CN106811413A - Multiple organ chip based on microflow control technique and preparation method thereof - Google Patents
Multiple organ chip based on microflow control technique and preparation method thereof Download PDFInfo
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- CN106811413A CN106811413A CN201510850055.6A CN201510850055A CN106811413A CN 106811413 A CN106811413 A CN 106811413A CN 201510850055 A CN201510850055 A CN 201510850055A CN 106811413 A CN106811413 A CN 106811413A
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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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/16—Microfluidic devices; Capillary tubes
-
- 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
- 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
- C12M35/08—Chemical, biochemical or biological means, e.g. plasma jet, co-culture
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
- G01N33/5058—Neurological cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
- G01N33/5064—Endothelial cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
- G01N33/5067—Liver cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2503/00—Use of cells in diagnostics
- C12N2503/02—Drug screening
Abstract
The invention provides a kind of multiple organ chip based on microflow control technique and preparation method thereof, the micro-fluidic chip is mainly made up of upper strata chip, carbonic acid adipose membrane and lower layer chip.Its preparation method is：Upper strata chip and carbonic acid adipose membrane oxygen plasma carry out sealing-in；For bonding after levels chip upper surface dips PDMS, bonding of aliging is carried out with the irreversible upper strata chip for being sealed with carbonic acid adipose membrane.Advantages of the present invention is：Using the method that multi-layer micro-fluidic chips are integrated, the culture unit of various kinds of cell is combined, according to the sequencing run between intracorporeal organ, by the model assembling of various organs and structure on same chip, simple and efficient realizes the united chip construction method of multiple organ, and can effectively be split between organ, it is easy to control respectively and observation between each organ.
Description
Technical field
Simulation the present invention relates to microfluidic chip technology to be applied to external many tissues and organ is led with application
A kind of domain, and in particular to multiple organ chip based on microflow control technique and preparation method thereof.
Background technology
During medical research and new drug development, suitable model is the importance studied.Just
Animal experimental model is generated, meanwhile, in order to study influence of the new drug to human body, scientist starts in culture
Human body cell is cultivated in ware, look at these new drugs to human body cell whether toxic side effect.But animal and
Mankind's isolated cells can not hold the gate of reagent for the mankind.According to statistics, can pass through for three phases clinical
The medicine of experiment only has 10.8%, main cause because curative effect is bad, even to human body toxic side effect,
This is huge waste.In order to solve this problem, US National health research center (NIH) in 2012,
Food and drug administration (FDA) and ARPA (DARPA) combine
Initiate the R＆D work of " organs-on-chips " (human organ chip).
Microfluidic chip technology has been obtained extensively as a technology for developing rapidly in biomedical sector
General application, and match because of itself and cell size, nearly physiological microenvironment and space-time controllability, it is easy to
The application of cellular level is realized by flexible structure design and various preparation method and is increasingly becoming new
The important technical and platform of cytology research.At present, single organization is carried out using microfluidic chip technology
Or the simulation work of organ has launched, but Various Tissues or organ are assembled on same chip
With the related work simulated also in blank stage.Therefore, multiple organ is carried out using microfluidic chip technology
Simulation has great application prospect with bionical in biological study and medicine research and development.
The content of the invention
The invention provides a kind of multiple organ chip based on microflow control technique and preparation method thereof, particular for
With chip simulation associated with the two or more organ with function continuity in vivo.
A kind of multiple organ chip based on microflow control technique, the chip mainly includes three-decker：Upper strata chip,
Middle level carbonic acid adipose membrane and lower layer chip；There is micropore, the cell cultivated in the chip of upper strata on the carbonic acid adipose membrane of middle level
Produced metabolite can pass through micropore permeation in lower layer chip, and then stimulate what is cultivated in lower layer chip
Cell or tissue model, so that the function continuity in analogue body between multiple organ；
The upper strata chip includes the outlet of upper cell entrance, upper cell passage and upper cell；Upper strata is thin
Born of the same parents' entrance and upper cell outlet are connected by upper cell passage；
The lower layer chip includes that underlying collagenous entrance, underlying collagenous passage, lower floor's cell entry, lower floor are thin
Born of the same parents are exported and lower floor's cell passage, and underlying collagenous entrance and underlying collagenous passage are connected, underlying collagenous passage and
Lower floor's cell passage is directly connected, and lower floor's cell entry and lower floor's cell outlet are connected by lower floor's cell passage；
After chip entirety sealing-in, upper cell passage and lower floor's cell passage mutually correspond to weight in position
Close.
The micro-fluidic chip that the present invention is provided, the design of whole chip can side by side sealing-in in sufficiently large cleaned glass
To increase flux, number can be between 1-100 for bottom surface.
