CN105754855A - Fed-batch type two-layer cell co-culture chip - Google Patents
Fed-batch type two-layer cell co-culture chip Download PDFInfo
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- CN105754855A CN105754855A CN201610223671.3A CN201610223671A CN105754855A CN 105754855 A CN105754855 A CN 105754855A CN 201610223671 A CN201610223671 A CN 201610223671A CN 105754855 A CN105754855 A CN 105754855A
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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
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis
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Abstract
The invention discloses a fed-batch type two-layer cell co-culture chip which comprises a chip slot, a porous membrane, a sleeve layer and a top cover, wherein the chip slot comprises a lower cell culture area and three uniformly distributed flow choking barriers; an upper culture liquid inlet, an upper reagent inlet, a lower culture liquid inlet, a lower reagent inlet, an upper outlet and a lower outlet are formed in the outer part of the chip slot; an upper cell culture area and three uniformly distributed U-shaped flow choking barriers are arranged on the front side of the sleeve layer; a sleeve layer gap and three fixed barriers are arranged on the back side of the sleeve layer. The fed-batch type two-layer cell co-culture chip is capable of co-culturing two layers of cells, particularly due to design of the porous membrane, cell non-contact type information communication of upper and lower layers of culture systems can be achieved, the growth situation of cells and drug toxicity or pharmacology activity can be detected at real time on line, and the fed-batch type two-layer cell co-culture chip has the characteristics of simplicity, portability, economy, efficiency and accuracy, and can be popularized to mutual action relationship of multiple organs and tissues, medicine screening and toxicity evaluation.
Description
Technical field
The present invention relates to a kind of cultivation organ chip, specifically a kind of stream adds formula two-layer co-culture of cells organ chip.
Background technology
Currently carry out in cytobiology, life sciences and relevant medical research field under space microgravity and revolving conditions, mostly cytologic experiment is to adopt the cell culture container of Tissue Culture Flask or specific hard material, this structure makes all cells sample can only adopt single cultivation and experiment model, individualized experiments management can not be carried out, it is also difficult to realize the interactive research of multiple organ cell types.Coculture techniques is to be more closely similar to the cultivating system that the organ of internal milieu interacts, the information so that iuntercellular can be communicated with each other, mutually promote growing multiplication, and at Cell differentiation inducing activity, maintain cell function and vigor, regulating cell is bred, and improves and influences each other in metabolite yield and promote and play a positive role.Wherein Co-culture system directly contacts between variety classes cell and be difficult to carry out follow-up single kind cell analysis.Indirect co-culture system makes different types of cell share same cultivating system and be not directly contacted with, and current normal experiment many employings Transwell porous plate studies cell migration and interaction, but cannot be applied under space microgravity and revolving conditions.Therefore application cell training mode in space is to good cell culture environment, such as device materials breathability, biocompatibility etc., the automatization of experimental implementation or simplification, and cell long-period is cultivated and Visual retrieval etc. is had higher requirement.Therefore, in order to adapt to space microgravity and revolution system, and the co-culture system of many histoorgans interaction relationship, it is highly desirable to build the two-layer co-culture of cells system organ chip being realized the exchange of contactless cellular informatics by perforated membrane.
Summary of the invention
It is an object of the invention to provide a kind of stream simple and convenient, efficiently and accurately and add formula two-layer co-culture of cells organ chip, with the problem solving to propose in above-mentioned background technology.
For achieving the above object, the present invention provides following technical scheme:
A kind of stream adds formula two-layer co-culture of cells organ chip, including chip slot, perforated membrane, jacket layer and top cover, described top cover is arranged on the top of chip slot by bonded seal, described perforated membrane and jacket layer are sequentially arranged at chip slot and the interior volume of top cover composition from top to bottom, described chip slot includes lower confluent monolayer cells cultivation region and three equally distributed choked flow fence, the outside distribution of chip slot is provided with culture supernatants import, upper strata reagent entry port, lower floor's culture fluid import, lower floor's reagent entry port, upper strata outlet and lower floor's outlet, the front of described jacket layer includes confluent monolayer cells cultivation region and three equally distributed U-shaped choked flow fence, the back side of jacket layer is provided with set lamellar spacing and three fixed fences, the inside of described U-shaped choked flow fence and jacket layer surrounding are equipped with the through channel of strip, described perforated membrane is provided with multiple through hole.
As the further scheme of the present invention: the aperture of described perforated membrane is 0.4 μm, 1 μm, 3 μm or 5 μm.
As the present invention further scheme: the material of described perforated membrane is PDMS membrane, Transwell transparent polyester film or clear polycarbonate film.
