CN108641931A - A kind of digitlization microarray organ chip and its application - Google Patents
A kind of digitlization microarray organ chip and its application Download PDFInfo
- Publication number
- CN108641931A CN108641931A CN201810298289.8A CN201810298289A CN108641931A CN 108641931 A CN108641931 A CN 108641931A CN 201810298289 A CN201810298289 A CN 201810298289A CN 108641931 A CN108641931 A CN 108641931A
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- China
- Prior art keywords
- microarray
- digitlization
- organ chip
- bottom plate
- upper chamber
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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
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/16—Particles; Beads; Granular material; Encapsulation
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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
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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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
Abstract
The present invention provides a kind of digitlization microarray organ chip, including top plate, PET film and bottom plate；Upper chamber is equipped in top plate, lower chambers are equipped in bottom plate, upper chamber is consistent with the shape of lower chambers, it is separated by PET film between upper chamber and lower chambers, the bottom chamber of bottom plate is arranged with the microarray hole for microballoon culture and digital measuring, input channel and output channel are connected on top plate, input channel and output channel are connected to upper chamber respectively.The present invention has the characteristics that bionical, high-throughput, cheap, has preferable foreground in terms of drug screening, disease modeling and the clinical applications such as cytotoxicity test.
Description
Technical field
The present invention relates to biotechnologies, and in particular to a kind of digitlization microarray organ core based on microflow control technique
Piece and its application.
Background technology
" organ chip " technology be in microfluidic devices by way of continuous pouring culture living cells simulating group
Knit the physiological function with organ.It can be provided for cell growth in analogue body physical environment (hydrodynamic shear, cyclic strain and
Mechanical compression etc.), so that it is more really represented cell and organ.Liquid flowing can be controlled in organ chip, to enhance
The differentiation of many cell types and the long-term holding of function.Existing organ chip can utilize microflow control technique, microfabrication
Carry out three-dimensional microenvironment complicated in analogue body with biomaterial, such as cultivates the micro- blood of people in 3D institutional frameworks using microfluidic device
Endothelial cell simulates the microchannel of lymphatic vessel and blood vessel.The tumor phenotype observed in vivo can also be simulated, i.e.,
By necrotic cores, the gradient shape knot of internal stationary cell ring and external proliferative cell zone composition.
Digital assay is a kind of current accurate analysis system of high throughput, can be in microstructure to analytic unit point
It cuts and independently analyzes, there is the features such as high collimation, high duplication, can be used for improving the accurate analysis level of organ chip.Currently,
Digitlization medicine sorting platform is all based on 384 and 1536 orifice plates, but this method is limited to orifice plate number in precision, and can not
Realize 3D dynamic cultivations.Simultaneously, it is difficult to be further reduced the consumption of reagent, amount of pixels is limited to orifice plate number, is difficult to carry out liquid
The accurate distribution of body, and influenced by liquid evaporation.And the digital assay based on microflow control technique, there are mainly two types of moulds
Formula：Microarray format and droplet pattern.Digital assay based on droplet pattern, it is difficult to produced by removing in cell growth process
Toxic metabolic substance, and addition nutriment, greatly limit research application of the droplet analysis platform in organ chip.Though
So it is provided with the blank of digital assay with the organ chip technology that micropore is basic microarray, but microarray organ core at present
Piece is all to be retained in the chip to cell, agglomerating culture, all suffers from the lower defect of flux.
Therefore, develop a kind of digitlization microarray organ chip, by carrying out the culture of 3D microballoons and inspection in microporous matrix
It surveys, such 3D microballoons generate 3D sphere gel stents, certain hole can be generated after freeze-drying by droplet generating principle, high throughput,
It can be cultivated after freeze-drying holder adherent cell.Such method is the high-throughput preparation outside digitlization organ chip, is different from
The characteristics of existing organ chip is to be retained in the chip to cell, agglomerating culture.The digitlization microarray organ chip
It is to be cultivated and detected after microballoon containing cell is directly injected into.The digitized microarray organ chip overcomes at present
There is the lower defect of the flux of organ chip, realizes high-throughput digital assay.The digitlization organ chip platform has
Precision is high, reproducible feature, can be in drug screening, and disease modeling, medical compounds optimization and cytotoxicity test need
Aspect is asked to play a role.