A kind of preparation method of the multiple organ chip based on microflow control technique, the lower surface and carbonic acid of upper strata chip
Adipose membrane by UV activation 1 hour, silanization treatment 30 minutes, oxygen plasma carries out sealing-in；For bonding
The PDMS of levels chip is monomer and ratio of initiator is 20:1 PDMS polymer, in lower layer chip
After surface dips PDMS, bonding of aliging is carried out with the irreversible upper strata chip for being sealed with carbonic acid adipose membrane.
The upper strata chip and lower layer chip are the PDMS polymer of light transmission ventilation, PDMS monomers with draw
Hair agent ratio is 6:1.
A kind of application of the multiple organ chip based on microflow control technique, upper strata chip is used for culture hepatocyte, institute
It is the internal pro-drug for needing and being activated by hepatic metabolism with medicine；Lower layer chip is used to build external blood
Whether whether brain BARRIER MODEL, can be by blood-brain barrier and right by the medicine of hepatic metabolism for evaluating
The function of blood-brain barrier has destructive or protective effect；The collagen porch of lower layer chip can be used for simultaneously
Collection to passing through the liquid of blood-brain barrier in lower layer chip.
Advantages of the present invention is：Using the method that multi-layer micro-fluidic chips are integrated, by the culture list of various kinds of cell
Unit combines, and according to the sequencing run between intracorporeal organ, the model assembling of various organs and structure is existed
On same chip, simple and efficient realizes the united chip construction method of multiple organ, and can between organ
Effectively split, be easy to control respectively and observation between each organ.
Brief description of the drawings
Fig. 1 micro-fluidic chip overall structure diagrams provided by the present invention；
The flow chart of the preparation method of Fig. 2 micro-fluidic chips provided by the present invention；
Wherein, 1 upper strata chip, 2 upper cell entrances, 3 upper channels, the outlet of 4 upper cells, 5 middle levels
Carbonic acid adipose membrane；6 lower layer chips, 7 underlying collagenous entrances, 8 underlying collagenous passages, 9 lower floor's cell entries,
10 lower floor's cell outlets, 11 lower floor's cell passages；12 cleaned glass.
Specific embodiment
The following examples will be further described to the present invention, but not thereby limiting the invention.
Embodiment 1
A kind of multiple organ chip based on microflow control technique, as shown in Figure 1：It is a kind of based on microflow control technique
Multiple organ microarray biochip mainly includes three-decker, upper strata chip 1, middle level carbonic acid adipose membrane 5 and lower layer chip 6；
There is micropore, the metabolite produced by cell cultivated in upper strata chip 1 can be saturating on middle level carbonic acid adipose membrane 5
Micropore permeation is crossed in lower layer chip 6, and then stimulates the cell or tissue model cultivated in lower layer chip 6.
The upper strata chip 1 includes upper cell entrance 2, upper cell passage 3 and upper cell outlet 4；
Upper cell entrance 2 and upper cell outlet 4 are connected by upper cell passage 3；
The lower layer chip 6 include underlying collagenous entrance 7, underlying collagenous passage 8, lower floor's cell entry 9,
Lower floor's cell outlet 10 and lower floor's cell passage 11, underlying collagenous entrance 7 and underlying collagenous passage 8 are connected,
Underlying collagenous passage 8 is directly connected with lower floor cell passage 11, lower floor's cell entry 9 and lower floor's cell outlet
10 are connected by lower floor's cell passage 11；
After chip entirety sealing-in, upper cell passage 3 and lower floor's cell passage 11 are mutually right in position
Should overlap.
The making of upper strata chip and lower layer chip
It is material using SU-8 3035, upper strata chip 1 and lower floor is made respectively according to traditional soft lithography process
The template of chip 6, its at the middle and upper levels chip be single tier templates, for culture hepatocyte；Lower layer chip is bilayer
Template, for building in vitro blood-brain barrier model.After the completion of template construct, poured using PDMS polymer
Template (monomer：Initiator=6:1) the PDMS blocks of upper strata chip and lower layer chip, are made respectively and are beaten with self-control
Hole device punching is standby,.
The sealing-in of overall chip
Carbonic acid adipose membrane 5 is placed on slide, UV activation 1 hour in super-clean bench is positioned over, then use silanization treatment
30 minutes, subsequent upper strata chip 1 together carried out plasma sealing-in, is subsequently placed in 80 DEG C of baking ovens and processes 30
Minute.The direct plasma sealing-in of lower layer chip 6 is on one piece of cleaned glass 12 of suitable size.Prepare again
Monomer is 20 with ratio of initiator:1 PDMS polymer, after a thin layer is smeared in the upper surface of lower layer chip,
Layers of chips is directly alignd bonding, finally overall chip is placed in 80 DEG C of baking ovens and is solidified 30 minutes, such as schemed
Shown in 2.
