；
Fig. 2 is the preparation flow figure of the cavity porous membrane of the disclosure；
Fig. 3 is the long organ chip preparation flow schematic diagram of Three Dimensional Cavities of the disclosure；
Fig. 4 is the structural schematic diagram of the disclosure.
Specific embodiment:
The disclosure is described further with embodiment with reference to the accompanying drawing.
It is noted that following detailed description is all illustrative, it is intended to provide further instruction to the disclosure.Unless another
It indicates, all technical and scientific terms used herein has usual with disclosure person of an ordinary skill in the technical field
The identical meanings of understanding.
It should be noted that term used herein above is merely to describe specific embodiment, and be not intended to restricted root
According to the illustrative embodiments of the disclosure.As used herein, unless the context clearly indicates otherwise, otherwise singular
Also it is intended to include plural form, additionally, it should be understood that, when in the present specification using term "comprising" and/or " packet
Include " when, indicate existing characteristics, step, operation, device, component and/or their combination.
In the disclosure, term for example "upper", "lower", "left", "right", "front", "rear", "vertical", "horizontal", " side ",
The orientation or positional relationship of the instructions such as "bottom" is to be based on the orientation or positional relationship shown in the drawings, only to facilitate describing this public affairs
The relative for opening each component or component structure relationship and determination, not refers in particular to either component or element in the disclosure, cannot understand
For the limitation to the disclosure.
In the disclosure, term such as " affixed ", " connected ", " connection " be shall be understood in a broad sense, and indicate may be a fixed connection,
It is also possible to be integrally connected or is detachably connected；It can be directly connected, it can also be indirectly connected through an intermediary.For
The related scientific research of this field or technical staff can determine the concrete meaning of above-mentioned term in the disclosure as the case may be,
It should not be understood as the limitation to the disclosure.
As shown in figure 4, a kind of Three Dimensional Cavities intestines organ chip of the multi-modal movement simulation of integrated enteron aisle, the chip can be used for
Pattern differentiation, function are formed and research, including seven parts coexist in bacterium colony, is respectively as follows: cavity porous membrane (1), is spaced apart
Magnetic rings band (2), removable solenoid (3), metabolism liquor collecting device (4), culture solution perfusion device (5), two sides gas it is logical
Road (6) (7) and support film (8).Wherein, cavity porous membrane (1) is the core of entire chip, and cultivating inside and outside it can break up
For enterocyte, two sides are provided with gas passage (6) (7), are cultured liquid perfusion device (5) encirclement.Endless tape (2) insertion
On cavity porous membrane (1) surface, and equally it is distributed；Removable solenoid (3) can be axial along cavity porous membrane (1)
It is mobile, and after its energization, radial contraction can be brought it about by generating magnetic field.
Specifically, in the present embodiment, the basis material of cavity porous membrane selects modulated rigidity, good biocompatibility
Flexible dimethyl silicone polymer (PDMS) material, 2 μm of chip feature sizes (i.e. the diameter of porous membrane), cavity porous membrane
3 μm~5 μm of thickness, the length 38mm, width 13mm, thickness 1mm~2mm of entire Three Dimensional Cavities intestines organ chip.
Certainly, in other embodiments, above-mentioned size all can be changed and be adjusted, this belongs to those skilled in the art
The simple replacement being readily apparent that ought to belong to the protection scope of the disclosure.
Endless tape (2) by partial size 50nm~150nm magnetic-particle (such as Fe3O4) with PDMS be sufficiently mixed after solidify shape
At being evenly spaced in cavity porous membrane (1) surface；After removable solenoid (3) are powered, by adjusting the strong of voltage
It is weak, change magnetic field strength, realizes the radial contraction of magnetic rings band difference amplitude.In the present embodiment, 500 μ of magnetic rings bandwidth
M, line space design 1.5mm.Likewise, in other embodiments, above-mentioned size all can be changed and be adjusted, this belongs to this
The simple replacement that field technical staff is readily apparent that ought to belong to the protection scope of the disclosure.
Wave axially wriggling is presented to shrink it, removable solenoid (3) should be kept according to enteron aisle bionic principle
The axial movement of 0.5cm/min~2cm/min.
Two sides gas passage (6) (7) produces cavity porous membrane integrally by micro- air pump import/exhaust, through support film (8)
Raw deformation, the simulation for enteron aisle tonic contraction.Two sides gas passage (6) (7) side length and thickness and chip keep one
It causes, width can choose 5mm；It, which is prepared, passes through PDMS aluminum alloy mould mold twice, complete with corona treatment bonding packaging
At.
Liquor collecting device (4) and culture solution perfusion device (5) are metabolized for the liquid perfusion of entire cavity chip and sealing.