Invention content
In view of this, the technical problem to be solved in the present invention is to overcome the deficiencies of existing technologies, a kind of number is provided
Change microarray organ chip, including top plate, PET film and bottom plate；The top plate includes liquid input, output channel, the top
Plate, PET film form upper chamber and form lower chambers, the upper and lower chamber quilt for adding cell culture fluid, the PET film and bottom plate
One layer of PET film separates.The bottom plate is arranged with microarray hole, and the microarray hole is used for microballoon culture and digital measuring.
In some embodiments, the top plate, PET film, bottom plate are sequentially connected by ionic bond；The top plate offers
Liquid injection hole, drainage hole and note ball, gas vent, are communicated with liquid injection pipe and drain pipe, the note at the liquid injection hole and drainage hole
Note bulb and exhaust pipe are communicated at ball and gas vent.It is cell culture fluid perfusion chamber that the top plate, which has upper chamber,.
In some embodiments, PET film is equipped between the upper and lower chamber, aperture is 3~10 μm.
In some embodiments, the lower chambers are located at immediately below the film, cross-sectional area 1-3cm2。
In some embodiments, on the bottom plate it is micropore, number is 200~10,000, and aperture is 100~300 μ
M, thickness are 100~200 μm.
In some embodiments, the epicoele chamber is U-shaped structure.
In some embodiments, the liquid injection hole aperture is 1-2mm；The liquid discharge orifice aperture is 1-2mm；It is described
It is 1-2mm to note ball aperture；The gas vent aperture is 1-2 mm；The liquid injection pipe caliber is 0.5-1mm, length 8-
10cm；The drain pipe caliber is 0.5-1mm, length 8-10cm；The note bulb caliber is 0.5-1mm, length 1-
2cm；The exhaust pipe caliber is 0.5-1mm, length 1-2cm.
In some embodiments, the top plate and bottom plate are PDMS plates, and its length of side is respectively 10-15mm；It is described
The thickness of top plate and bottom plate is respectively 2-5mm.
The present invention also provides a kind of applications of digitlization microarray organ chip, which is characterized in that includes the following steps：
S10：In the bottom plate for the lower chambers that microballoon containing cell is injected into described in claim 4,5,7, clip is used
Closing note ball and gas vent, stand 4 hours；
S20：Cell culture fluid is injected in the upper chamber described in claim 6；
S30：It is connected with peristaltic pump culture to digitizing microarray organ chip in S20 steps, cultivates and after a certain period of time may be used
Carry out microarray analysis.
In some embodiments, when cell or microballoon being injected into digitlization microarray organ chip, sample introduction speed
For 0.5-2mL/min.
A kind of digitlization microarray organ chip advantageous effect provided by the invention is：
Digitlization microarray organ chip be micro-fluid chip, one piece several square centimeters of chips can be to containing difference
The long-term cultivation that the microballoon of cell type is recycled, and a kind of platform of high throughput analysis can be carried out to its cell activity.This
It is that one kind can be used for drug screening, the Important Platform of disease modeling and Study of cytotoxicity.With bionical, high-throughput, cheap etc.
Feature.The digitlization microarray organ chip preferably simulated in vivo environment and can carry out high throughput analysis, possess huge city
Fieldization foreground.
In conclusion the present invention is in view of the problems of the existing technology, using the microarray on organ chip, one is established
Kind high-flux detection method.It the digitlization microarray organ chip and its applies in drug screening, disease modeling and cytotoxicity
Test etc. has preferable application prospect.
Description of the drawings
It should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore it is not construed as to model
The restriction enclosed for those of ordinary skill in the art without creative efforts, can also be according to these
Attached drawing obtains other relevant attached drawings.
