CN105754855B - A kind of flow adds two confluent monolayer cells of formula to co-culture organ chip - Google Patents
A kind of flow adds two confluent monolayer cells of formula to co-culture organ chip Download PDFInfo
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- CN105754855B CN105754855B CN201610223671.3A CN201610223671A CN105754855B CN 105754855 B CN105754855 B CN 105754855B CN 201610223671 A CN201610223671 A CN 201610223671A CN 105754855 B CN105754855 B CN 105754855B
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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Abstract
The invention discloses a kind of streams plus two confluent monolayer cells of formula to co-culture organ chip, including chip slot, perforated membrane, jacket layer and head cover, chip slot includes cell culture area of lower layer and three equally distributed choked flow fence, the external distribution of chip slot is equipped with culture supernatants import, upper layer reagent entry port, lower layer's culture solution import, lower layer's reagent entry port, upper layer outlet and lower layer outlet, the front of jacket layer includes upper cell cultivation region and three equally distributed U-shaped choked flow fence, and the back side of jacket layer is equipped with set lamellar spacing and three fixed fences.The present invention has two contactless confluent monolayer cells and co-cultures ability, the contactless information interchange of cell of upper layer and lower layer cultivating system is realized especially by the design of perforated membrane, cell growth status and drug toxicity or pharmacological activity can be detected with real-time online, have the characteristics that simple, portable, economical, efficiently and accurately, the interaction relationship and drug screening and toxicity assessment of a variety of organs and tissue can be generalized to.
Description
Technical field
The present invention relates to a kind of culture organ chip, specifically a kind of stream plus two confluent monolayer cells of formula co-culture organ chip.
Background technology
Cell biology, life science and relevant medical research field are currently carried out under space microgravity and revolving conditions
In, cytologic experiment is mostly the cell culture container using Tissue Culture Flask or specific hard material, and this structure makes own
Cell sample can only use single culture and experiment model, cannot carry out individualized experiments management, it is also difficult to realize multiple organ kind
The interactive research of class cell.Coculture techniques are the cultivating systems for the organ interaction for being more closely similar to vivo environment,
To enable iuntercellular to communicate with each other information, growing multiplication of mutually promoting, and in Cell differentiation inducing activity, maintain cell function and work
Power, regulating cell proliferation, improves and influences each other in metabolite yield and promote and play a positive role.Wherein Co-culture
Be in direct contact between variety classes cell in system is difficult to carry out subsequent single kind cell analysis.Indirect co-culture system makes
Different types of cell shares same cultivating system and is not directly contacted with, and it is porous to mostly use Transwell for routine experiment at present
Plate studies cell migration and interaction, but can not be applied under space microgravity and revolving conditions.Therefore space application is thin
Born of the same parents' training mode is to good cell culture environment, such as device materials gas permeability, biocompatibility etc., experimental implementation it is automatic
Change or simplification, more stringent requirements are proposed for cell long-period culture and Visual retrieval etc..Therefore, in order to adapt to space microgravity
With revolution system and the co-culture system of more histoorgan interaction relationships, it is highly desirable structure and is realized by perforated membrane
The two confluent monolayer cells co-culture system organ chips that contactless cellular informatics exchange.
Invention content
The purpose of the present invention is to provide a kind of simple and convenient, efficiently and accurately streams, and two confluent monolayer cells of formula to be added to co-culture organ core
Piece, to solve the problems mentioned in the above background technology.
To achieve the above object, the present invention provides the following technical solutions：
A kind of flow adds two confluent monolayer cells of formula to co-culture organ chip, including chip slot, perforated membrane, jacket layer and head cover, the top
Lid is mounted on the top of chip slot by bonded seal, and the perforated membrane and jacket layer are sequentially arranged at chip slot and top from top to bottom
The space interior of composition is covered, the chip slot includes cell culture area of lower layer and three equally distributed choked flow fence, chip slot
External distribution culture supernatants import, upper layer reagent entry port, lower layer's culture solution import, lower layer's reagent entry port, upper layer are installed
The front of outlet and lower layer outlet, the jacket layer includes upper cell cultivation region and three equally distributed U-shaped choked flow fence, set
The back side of layer is equipped with set lamellar spacing and three fixed fences, the inside of the U-shaped choked flow fence and jacket layer surrounding are equipped with length
The through channel of bar shaped, the perforated membrane are equipped with multiple through-holes.