Compared with prior art, the invention has the beneficial effects as follows:
1. the present invention is with two or more organ cell types of parallel implantation easily, two-layer cultivation region all can carry out monolayer adherence cultivation or three dimensional gel stromal cell is cultivated, and efficiently separated out by perforated membrane, by culture medium interpenetrating between microchannel, carry out intercellular exchange and influence each other, it is possible to as the good platform of the interaction relationship and drug screening and toxicity assessment of studying multiple organ and tissue；
2. the present invention can realize the stream of cell culture fluid and other cell experiment reagent and adds formula supply, it may be achieved long-time cell is cultivated to update and added with repeatedly cell experiment reagent, makes to carry out for a long time cell and automatically cultivates and be possibly realized with cytologic experiment；
3. the present invention possesses independent monolayer cell culture loop, cell sample in each cell cultivation region carries out different personalized cultivations or experiment, quantitative liquid sample can be automatically extracted at the time point of feature according to requirement of experiment, and cell sample can be carried out online image viewing in good time, make cell cultivation process visualize.
Accompanying drawing explanation
Fig. 1 is the structural representation of the present invention.
Fig. 2 is the structural representation of SMIS film trap of the present invention and top cover.
Fig. 3 is the structural representation at the jacket layer back side in the present invention.
Fig. 4 is the structural representation in jacket layer front in the present invention.
Detailed description of the invention
Below in conjunction with detailed description of the invention, the technical scheme of this patent is described in more detail.
nullRefer to Fig. 1-4，A kind of stream adds formula two-layer co-culture of cells organ chip，Including chip slot 1、Perforated membrane 2、Jacket layer 3 and top cover 4，Described top cover 4 is arranged on the top of chip slot 1 by bonded seal，Described perforated membrane 2 and jacket layer 3 are sequentially arranged at chip slot 1 and the interior volume of top cover 4 composition from top to bottom，Described chip slot 1 includes lower confluent monolayer cells cultivation region 5 and three equally distributed choked flow fence 6，The outside distribution of chip slot 1 is provided with culture supernatants import 7、Upper strata reagent entry port 8、Lower floor's culture fluid import 9、Lower floor's reagent entry port 10、Upper strata outlet 11 and lower floor's outlet 12，The front of described jacket layer 3 includes confluent monolayer cells cultivation region 13 and three equally distributed U-shaped choked flow fence 14，The back side of jacket layer 4 is provided with set lamellar spacing 15 and three fixed fences 16，The inside of described U-shaped choked flow fence 14 and jacket layer 3 surrounding are equipped with the through channel 17 of strip，Described perforated membrane 2 is provided with multiple through hole 18，The aperture of described perforated membrane 2 is 0.4 μm、1μm、3 μm or 5 μm，The material of described perforated membrane 2 is PDMS membrane、Transwell transparent polyester film or clear polycarbonate film.
Described stream adds formula two-layer co-culture of cells organ chip in use, first perforated membrane 2 is inserted chip slot 1 the top of choked flow fence 6, be combined with edges close, the through hole 18 that levels only has perforated membrane 2 is made to carry out through, facing up of jacket layer 3 is placed in above perforated membrane 2 and is closely bonded together, finally top cover 4 and chip slot 1 are bonded, form the two-layer closed and co-culture organ chip.During cell seeding, first body fluid before the culture fluid of cell concentration set or the three dimensional gel of load cells is injected upper confluent monolayer cells cultivation region 5 and the lower floor cultivation region 13 of chip, after treating cell attachment growth or gel forming, filling up chip space by culture medium.Add culture supernatants volume and do not have U-shaped choked flow fence 14, culture fluid will pass through through channel 17 and namely enter the set lamellar spacing 15 of jacket layer 3, and perforated membrane 2 is completely covered, bilevel cell culture fluid system is made to carry out sufficient mass transfer conversion by perforated membrane 2, thus forming Indirect co-culture system.Chip is placed in incubator, cell can be carried out and automatically cultivate and test, and the reagent needed for import and export realization quantitatively adds in Tissue Culture Plate or derives on time or culture fluid can be passed through, it is possible to online detection cell growth state, and carry out relevant characterization.
The present invention is with two or more organ cell types of parallel implantation easily, two-layer cultivation region all can carry out monolayer adherence cultivation or three dimensional gel stromal cell is cultivated, and efficiently separated out by perforated membrane 2, by culture medium interpenetrating between microchannel, carry out intercellular exchange and influence each other, it is possible to as the good platform of the interaction relationship and drug screening and toxicity assessment of studying multiple organ and tissue；The present invention can realize the stream of cell culture fluid and other cell experiment reagent and add formula supply, it may be achieved long-time cell is cultivated to update and added with repeatedly cell experiment reagent, makes to carry out for a long time cell and automatically cultivates and be possibly realized with cytologic experiment；The present invention possesses independent monolayer cell culture loop, cell sample in each cell cultivation region carries out different personalized cultivations or experiment, quantitative liquid sample can be automatically extracted at the time point of feature according to requirement of experiment, and cell sample can be carried out online image viewing in good time, make cell cultivation process visualize.