Fig. 1 is the structural decomposition diagram of invention.
Fig. 2 is the constitutional diagram of the present invention.
Fig. 3 is the constitutional diagram of the present invention.
Drawing reference numeral explanation：Top plate 1, liquid injection hole 1-1, drainage hole 1-2 note ball 1-3, gas vent 1-4, upper chamber 1-5,
Liquid injection pipe 1-6, drain pipe 1-7 note bulb 1-8, exhaust pipe 1-9, PET film 2-1, bottom plate 3, lower chambers 3-1, micropore 3-1-1.
Specific implementation mode
The embodiment of the present invention is described below in detail, examples of the embodiments are shown in the accompanying drawings, wherein from beginning to end
Same or similar label indicates same or similar element or element with the same or similar functions.Below with reference to attached
The embodiment of figure description is exemplary, it is intended to for explaining the present invention, and is not considered as limiting the invention.
In the description of the present invention, it is to be understood that, term " length ", " width ", "upper", "lower", "front", "rear",
The orientation or positional relationship of the instructions such as "left", "right", "vertical", "horizontal", "top", "bottom" "inner", "outside" is based on attached drawing institute
The orientation or positional relationship shown, is merely for convenience of description of the present invention and simplification of the description, and does not indicate or imply the indicated dress
It sets or element must have a particular orientation, with specific azimuth configuration and operation, therefore should not be understood as the limit to the present invention
System.
In the present invention, it is limited unless there are specific, in the present description, PET film pore diameter range can be adjusted, greatly
Small is 3~10 μm, while it should be understood that as digitlization microarray organ microarray biochip, material can be PDMS, but
It is not limited to PDMS, it can also be for PMMA or other can be carved or hollow out for the material with the big small-bore of correspondence is this patent institute
Protection domain.In addition, term " upper chamber ", " lower chambers " are used for description purposes only, it is not understood to indicate or imply opposite
Importance implicitly indicates indicated technical characteristic.
In the present invention unless specifically defined or limited otherwise, the terms such as term " connected ", " connection ", " fixation " are answered
It is interpreted broadly, for example, it may be being fixedly connected, may be a detachable connection, or is integral；Can be mechanical connection,
It can be electrical connection；It can be directly connected, can also can be indirectly connected through an intermediary the company inside two elements
Logical or two elements interaction relationship.For the ordinary skill in the art, can understand as the case may be
The concrete meaning of above-mentioned term in the present invention.
Embodiment
Referring to Fig. 1, the present invention provides a kind of digitlization microarray organ chips, including top plate, PET film and bottom plate；
The top plate includes liquid input, output channel, and the top plate, PET films form upper chamber and be used to add cell culture fluid, institute
It states PET film and bottom plate forms lower chambers, the upper and lower chamber is separated by one layer of PET film.The bottom plate is arranged with microarray hole, institute
Microarray hole is stated for microballoon culture and digital measuring.
It is above-mentioned, it is to be understood that digitlization microarray organ chip is micro-fluid chip, at one piece several square centimeters
The long-term cultivation that chip can recycle the microballoon containing different cell types, and its cell activity can be carried out high-throughput
A kind of platform of analysis can be used for drug screening, disease modeling and Study of cytotoxicity.The chip has bionical, high-throughput, honest and clean
The features such as valence.The digitlization microarray organ chip preferably simulated in vivo environment and can carry out high throughput analysis, possess huge
Market-oriented foreground.
In conclusion the present invention is in view of the problems of the existing technology, using the microarray on organ chip, one is established
Kind high-flux detection method.It the digitlization microarray organ chip and its applies in drug screening, disease modeling and cytotoxicity
Test etc. has preferable application prospect.
In embodiments of the present invention, the top plate, PET film, bottom plate are sequentially connected by ionic bonding；The top plate opens up
There are liquid injection hole, drainage hole and note ball, gas vent, liquid injection pipe and drain pipe are communicated at the liquid injection hole and drainage hole, it is described
Note bulb and exhaust pipe are communicated at note ball and gas vent.