As a further solution of the present invention：The aperture of the perforated membrane is 0.4 μm, 1 μm, 3 μm or 5 μm.
As further scheme of the invention：The material of the perforated membrane is PDMS membrane, Transwell
Transparent polyester film or clear polycarbonate film.
Compared with prior art, the beneficial effects of the invention are as follows：
1. the present invention is with easily parallel two or more organ cell types of implantation, two layers of cultivation region can carry out
Monolayer adherence culture or three dimensional gel stroma cell culture, and efficiently separated out by perforated membrane, by culture medium in microchannel
Between interpenetrate, carry out intercellular exchange and influence each other, can be as the interaction of research a variety of organs and tissue
The good platform of relationship and drug screening and toxicity assessment；
2. cell culture fluid may be implemented in the present invention and the stream of other cell experiment reagents adds formula to supply, it can be achieved that when long
Between cell culture update added with multiple cell experiment reagent, make to carry out for a long time cell automatically culture and cytologic experiment as can
Energy；
3. the present invention has independent monolayer cell culture circuit, the cell sample in each cell culture area carries out different
Personalized culture or experiment, quantitative liquid sample can be automatically extracted at the time point of feature according to requirement of experiment, and can be right
Cell sample carries out online image viewing in due course, and cell cultivation process is made to visualize.
Description of the drawings
Fig. 1 is the structural diagram of the present invention.
Fig. 2 is the structural schematic diagram of core film trap and head cover of the present invention.
Fig. 3 is the structural schematic diagram at the jacket layer back side in the present invention.
Fig. 4 is the positive structural schematic diagram of jacket layer in the present invention.
Specific implementation mode
The technical solution of this patent is described in more detail With reference to embodiment.
- 4 are please referred to Fig.1, a kind of flow adds two confluent monolayer cells of formula to co-culture organ chip, including chip slot 1, perforated membrane 2, jacket layer
3 and head cover 4, the head cover 4 is mounted on the top of chip slot 1 by bonded seal, and the perforated membrane 2 and jacket layer 3 are from top to bottom
It is sequentially arranged at the space interior that chip slot 1 and head cover 4 form, the chip slot 1 includes that cell culture area of lower layer 5 and three are equal
The external distribution of the choked flow fence 6 of even distribution, chip slot 1 is equipped with culture supernatants import 7, upper layer reagent entry port 8, lower layer's training
The front of nutrient solution import 9, lower layer's reagent entry port 10, upper layer outlet 11 and lower layer outlet 12, the jacket layer 3 includes upper cell training
Area 13 and three equally distributed U-shaped choked flow fence 14 are supported, the back side of jacket layer 4 is equipped with set lamellar spacing 15 and three fixed fences
16, the inside of the U-shaped choked flow fence 14 and 3 surrounding of jacket layer are equipped with the through channel 17 of strip, on the perforated membrane 2
Equipped with multiple through-holes 18, the aperture of the perforated membrane 2 is 0.4 μm, 1 μm, 3 μm or 5 μm, and the material of the perforated membrane 2 is poly- two
Methylsiloxane film, Transwell transparent polyester films or clear polycarbonate film.
The stream plus two confluent monolayer cells of formula co-culture organ chip when in use, first by the 1 of the merging chip slot of perforated membrane 2
The top of choked flow fence 6, combines closely with edge, so that the through-hole 18 that levels only have perforated membrane 2 is penetrated through, by jacket layer 3
It is face-up to be placed in 2 top of perforated membrane and be closely bonded together, finally head cover 4 and chip slot 1 are bonded, form closing
Two layers co-cultivation organ chip.When cell seeding, first the three-dimensional of the culture solution of the cell concentration of setting or load cells is coagulated
The upper cell cultivation region 5 and lower layer cultivation region 13 of body fluid injection chip before glue, after cell adherent growth or gel forming,
Chip space is being filled up with culture medium.Addition culture supernatants volume there was not U-shaped choked flow fence 14, and culture solution will pass through perforation
Channel 17 is the set lamellar spacing 15 into jacket layer 3, and perforated membrane 2 is completely covered so that bilevel cell culture fluid system
Sufficient mass transfer conversion is carried out by perforated membrane 2, to form Indirect co-culture system.Chip is placed in incubator, you can
Carry out the automatic culture of cell and test, and can be by importing and exporting needed for the realization quantitative addition into tissue culture plate on time or export
Reagent or culture solution, and can be online detection cell growth state, and carry out relevant characterization.
