KR20140070206A - Cell culture container and the method of preparation thereof - Google Patents
Cell culture container and the method of preparation thereof Download PDFInfo
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- KR20140070206A KR20140070206A KR1020120138420A KR20120138420A KR20140070206A KR 20140070206 A KR20140070206 A KR 20140070206A KR 1020120138420 A KR1020120138420 A KR 1020120138420A KR 20120138420 A KR20120138420 A KR 20120138420A KR 20140070206 A KR20140070206 A KR 20140070206A
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- South Korea
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- cell culture
- culture container
- carbon nanotube
- carbon nanotubes
- amine group
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- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01B—NON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
- C01B32/00—Carbon; Compounds thereof
- C01B32/15—Nano-sized carbon materials
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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
- C12M23/20—Material Coatings
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- 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
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/0068—General culture methods using substrates
Abstract
TECHNICAL FIELD The present invention relates to a cell culture container and a method of manufacturing the same, and more particularly, to a cell culture container without a cytotoxicity, the surface of which is coated with functional carbon nanotubes, and a method for manufacturing the same. Accordingly, the cell culture container coated with functional carbon nanotubes has an effect of improving cell growth activity without cytotoxicity.
Description
TECHNICAL FIELD The present invention relates to a cell culture container and a method of manufacturing the same, and more particularly, to a cell culture container without a cytotoxicity, the surface of which is coated with functional carbon nanotubes, and a method for manufacturing the same.
Conventionally, cell culture has been known as a technique used only for research purpose. Recently, as cell therapy using cultured cells has been expanded to treat various diseases, interest in cell culture has increased in the medical field.
Artificially cultured cells show various characteristic features during growth, proliferation, and differentiation. Most cells in the cell culture adhere to the bottom of the cell culture vessel and survive the process of growth, proliferation and differentiation. However, some cells form different layers while forming layers of cells, and some of them grow, proliferate, and differentiate while maintaining a floating state in cell culture fluids.
A variety of devices are used in the culture system currently in use, and one of the important factors in this is the cell culture container. The types of cell culture containers currently used in the world include culture dishes, culture flasks, roller bottles, culture slides, and modified Boyden chambers depending on the characteristics of culture cells and culture purpose. Among them, culture dishes and culture flasks or roller bottles are primarily used for cell proliferation purposes, and in some cases, they are also used for study of cell differentiation. Cultured slides are basically used for morphological or some intelligent studies because of the very small size of the slides.
Thus, an artificially made cell culture container may have different surface characteristics from the extracellular matrix originally accommodated in the cells, and cell proliferation and differentiation efficiency may be lowered. Indeed, although various cells have been artificially proliferated and used for clinical treatment, induction of differentiation of various cells including stem cells for treating patients is not easily succeeded.
In recent years, research has been conducted on application of carbon nanotubes to glass and electrodes for stimulation and regeneration of nerve cells in the scientific application field of life science. Two methods of coating are known. First, there is a problem that cytotoxicity due to carbon nanotube aggregation occurs due to direct injection of a carbon nanotube solution into a culture solution. The second method involves placing a carbon nanotube on a glass cover glass, which is subjected to a simple coating or a complex bonding step. In this case, the coated carbon nanotube easily breaks down and the complex bonding reaction is carried out in three or four steps, Time, it is troublesome to coat a large number of glass due to the limit of the glass area when a large amount of cells are cultured.
Accordingly, the present inventors have studied a cell culture container coated with carbon nanotubes capable of improving cell growth without cytotoxicity, and a method of binding functional carbon nanotubes to a cell culture container through simple chemical bonding The present invention has been completed.
It is an object of the present invention to provide a method for manufacturing a cell culture container in which functional carbon nanotubes are coated on the surface quickly and easily.
Another object of the present invention is to provide a cell culture container manufactured by the above-described method.
In order to solve the above problems, the present invention provides a cell culture container comprising a cell culture container having an amine group on the surface of a cell culture container, and a carbon nanotube having a carboxyl group on the surface of the carbon nanotube, Wherein the amine group of the carbon nanotube is amide bonded to the carboxyl group of the carbon nanotube to bind the carbon nanotube to the surface of the cell culture container.
Preferably, the cell culture container further comprises ethylenediamine having an amine group at the terminal, and the amine group of the ethylenediamine and the carboxyl group of the carbon nanotube are amide bonded.
The cell culture container is preferably made of aminated polystyrene.
The carbon nanotubes are preferably multi-walled carbon nanotubes.
The amount of the carbon nanotubes to be added is preferably 20 μg / ml to 30 μg / ml based on 100 μg / ml of the lowest cytotoxicity, and the amount of the ethylenediamine added is 80 mM to 30 mM, 120 mM.
The present invention also provides a method for producing a cell culture container, which comprises binding a carbon nanotube having an amine group to a surface of a carbon nanotube to a cell culture container having an amine group on the surface of the cell culture container.
According to one embodiment of the present invention, the cell culture container to which the functional carbon nanotube is bound has no cytotoxicity, and has an effect of improving cell differentiation and growth.
The cell culture container having the surface coated with the functional carbon nanotubes according to the present invention is not cytotoxic and has an effect of improving the cell differentiation and growth activity.