In embodiments of the present invention, PET film is equipped between the upper and lower chamber, aperture is 3 μm.
In embodiments of the present invention, the upper chamber is U-shaped structure, and cross-sectional area is 1 cm2.The structure can be used for training
Perfusion and the constant flow for supporting base, are conducive to mass exchange and long-term cultivation.
In embodiments of the present invention, the liquid injection hole aperture is 1mm；The liquid discharge orifice aperture is 1mm；The note ball
Hole aperture is 1mm；The gas vent aperture is 1mm；The liquid injection pipe caliber is 1mm, length 10cm；The drain pipe caliber
For 1mm, length 10cm；The note bulb caliber is 1mm, length 1cm；The exhaust pipe caliber is 1mm, length 2cm.
The structure can be used for adding culture medium and microballoon respectively, be conducive to the fixation and culture of microballoon.
In embodiments of the present invention, the top plate and bottom plate are PDMS plates, and its length of side is respectively 1.2cm；The top
The thickness of plate and bottom plate is followed successively by 2mm.
In embodiments of the present invention, on the bottom plate it is micropore, number 10,000, aperture is 100 μm, and thickness is
100μm.Due to the structure design of micropore, when in use, structure of the invention can reach 10,000 flux of unit interval, phase
It commonly uses more in the prior art for less than 100 flux of product, analysis efficiency is greatly improved.
The present invention also provides a kind of applications of digitlization microarray organ chip, include the following steps：
S10：In the bottom plate for the lower chambers that microballoon containing cell is injected into described in claim 4,5,7, clip is used
Closing note ball and gas vent, stand 4 hours；
S20：Cell culture fluid is injected in the upper chamber described in claim 6；
S30：Microarray organ chip will be digitized in S20 steps with peristaltic pump to be connected culture, cultivate and after a certain period of time may be used
Carry out microarray analysis.
It is understood that miniflow syringe pump, autosampler or manual injector can be used in step S10 by collection
Microballoon is injected into the digitlization microarray organ chip 1.
It is understood that after step S30 cell activation assay can be carried out to microarray.
It is to be appreciated that the chip can directly be detected the microballoon in micro array structure, this kind of detection mode
As in situ detection.
It is to be appreciated that under the microballoon in micro array structure on digitlization microarray organ chip is rinsed from chip
Come, then the process being detected detects for ex situ.
In embodiments of the present invention, when microballoon or cell being injected into digitlization microarray organ chip 1, sample introduction speed
Degree is 1mL/min.
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, for the skill of this field
For art personnel, the invention may be variously modified and varied.All within the spirits and principles of the present invention, any made by repair
Change, equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of digitlization microarray organ chip, it is characterised in that：Including top plate, PET film and bottom plate；It is equipped in top plate upper
Chamber is equipped with lower chambers in bottom plate, and upper chamber is consistent with the shape of lower chambers, between upper chamber and lower chambers by PET film every
It opens, the bottom chamber of bottom plate is arranged with the microarray hole for microballoon culture and digital measuring, and input pipe is connected on top plate
Road and output channel, input channel and output channel are connected to upper chamber respectively.
2. a kind of digitlization microarray organ chip as described in claim 1, it is characterised in that：The top plate, PET film, bottom
Plate is sequentially connected by ionic bond, and the aperture of PET film is 3~10 μm.
3. a kind of digitlization microarray organ chip as described in claim 1, it is characterised in that：Input channel includes liquid injection pipe
With note bulb, output channel includes drain pipe and exhaust pipe.
4. a kind of digitlization microarray organ chip as described in claim 1, it is characterised in that：It opens respectively at the both ends of upper chamber
If liquid injection hole and drainage hole, liquid injection hole are connected to liquid injection pipe, drainage hole is connected to drain pipe；Be further opened on top plate note ball and
Gas vent, note ball are connected to note bulb, and gas vent is connected to exhaust pipe, and note ball is connected to liquid injection hole, gas vent and drain
Hole is connected to.