For the present invention with easily parallel two or more organ cell types of implantation, two layers of cultivation region can carry out list
Layer adhere-wall culture or three dimensional gel stroma cell culture, and efficiently separated out by perforated membrane 2, by culture medium between microchannel
Interpenetrate, carry out intercellular exchange and influence each other, the interaction of a variety of organs of research and tissue pass can be used as
The good platform of system and drug screening and toxicity assessment；Cell culture fluid and other cell experiment reagents may be implemented in the present invention
Stream add formula to supply, it can be achieved that long-time cell culture update is added with multiple cell experiment reagent, make to carry out cell for a long time certainly
Dynamic culture is possibly realized with cytologic experiment；The present invention has an independent monolayer cell culture circuit, in each cell culture area
Cell sample carry out different personalized cultures or experiment, can be automatically extracted at the time point of feature according to requirement of experiment quantitative
Liquid sample, and can to cell sample carry out in due course online image viewing, so that cell cultivation process is visualized.
The present invention has two contactless confluent monolayer cells and co-cultures ability, is realized especially by the design of perforated membrane 2
The contactless information interchange of cell of lower two layers of cultivating system both has stream plus formula cell culture fluid and reagent update, realizes thin
The long-term cultivation of born of the same parents；Personalized culture can be carried out in different cell culture areas again, and cell growth status can be observed online in due course,
Cell growth status and drug toxicity or pharmacological activity can be detected with real-time online, had simple, portable, economical, efficient and accurate
True feature is suitable for the cell training of interaction relationship between research organ and tissue under the conditions of space microgravity or revolution system
Device is supported, and the interaction relationship and drug screening and toxicity assessment of a variety of organs and tissue can be generalized to.
The better embodiment of this patent is explained in detail above, but this patent is not limited to above-mentioned embodiment party
Formula, one skilled in the relevant art within the scope of knowledge, can also be under the premise of not departing from this patent objective
Various changes can be made.
Claims (2)
1. a kind of stream plus two confluent monolayer cells of formula co-culture organ chip, which is characterized in that including chip slot (1), perforated membrane (2), set
Layer (3) and head cover (4), the head cover (4) are mounted on the top of chip slot (1), the perforated membrane (2) and set by bonded seal
Layer (3) is sequentially arranged at the space interior of chip slot (1) and head cover (4) composition from top to bottom, and the chip slot (1) includes lower layer
The external distribution of cell culture area (5) and three equally distributed choked flow fence (6), chip slot (1) is equipped with culture supernatants
Import (7), upper layer reagent entry port (8), lower layer's culture solution import (9), lower layer's reagent entry port (10), upper layer outlet (11) and lower layer
It exports (12), the front of the jacket layer (3) includes upper cell cultivation region (13) and three equally distributed U-shaped choked flow fence
(14), the back side of jacket layer (4) is equipped with set lamellar spacing (15) and three fixed fences (16), the U-shaped choked flow fence (14) it is interior
Portion and jacket layer (3) surrounding are equipped with the through channel (17) of strip, and the perforated membrane (2) is equipped with multiple through-holes (18)；
The material of the perforated membrane (2) is PDMS membrane, Transwell transparent polyester films or clear polycarbonate film；
The stream plus two confluent monolayer cells of formula co-culture organ chip when in use, and perforated membrane (2) is placed in (1) of chip slot first
The top of choked flow fence (6), combines closely with edge, so that the through-hole (18) that levels only have perforated membrane (2) is penetrated through, will cover
The face-up of layer (3) is placed in above perforated membrane (2) and is closely bonded together, and finally carries out head cover (4) and chip slot (1)
Bonding forms closed two layers of co-cultivation organ chip；When cell seeding, first by the culture solution of the cell concentration of setting or load
The upper cell cultivation region (5) and lower layer cultivation region (13) of body fluid injection chip, wait for the adherent life of cell before the three dimensional gel of cell
After long or gel forming, chip space is being filled up with culture medium；Addition culture supernatants volume there was not U-shaped choked flow fence
(14), culture solution will enter the set lamellar spacing (15) of jacket layer (3) by through channel (17), and perforated membrane is completely covered
(2) so that bilevel cell culture fluid system carries out sufficient mass transfer conversion by perforated membrane (2), indirect to be formed
Co-culture system；Chip is placed in incubator, cell culture and experiment automatically are carried out, by import and export realize it is on time quantitative to
Reagent or culture solution needed for being added or exporting in tissue culture plate, and on-line checking cell growth state, carry out relevant characterization.