Claims (7)
1. a kind of multiple organ chip based on microflow control technique, it is characterised in that：The chip mainly includes three-decker：Upper strata chip (1), middle level carbonic acid adipose membrane (5) and lower layer chip (6)；There is micropore on middle level carbonic acid adipose membrane (5), metabolite in upper strata chip (1) produced by the cell of culture can pass through micropore permeation in lower layer chip (6), and then stimulate the cell or tissue model cultivated in lower layer chip (6)；
The upper strata chip (1) includes upper cell entrance (2), upper cell passage (3) and upper cell outlet (4)；Upper cell entrance (2) and upper cell outlet (4) are connected by upper cell passage (3)；
The lower layer chip (6) includes underlying collagenous entrance (7), underlying collagenous passage (8), lower floor's cell entry (9), lower floor's cell outlet (10) and lower floor's cell passage (11), underlying collagenous entrance (7) and underlying collagenous passage (8) are connected, underlying collagenous passage (8) is directly connected with lower floor's cell passage (11), and lower floor's cell entry (9) and lower floor's cell outlet (10) are connected by lower floor's cell passage (11)；
After chip entirety sealing-in, mutually correspondence overlaps in position for upper cell passage (3) and lower floor's cell passage (11).
2. according to a kind of multiple organ chip based on microflow control technique described in claim 1, it is characterised in that：Whole chip design can side by side sealing-in in sufficiently large cleaned glass bottom surface to increase flux, number can be between 1-100.
3. according to a kind of preparation method of the multiple organ chip based on microflow control technique described in claim 1, it is characterised in that follow the steps below：The lower surface and carbonic acid adipose membrane of upper strata chip by UV activation 1 hour, silanization treatment 30 minutes, oxygen plasma carries out sealing-in；For bonding the PDMS of levels chip for monomer and ratio of initiator are 20:1 PDMS polymer, after chip upper surface dips PDMS, bonding of aliging is carried out with the irreversible upper strata chip for being sealed with carbonic acid adipose membrane.
4. according to a kind of preparation method of the multiple organ chip based on microflow control technique described in claim 3, it is characterised in that：Upper strata chip and lower layer chip are the PDMS polymer of light transmission ventilation, and PDMS monomers are 6 with ratio of initiator:1.
5. according to a kind of application of the multiple organ chip based on microflow control technique described in claim 1, it is characterised in that：Upper strata chip is used for culture hepatocyte, and medicine used is the pro-drug for needing to be activated by hepatic metabolism in vivo.
6. according to a kind of application of the multiple organ chip based on microflow control technique described in claim 1, it is characterised in that：Lower layer chip is used to build in vitro blood-brain barrier model, and whether whether the medicine for evaluating by hepatic metabolism by blood-brain barrier, and can have destructive or protective effect to the function of blood-brain barrier.
7. according to a kind of application of the multiple organ chip based on microflow control technique described in claim 1, it is characterised in that：The collagen porch of lower layer chip can be used for the collection of the liquid to passing through blood-brain barrier in lower layer chip simultaneously.
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Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107828655A (en) * | 2017-11-15 | 2018-03-23 | 大连理工大学 | A kind of micro-fluidic chip and its application |
| WO2019019526A1 (en) * | 2017-07-28 | 2019-01-31 | 中科芯瑞(苏州)生物科技有限公司 | Microfluidic chip and application thereof |
| CN109517739A (en) * | 2018-12-27 | 2019-03-26 | 重庆大学 | Double organs are coupled three-layer laminated integrated chip |
| CN110106081A (en) * | 2019-05-13 | 2019-08-09 | 大连医科大学附属第一医院 | For constructing the micro-fluidic chip and construction method of brain function model of element |
| CN110117524A (en) * | 2019-04-17 | 2019-08-13 | 华南理工大学 | A kind of micro flow control chip device of hydrodynamic shear induction tumour cell cross-film migration |