5. a kind of digitlization microarray organ chip as claimed in claim 2, it is characterised in that：The microarray hole, is located at
Below the PET film, shared total cross-sectional area is 1-3cm2。
6. a kind of digitlization microarray organ chip as claimed in claim 4, it is characterised in that：The bottom plate is equipped with micro-
Hole, number are 200~10000, and 100~300 μm of aperture, thickness is 100~200 μm.
7. a kind of digitlization microarray organ chip as claimed in claim 4, it is characterised in that：The upper chamber is U-shaped knot
Structure, the both sides of U-shaped structure are respectively used to that ball and liquid injection hole and gas vent will be noted and drainage hole is respectively communicated with.
8. a kind of digitlization microarray organ chip as claimed in claim 4, it is characterised in that：The aperture of the liquid injection hole is
1-2mm；The aperture of the liquid discharge orifice is 1-2mm；The aperture of the note ball is 1-2mm；The gas vent aperture is 1-
2mm；The liquid injection pipe caliber is 0.5-1mm, length 8-10cm；The drain pipe caliber is 0.5-1mm, length 5-
10cm；The note bulb caliber is 0.5-1mm, length 1-2cm；The exhaust pipe caliber is 0.5-1mm, length 1-2cm.
9. a kind of digitlization microarray organ chip as claimed in claim 2, it is characterised in that：The top plate and bottom plate are
The PDMS plates of square, and its length of side is respectively 10-15mm；The thickness of the top plate and bottom plate is respectively 2-5mm.
10. a kind of application of digitlization microarray organ chip, which is characterized in that include the following steps：
S10：In the bottom plate for the lower chambers that microballoon containing cell is injected into described in claim 4,5,7, closed with clip
Ball and gas vent are noted, stands 4 hours；
S20：Cell culture fluid is injected in the upper chamber described in claim 6；
S30：It is connected with peristaltic pump culture to digitizing microarray organ chip in S20 steps, cultivating can carry out after a certain period of time
Microarray analysis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201810298289.8A CN108641931A (en) | 2018-04-04 | 2018-04-04 | A kind of digitlization microarray organ chip and its application |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810298289.8A CN108641931A (en) | 2018-04-04 | 2018-04-04 | A kind of digitlization microarray organ chip and its application |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109055204A (en) * | 2018-10-19 | 2018-12-21 | 杭州捷诺飞生物科技股份有限公司 | Drug screening organ chip |
| 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 |
| CN112226363A (en) * | 2020-09-14 | 2021-01-15 | 北京大学 | Device and method for culturing high-flux organoid by utilizing microarray deep well |
| WO2021248637A1 (en) * | 2020-06-09 | 2021-12-16 | 苏州大学 | Early embryo simulated fallopian tube environment in-vitro culture chip capable of breaking through growth retardation |
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| CN106399094A (en) * | 2012-04-01 | 2017-02-15 | Emd密理博公司 | Cell culture and gradient migration assay methods and devices |
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| CN208762505U (en) * | 2018-04-04 | 2019-04-19 | 浙江大学 | A kind of digitlization microarray organ chip |
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| CN106399094A (en) * | 2012-04-01 | 2017-02-15 | Emd密理博公司 | Cell culture and gradient migration assay methods and devices |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109055204A (en) * | 2018-10-19 | 2018-12-21 | 杭州捷诺飞生物科技股份有限公司 | Drug screening organ chip |
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| WO2021248637A1 (en) * | 2020-06-09 | 2021-12-16 | 苏州大学 | Early embryo simulated fallopian tube environment in-vitro culture chip capable of breaking through growth retardation |
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| CN112226363B (en) * | 2020-09-14 | 2022-05-17 | 北京大学 | Device and method for culturing high-flux organoid by utilizing microarray deep well |
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