2. stream according to claim 1 plus two confluent monolayer cells of formula co-culture organ chip, which is characterized in that the perforated membrane
(2) aperture is 0.4 μm, 1 μm, 3 μm or 5 μm.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610223671.3A CN105754855B (en) | 2016-04-12 | 2016-04-12 | A kind of flow adds two confluent monolayer cells of formula to co-culture organ chip |
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| CN201610223671.3A CN105754855B (en) | 2016-04-12 | 2016-04-12 | A kind of flow adds two confluent monolayer cells of formula to co-culture organ chip |
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| CN105754855A CN105754855A (en) | 2016-07-13 |
| CN105754855B true CN105754855B (en) | 2018-08-21 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106544270B (en) * | 2016-12-06 | 2019-03-26 | 北京理工大学 | A kind of micro-fluidic chip and its cell culture processes co-cultured for cell |
| CN107502547A (en) * | 2017-09-25 | 2017-12-22 | 中科芯瑞(苏州)生物科技有限公司 | A kind of micro-fluidic chip for realizing various kinds of cell co-cultivation and its application |
| CN108384719B (en) * | 2018-04-26 | 2023-11-17 | 中国烟草总公司郑州烟草研究院 | cell co-culture device |
| CN112300929B (en) * | 2019-07-31 | 2022-12-02 | 上海新微技术研发中心有限公司 | Microfluidic experiment plate and double-sided cell culture method |
| CN111394248A (en) * | 2020-04-23 | 2020-07-10 | 西安交通大学口腔医院 | Cell culture experimental device |
| CN113388517B (en) * | 2021-06-08 | 2022-10-25 | 北京理工大学 | Biological culture micro-fluidic chip suitable for assembling microgravity gyroscope and cell culture method thereof |
| CN113388516B (en) * | 2021-06-25 | 2022-03-11 | 上海睿钰生物科技有限公司 | Culture device |
| CN114015568B (en) * | 2021-10-08 | 2024-02-23 | 北京龙迈达斯科技开发有限公司 | Organoid chip and preparation method thereof |
| CN114317272B (en) * | 2021-12-07 | 2024-01-12 | 上海前瞻创新研究院有限公司 | Culture device for multicellular co-culture and cell culture method |
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| US9023642B2 (en) * | 2006-07-07 | 2015-05-05 | The University Of Houston System | Method and apparatus for a miniature bioreactor system for long-term cell culture |
| EP2634251A1 (en) * | 2012-02-29 | 2013-09-04 | Technische Universität Berlin | 3D in vitro bi-phasic cartilage-bone construct |
| CN103981096B (en) * | 2014-05-27 | 2015-11-18 | 东南大学 | A kind of two-layer cell culture system organ chip and preparation method thereof |
| CN103981085B (en) * | 2014-05-27 | 2016-01-06 | 东南大学 | A kind of from establishing concentration gradient drug screening organ chip and preparation method thereof |
| CN203976812U (en) * | 2014-08-07 | 2014-12-03 | 中国航天员科研训练中心 | The controlled locked cell culture chamber of the applied stream in space |
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