Aluminum dipping form of two kinds of devices through milling process completes preparation, by with corona treatment bonding technology, by support film (8) with
Cavity porous membrane (1) realizes sealing.The cavity body structure of peripheral sealing, it can be achieved that culture solution slow perfusion, in Three Dimensional Cavities
The inside and outside formation of porous membrane axially with circumferential shear stress, forms similar micromechanics environment with living body enteron aisle.
Metabolism liquor collecting device is set to film outside, and the differentiation with culture solution perfusion device is that perfusion device is whole
Top half on the outside of a cavity chip is metabolized collection device in the lower half portion of entire cavity chip.It is metabolized liquor collecting device
(4) film separates between culture solution perfusion device (5).
By above structure, more mechanical parameter stimulations are formed, is cultivated in cavity porous membrane (8) and is divided into similar people's intestines
The pattern and enterocyte type in road, distribution.In conjunction with existing intestines organ chip (Transwell model and two-dimentional intestines organ
Chip) specific protein (such as actin, Fibronectin) is marked respectively, the morphology characterization of Laser Scanning Confocal Microscope is carried out, it is smaller
The formation state of intestinal villus.
The enterocyte of portion's culture in the cavity can analyze barrier of intestinal epithelial cells monolayer by record cross-film resistive technologies
, reflect the degree of enterocyte gap connection indirectly.
The enterocyte can will mark the culture of monosaccharide by culture solution perfusion device (5) containing fluorescein FD20
Liquid flows slowly into cavity perforated membrane, and carries out fluorescein statistics by metabolism liquor collecting device (4), for analyzing enterocyte
Digestion, absorption and infiltration function.
Probiotics can be passed through in the cavity porous membrane (1), for constructing enteron aisle/bacterium coexisted environment, and be compared
The influence of the pattern, function to chip coexists compared with both analyses.Bar class pathogenic bacteria are passed through in the cavity porous membrane (1), use
In the enteritis class disease model (such as Crow grace) that building is common；Observation analysis is there are the class living body intestinal environments of more mechanical parameters
In, pathogenic bacteria infect situation.
The preparation process of Three Dimensional Cavities intestines organ chip mainly includes Three Dimensional Cavities film, micro- gas channel, Fe3O4Magnetic rings
Three parts of band.
It specifically includes:
Cavity film preparation: firstly, with the SU-8 negative photoresist of one layer of 30 μ m-thick of 2500r/min spin coating on silicon wafer.
It is dried in vacuo 20 minutes at 95 DEG C and is then cooled to room temperature.Then, photomask is contacted and is exposed with coating silicon wafers.Then,
It was heat-treated by heating exposure chip 10 minutes at 95 DEG C or postexposure bake.Micro-pillar array is then by by silicon
Piece impregnates 10 minutes in developer solution, is then developed with water rinsing and in air drying.By PDMS (mass ratio 20:1) with
3000r/min is spin-coated in micro-pillar array, after cured by its careful removing.Film is enclosed into Teflon hose and attaches a circle, is led to
It crosses semi-solid preparation PDMS and realizes close joint.
Specifically as shown in Fig. 2, being divided into:
(A) one layer of 20 μm of SU-8 photoresist are got rid of in silicon chip surface；
(B) uv-exposure is handled；
(C) development operation；
(D) SU-8 microtrabeculae template is prepared；
(E) template being formed by curing；
(F) sol evenning machine gets rid of one layer of PDMS；
(G) PDMS film is removed；
(H) it cuts out as rectangular film；
(I) plasma treatment, be bonded to cavity porous membrane.
(2) micro- gas channel preparation: micro- gas channel includes about two identical parts, and every part is all by PDMS (quality
Than 10:1) pour into aluminium grinding tool after solidify.Aluminum die is prepared with milling process, and there are two rectangular-shape protrusion (height 150 for band
μm, width is 1000 μm).Two parts agree with encapsulation by plasma treatment, realization.(see Fig. 3).
It specifically includes:
(A) aluminium-making mould of milling process preparation prepares chip two symmetric parts up and down；
(B) cavity perforated membrane is embedded in two symmetric parts up and down；
(C) after corona treatment, bonding packaging；
(D) the perforated membrane two sides for peeling off support membrane insertion；
(E) corona treatment；
(F) bonding encapsulation.
(3)Fe3O4Magnetic rings band: by the Fe of diameter 100nm3O4Particle is filled with PDMS (mass ratio 20:1) by stirring instrument
Divide mixing (the power 60W of stirring instrument runs more than 4h), it is then spin-coated to the uniform acrylic in surface with 4000r/min
Plate waits after cured, careful to remove.Cavity PDMS film surface is activated with Plasma, by the Fe after removing3O4Thin magnetic film, etc.
Uniformly stick its surface in interval.Finally, three parts are completed assembling by key and technique.