The present invention possesses contactless two-layer co-culture of cells ability, realizes the contactless communication for information of cell of upper and lower two-layer cultivating system especially by the design of perforated membrane 2, had both possessed stream and has added formula cell culture fluid and reagent updates, it is achieved the long-term cultivation of cell；Personalized cultivation can be carried out again in different cell cultivation regions, and can online observation of cell growing state in good time, cell growth status and drug toxicity or pharmacologically active can be detected by real-time online, there is feature simple, portable, economic, efficiently and accurately, suitable in the cell culture system of research organ and tissue interphase interaction relation under space microgravity or revolution system condition, and the interaction relationship of multiple organ and tissue and drug screening and toxicity assessment can be generalized to.
Above the better embodiment of this patent is explained in detail, but this patent is not limited to above-mentioned embodiment, in the ken that one skilled in the relevant art possesses, it is also possible under the premise without departing from this patent objective, make various change.
Claims (3)
- null1. a stream adds formula two-layer co-culture of cells organ chip，It is characterized in that，Including chip slot (1)、Perforated membrane (2)、Jacket layer (3) and top cover (4)，Described top cover (4) is arranged on the top of chip slot (1) by bonded seal，The interior volume that described perforated membrane (2) and jacket layer (3) are sequentially arranged at chip slot (1) from top to bottom and top cover (4) forms，Described chip slot (1) includes lower confluent monolayer cells cultivation region (5) and three equally distributed choked flow fence (6)，The outside distribution of chip slot (1) is provided with culture supernatants import (7)、Upper strata reagent entry port (8)、Lower floor's culture fluid import (9)、Lower floor's reagent entry port (10)、Upper strata outlet (11) and lower floor's outlet (12)，The front of described jacket layer (3) includes confluent monolayer cells cultivation region (13) and three equally distributed U-shapeds choked flow fence (14)，The back side of jacket layer (4) is provided with set lamellar spacing (15) and three fixed fences (16)，The inside of described U-shaped choked flow fence (14) and jacket layer (3) surrounding are equipped with the through channel (17) of strip，Described perforated membrane (2) is provided with multiple through hole (18).
- 2. stream according to claim 1 adds formula two-layer co-culture of cells organ chip, it is characterised in that the aperture of described perforated membrane (2) is 0.4 μm, 1 μm, 3 μm or 5 μm.
- 3. stream according to claim 1 adds formula two-layer co-culture of cells organ chip, it is characterised in that the material of described perforated membrane (2) is PDMS membrane, Transwell transparent polyester film or clear polycarbonate film.
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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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Cited By (9)
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| CN106544270A (en) * | 2016-12-06 | 2017-03-29 | 北京理工大学 | A kind of micro-fluidic chip and its cell culture processes for co-culture of cells |
| 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 |
| CN108384719A (en) * | 2018-04-26 | 2018-08-10 | 中国烟草总公司郑州烟草研究院 | Cell co-culture device |
| CN111394248A (en) * | 2020-04-23 | 2020-07-10 | 西安交通大学口腔医院 | Cell culture experimental device |
| CN112300929A (en) * | 2019-07-31 | 2021-02-02 | 上海新微技术研发中心有限公司 | Microfluidic experimental plate and double-sided cell culture method |
| CN113388517A (en) * | 2021-06-08 | 2021-09-14 | 北京理工大学 | Biological culture micro-fluidic chip suitable for assembling microgravity gyroscope and cell culture method thereof |
| CN113388516A (en) * | 2021-06-25 | 2021-09-14 | 上海睿钰生物科技有限公司 | Culture device |
| CN114015568A (en) * | 2021-10-08 | 2022-02-08 | 北京龙迈达斯科技开发有限公司 | Organoid chip and preparation method thereof |
| CN114317272A (en) * | 2021-12-07 | 2022-04-12 | 上海前瞻创新研究院有限公司 | Multi-cell co-culture device and cell culture method |
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Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN106544270A (en) * | 2016-12-06 | 2017-03-29 | 北京理工大学 | A kind of micro-fluidic chip and its cell culture processes for co-culture of cells |
| 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 |
| CN108384719A (en) * | 2018-04-26 | 2018-08-10 | 中国烟草总公司郑州烟草研究院 | Cell co-culture device |
| CN112300929A (en) * | 2019-07-31 | 2021-02-02 | 上海新微技术研发中心有限公司 | Microfluidic experimental 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 |
| CN113388517A (en) * | 2021-06-08 | 2021-09-14 | 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 |
| CN113388516A (en) * | 2021-06-25 | 2021-09-14 | 上海睿钰生物科技有限公司 | Culture device |
| CN114015568A (en) * | 2021-10-08 | 2022-02-08 | 北京龙迈达斯科技开发有限公司 | Organoid chip and preparation method thereof |
| 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 |
| CN114317272A (en) * | 2021-12-07 | 2022-04-12 | 上海前瞻创新研究院有限公司 | Multi-cell co-culture device and cell culture method |
| 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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