In addition, proteins or drugs can be bound to carbon nanotubes and used as a new drug delivery support.
BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic view showing a manufacturing process of a cell culture container (a) coated with carbon nanotubes according to an embodiment of the present invention, and FIG.
FIG. 2 is an electron micrograph of the surface of a cell culture container coated with (a) an amine cell culture container and (b) carbon nanotube according to an embodiment of the present invention.
FIG. 3 is a graph showing an electrical resistance measurement result of a cell culture container coated with a carbon nanotube according to an embodiment of the present invention.
FIG. 4 is a graph showing a result of nerve cell toxicity of a carbon nanotube-coated cell culture container according to an embodiment of the present invention.
Hereinafter, the present invention will be described in more detail with reference to the following Production Examples and Examples. However, the following Preparation Examples and Examples are for illustrating the present invention, but the scope of the present invention is not limited thereto.
Example : Fabrication of cell culture vessel coated with functional carbon nanotubes
Functional carbon nanotube lipid nanoparticles (1 mg / 40 ml) containing a carboxyl group in a polystyrene cell culture container (BD PureCoat ® ) having amine groups were added to 0.006 mM ethylene chloride / hydrochloric acid and stirred at pH 3.0 for 3 hours . Functional carbon nanotube lipid nanoparticles having a carboxyl group every 30 minutes were exchanged, and after completion of the reaction, they were washed with distilled water. Then, 0.1 M H 2 N (CH 2 ) 2 NH 2 (ethylenediamine) lipid nanoparticles were added to the obtained amine-carbon nanotube-carboxyl group, and the mixture was added to a 0.012 chloride / hydrochloric acid solution and stirred at pH 5.0 for 2 hours Lt; / RTI > After completion of the reaction, the cells were washed with distilled water and dried to obtain a cell culture container coated with functional carbon nanobube.
Comparative Example : Cell culture container
A commercially available cell culture container (BD PureCoat ™ cell culture plate) was used as a comparative container.
Experimental Example 1: Appearance analysis
The outer shapes of the cell culture containers of the above Examples and Comparative Examples were compared and shown in Fig.
As shown in FIG. 1, the cell culture container coated with the functional carbon nanotubes of the above example exhibited a color different from that of the cell culture container without the carbon nanotube coating of the comparative example. This confirms that the functional carbon nanotubes are bound to the cell culture container.
Experimental Example 2: Electron microscope measurement
The surfaces of the cell culture containers of the above Examples and Comparative Examples were observed through an electron microscope, and the results are shown in Fig.
As shown in FIG. 2, it was confirmed that the carbon nanotubes were uniformly bound to the cell culture container as a single layer.
Experimental Example 3: Measurement of cell resistance
The electrical resistance of the cell culture containers of the above Examples and Comparative Examples was measured, and the results are shown in FIG.
As shown in FIG. 3, the cell culture container of the comparative example had no electrical conductivity at all, and the cell culture container coated with the carbon nanotubes of the example showed electrical conductivity.
Experimental Example 4: Measurement of nerve cell toxicity
The nerve cell toxicity of the cell culture containers of the above Examples and Comparative Examples was measured and the results are shown in FIG.
As shown in FIG. 4, it was confirmed that the growth of neurons was more active in the cell culture container of the carbon nanotube-coated example than the cell culture container of the comparative example in which the carbon nanotubes were not coated. It can be seen that it does not show cytotoxicity.
Claims (7)
Wherein the amine group of the cell culture container and the carboxyl group of the carbon nanotube are amide bonded so that the carbon nanotube is bound to the surface of the cell culture container.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020120138420A KR20140070206A (en) | 2012-11-30 | 2012-11-30 | Cell culture container and the method of preparation thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020120138420A KR20140070206A (en) | 2012-11-30 | 2012-11-30 | Cell culture container and the method of preparation thereof |
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| KR20140070206A true KR20140070206A (en) | 2014-06-10 |
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| KR1020120138420A KR20140070206A (en) | 2012-11-30 | 2012-11-30 | Cell culture container and the method of preparation thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101953472B1 (en) * | 2017-08-22 | 2019-03-04 | 서강대학교산학협력단 | Cell culture chip for modulating direction of neuronal dendrite comprising complex nanostructure and method for manufacturing thereof |
| KR20210069760A (en) * | 2019-12-03 | 2021-06-14 | 단국대학교 천안캠퍼스 산학협력단 | scaffold comprising cabon nanotube interfaced biopolymer nanofiber and preparation method thereof |
-
2012
- 2012-11-30 KR KR1020120138420A patent/KR20140070206A/en not_active Application Discontinuation
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101953472B1 (en) * | 2017-08-22 | 2019-03-04 | 서강대학교산학협력단 | Cell culture chip for modulating direction of neuronal dendrite comprising complex nanostructure and method for manufacturing thereof |
| KR20210069760A (en) * | 2019-12-03 | 2021-06-14 | 단국대학교 천안캠퍼스 산학협력단 | scaffold comprising cabon nanotube interfaced biopolymer nanofiber and preparation method thereof |
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