In above-mentioned preparation process, the kinematic parameter (such as power, time) of the equipment, equipment that are applied to, material and each
The parameter (such as proportion, partial size) of material, technological parameter (such as temperature) etc., can be replaced in other embodiments
It changes, these belong to what those skilled in the art were readily apparent that, utilize the protection scope for belonging to the disclosure.
The more mechanical parameters of cavity intestines organ integrated chip of formation can be used for the realization of multi-modal movement simulation, research
Content may include:
(1) to simulate enteron aisle merogenesis radial contraction, it is strong simulation enteron aisle merogenesis radial contraction and wriggling: to first have to searching magnetic field
Relationship between degree and strain.Under conditions of it is assumed that the cavity film with living body intestinal tissue same stiffness and thickness can be prepared,
The calibration relationship of magnetic field strength Yu PDMS film deformation (draw ratio of diameter) is established using Comsol numerical simulation.Then, change
Become the deformation that solenoidal voltage finds cavity film corresponding with deformation.Finally, by solenoid to cavity film according to
The frequency of 0.13Hz~0.18Hz realizes merogenesis radial contraction.To simulate the peristaltic wave that enteron aisle promotes chyme/drug forward, we
Equally distributed Fe3O4 magnetic rings tape alternation will be attracted and be discharged.The speed of peristaltic wave magnetic rings band gap and wave two-by-two
The time difference at peak calculates.Set solenoidal friction speed (0.5cm/s, 1.0cm/s, 1.5cm/s, 2cm/s) calibration and cavity
Relationship between wriggling velocity of wave.
(2) tonic contraction simulated implementation: tonic contraction simulation will be using gas similar with two-dimentional intestines organ chip
Stretching technique is realized and (realizes the deformation of air pressure hoist film using two wing passages).The difference is that we towards be cavity draw
Deformation is stretched, so it is different that Ansys emulates obtained application atmospheric pressure value under identical deformation.
(3) it simulation microfluid flow velocity and shear stress relationship: firstly, by the liquid flow rate in living body enteron aisle, calculates
Ideally inner shaft shear stress；Secondly, the numerical value is imported Comsol software, backward simulation goes out cavity intestines organ chip institute
The flowing velocity of the culture solution needed.
Parameter used in the studies above process can be replaced as the case may be.These belong to those skilled in the art
What member was readily apparent that, utilize the protection scope for belonging to the disclosure.
The foregoing is merely preferred embodiment of the present disclosure, are not limited to the disclosure, for the skill of this field
For art personnel, the disclosure can have various modifications and variations.It is all within the spirit and principle of the disclosure, it is made any to repair
Change, equivalent replacement, improvement etc., should be included within the protection scope of the disclosure.
Although above-mentioned be described in conjunction with specific embodiment of the attached drawing to the disclosure, model not is protected to the disclosure
The limitation enclosed, those skilled in the art should understand that, on the basis of the technical solution of the disclosure, those skilled in the art are not
Need to make the creative labor the various modifications or changes that can be made still within the protection scope of the disclosure.
Claims (10)
1. a kind of multi-modal movement Three Dimensional Cavities intestines organ chip of integrated enteron aisle, it is characterized in that: including:
For cultivating the cavity porous membrane of differentiation enterocyte, the cavity porous membrane outside is provided with support film,
The support film is connect with gas passage, by the import/exhaust of the gas passage, drives support film movement, and then make
Deformation occurs for the cavity porous membrane, to simulate tonic contraction movement；
It is provided with the cavity of housing region on the outside of the cavity porous membrane, can be mentioned for the cavity porous membrane
For culture solution perfusion region, being formed has the axial micromechanics environment with circumferential shear stress；
Circumference is embedded with multiple closed magnetic bands on the cavity porous membrane, and the closed magnetic band can be set with removable outside cavity
The solenoid set, which is powered, generates magnetic field, to make the cavity porous membrane corresponding position that radial contraction movement occur.
2. a kind of integrated multi-modal movement Three Dimensional Cavities intestines organ chip of enteron aisle as described in claim 1, it is characterized in that: described
Metabolism liquid collection channel is additionally provided on the outside of cavity porous membrane, to collect the metabolism liquid of enterocyte.
3. a kind of integrated multi-modal movement Three Dimensional Cavities intestines organ chip of enteron aisle as described in claim 1, it is characterized in that: described
Cavity porous membrane is that flexible polydimethyl siloxane material is prepared, 3 μm~5 μm of thickness.
4. a kind of integrated multi-modal movement Three Dimensional Cavities intestines organ chip of enteron aisle as described in claim 1, it is characterized in that: described
Closed magnetic band be endless tape, be formed by curing after being sufficiently mixed by magnetic-particle with cavity porous membrane, be evenly spaced in
On cavity porous membrane surface.