| CN110408538A (en) * | 2019-07-18 | 2019-11-05 | 合肥学院 | A kind of liver chip of more lobuli hepatis integrated morphologies |
| WO2019222871A1 (en) * | 2018-05-21 | 2019-11-28 | 深圳华大生命科学研究院 | Bionic intestinal-hepatic organ chip, preparation method therefor and application thereof |
| CN110511866A (en) * | 2018-05-21 | 2019-11-29 | 深圳华大生命科学研究院 | A kind of multiple organ chip and its preparation method and application |
| CN110643502A (en) * | 2019-10-27 | 2020-01-03 | 苏州济研生物医药科技有限公司 | Single-cell microfluidic detection chip and preparation method and application method thereof |
| CN110669670A (en) * | 2019-11-14 | 2020-01-10 | 大连理工大学 | Human body micro-ecosystem chip and use method thereof |
| CN110862926A (en) * | 2018-08-27 | 2020-03-06 | 中国科学院大连化学物理研究所 | Multilayer paper chip based on microfluidic technology and construction method thereof |
| CN111269830A (en) * | 2018-12-05 | 2020-06-12 | 中国科学院大连化学物理研究所 | Multi-organ chip based on microfluidic technology and application thereof |
| CN111269831A (en) * | 2018-12-05 | 2020-06-12 | 中国科学院大连化学物理研究所 | Transparent multilayer film-sandwiched micro-fluidic chip and preparation method and application thereof |
| CN111269833A (en) * | 2018-12-05 | 2020-06-12 | 中国科学院大连化学物理研究所 | Human pancreatic island organoid model construction method based on organ chip |
| CN112553076A (en) * | 2020-12-15 | 2021-03-26 | 东南大学 | Brain organoid in vitro culture chip |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019019526A1 (en) * | 2017-07-28 | 2019-01-31 | 中科芯瑞(苏州)生物科技有限公司 | Microfluidic chip and application thereof |
| CN107828655A (en) * | 2017-11-15 | 2018-03-23 | 大连理工大学 | A kind of micro-fluidic chip and its application |
| CN110511866A (en) * | 2018-05-21 | 2019-11-29 | 深圳华大生命科学研究院 | A kind of multiple organ chip and its preparation method and application |
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| CN111971384B (en) * | 2018-05-21 | 2022-06-03 | 深圳华大生命科学研究院 | Bionic intestine-liver organ chip and preparation method and application thereof |
| WO2019222871A1 (en) * | 2018-05-21 | 2019-11-28 | 深圳华大生命科学研究院 | Bionic intestinal-hepatic organ chip, preparation method therefor and application thereof |
| CN110862926A (en) * | 2018-08-27 | 2020-03-06 | 中国科学院大连化学物理研究所 | Multilayer paper chip based on microfluidic technology and construction method thereof |
| CN111269830B (en) * | 2018-12-05 | 2022-07-15 | 中国科学院大连化学物理研究所 | Multi-organ chip based on microfluidic technology and application thereof |
| CN111269830A (en) * | 2018-12-05 | 2020-06-12 | 中国科学院大连化学物理研究所 | Multi-organ chip based on microfluidic technology and application thereof |
| CN111269831A (en) * | 2018-12-05 | 2020-06-12 | 中国科学院大连化学物理研究所 | Transparent multilayer film-sandwiched micro-fluidic chip and preparation method and application thereof |
| CN111269833A (en) * | 2018-12-05 | 2020-06-12 | 中国科学院大连化学物理研究所 | Human pancreatic island organoid model construction method based on organ chip |
| CN109517739A (en) * | 2018-12-27 | 2019-03-26 | 重庆大学 | Double organs are coupled three-layer laminated integrated chip |
| CN110117524A (en) * | 2019-04-17 | 2019-08-13 | 华南理工大学 | A kind of micro flow control chip device of hydrodynamic shear induction tumour cell cross-film migration |
| CN110106081B (en) * | 2019-05-13 | 2022-10-14 | 大连医科大学附属第一医院 | Micro-fluidic chip for constructing brain function unit model and construction method |
| CN110106081A (en) * | 2019-05-13 | 2019-08-09 | 大连医科大学附属第一医院 | For constructing the micro-fluidic chip and construction method of brain function model of element |
| CN110408538A (en) * | 2019-07-18 | 2019-11-05 | 合肥学院 | A kind of liver chip of more lobuli hepatis integrated morphologies |
| CN110408538B (en) * | 2019-07-18 | 2024-01-26 | 安徽骆华生物科技有限公司 | Liver chip of many liver lobule integrated structure |
| CN110643502A (en) * | 2019-10-27 | 2020-01-03 | 苏州济研生物医药科技有限公司 | Single-cell microfluidic detection chip and preparation method and application method thereof |
| CN110669670B (en) * | 2019-11-14 | 2022-06-03 | 大连理工大学 | Human body micro-ecosystem chip and use method thereof |
| CN110669670A (en) * | 2019-11-14 | 2020-01-10 | 大连理工大学 | Human body micro-ecosystem chip and use method thereof |
| CN112553076A (en) * | 2020-12-15 | 2021-03-26 | 东南大学 | Brain organoid in vitro culture chip |
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