5. a kind of integrated multi-modal movement Three Dimensional Cavities intestines organ chip of enteron aisle as described in claim 1, it is characterized in that: described
Solenoidal power-on voltage is adjustable, by adjusting the power of voltage, changes magnetic field strength, realizes magnetic rings band difference amplitude
Radial contraction realizes enteron aisle different location, different conditions by controlling the solenoidal axial movement position and movement speed
Simulation.
6. a kind of integrated multi-modal movement Three Dimensional Cavities intestines organ chip of enteron aisle as described in claim 1, it is characterized in that: described
Gas passage includes two, and is set to the two sides of the cavity porous membrane, and the cavity porous membrane same material, by etc.
Gas ions handle bonding packaging, and length and thickness are consistent with the cavity porous membrane.
7. a kind of integrated multi-modal movement Three Dimensional Cavities intestines organ chip of enteron aisle as described in claim 1, it is characterized in that: described
Cavity porous membrane medial and lateral can cultivate enterocyte, specify albumen using intestines organ chip mark, and it is burnt aobvious to carry out copolymerization
The morphology characterization of micro mirror compares the formation state of intestinal villi；Pass through the enteric epithelium screen of record cross-film resistive technologies analysis inside
Hinder function, reflects the degree of enterocyte gap connection indirectly.
8. the preparation method of Three Dimensional Cavities intestines organ chip of any of claims 1-7, feature and be: including with
Lower step:
Prepare cavity porous membrane: one layer of SU-8 negative photoresist of spin coating on silicon wafer is cooled to room temperature, by light after vacuum drying
Mask is contacted and is exposed with coating silicon wafers, heating exposure chip, and is heat-treated or postexposure bake, micro-pillar array are subsequent
By by silicon wafer time of infusion in developer solution, then rinsed with water and dry in air and develop, by flexible poly- two
Methylsiloxane material is spin-coated in micro-pillar array, is removed after cured, and film is enclosed Teflon hose and attaches a circle, passes through half
Solidify flexible polydimethyl siloxane material and realizes close joint；
It prepares micro- gas channel: solidifying after flexible polydimethyl siloxane material is poured into mold, the mold includes that structure is identical
Two parts, each part is at least two protrusions, and two parts are oppositely arranged, and by plasma treatment, realization agrees with
Encapsulation；
Closed magnetic band is prepared, by Fe3O4Particle is sufficiently mixed with flexible polydimethyl siloxane material, then by it to be spin-coated to
The uniform acrylic board in surface waits after cured, removing, activates cavity flexibility polydimethyl siloxane material film with Plasma
Surface, by the Fe after removing3O4Thin magnetic film uniformly sticks its surface at equal intervals；
The three parts being prepared are completed into assembling by key and technique.
9. the working method of Three Dimensional Cavities intestines organ chip of any of claims 1-7, it is characterized in that: including following
Step:
By the import/exhaust of the gas passage, support film movement is driven, and then makes the cavity porous membrane that shape occur
Become, to simulate and analyze tonic contraction；
Or/and the flow rate by changing the nutrient solution, being formed has the axial micromechanics environment with circumferential shear stress,
Simulate and analyze the relationship of flow rate of liquid and shear stress；
Or/and it is radial by changing the voltage and solenoidal movement speed that solenoid is powered, simulating and analyzing enteron aisle generation
Contraction and peristaltic wave.
10. working method as claimed in claim 9, it is characterized in that: the simulation and when analyzing tonic contraction radial contraction,
Under conditions of preparing the cavity film with living body intestinal tissue same stiffness and thickness, using simulation software establish magnetic field strength with
The calibration relationship of PDMS film deformation changes the deformation that solenoidal voltage finds cavity film corresponding with deformation, passes through spiral shell
Spool realizes merogenesis radial contraction according to the frequency of setting to cavity film；
Or,
The simulation and when analyzing peristaltic wave, alternately attract to closed magnetic band and release, the speed magnetic rings of peristaltic wave
The time difference of band gap and wave crest two-by-two calculates, and determines the relationship between solenoidal friction speed and cavity wriggling velocity of wave；
Or,
The simulation and when analyzing tonic contraction, realizes the deformation of air pressure hoist film using two wing passages, determines different air pressures
With the relationship of deformation；
Or,
When the relationship for simulating and analyzing flow rate of liquid and shear stress, according to the liquid flow rate in living body enteron aisle, calculate
The numerical value is imported simulation software, training needed for backward simulation goes out cavity intestines organ chip by ideally inner shaft shear stress
The flowing velocity of nutrient solution.
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| CN114107175A (en) * | 2021-11-22 | 2022-03-01 | 北京大学 | Organoid culture method for inducing growth of intestinal organoids by mechanical stretching |
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| CN114107175B (en) * | 2021-11-22 | 2024-01-02 | 北京大学 | Organoid culture method for inducing growth of intestinal organoids by mechanical stretching |
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