WO2011096659A2

WO2011096659A2 - Microorganism or cell culture container

Info

Publication number: WO2011096659A2
Application number: PCT/KR2011/000440
Authority: WO
Inventors: 전민용
Original assignee: Jeon Min-Yong
Priority date: 2010-02-05
Filing date: 2011-01-21
Publication date: 2011-08-11
Also published as: WO2011096659A3; KR20110091078A

Abstract

The present invention relates to a container for culturing microorganisms or cells, wherein a part or the entire culture container are formed with an elastic membrane and/or a filter membrane comprising rubber, silicone or the like. The culture container comprises: a body of a container having a throughhole on one surface; and the elastic membrane for stopping the throughhole of the body of a container, comprising an elastomer, wherein the elastic membrane can be a dual layer, a sterilization layer can be included, and a filter capable of ventilating air can only be added.

Description

Microbe or Cell Culture Vessel

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a microorganism or a cell culture vessel, and to a culture vessel that blocks invasion of bacteria or various bacteria and reduces contamination by using an elastic membrane in a culture propagation apparatus such as a microorganism or a living tissue.

Culture growth of biological materials is the most fundamental work of bio industry such as identification and amplification of DNA as well as material analysis in microbiology genetic engineering and medicine.

In general, aseptic culture of all animal and plant cells and tissues is injected into a sterile culture vessel, the sterile cells and tissues are healed therein, and then filled with a stopper to prevent external contamination. It is a so-called subculture that is cultured for a period of time, and when the culture is grown after a certain period of time and the space in the culture container is physically insufficient, or when the amount or moisture of the culture medium is depleted, the culture is changed to fresh culture. It is done periodically.

Modern medicine is trying to obtain various therapeutic effects using stem cells. Stem cells include embryonic stem cells from early blastocysts (Blstocyst) and adult stem cells from adult or placenta after the developmental process. There are three main methods of obtaining these adult stem cells: first through bone marrow, second through umbilical cord blood, and then peripheral blood. easily obtained through mesenchymal blood. In addition, techniques for obtaining adult stem cells from adipocytes or other tissues of the body have been researched and developed.

However, since the stem cells are collected in such a small number, they make a large amount of stem cells through culture and inject them into tissues that require cell regeneration such as skin, cardiovascular, and organ tissues. The collected stem cells are inoculated and cultured in culture vessels of various shapes and types by traditional culture methods.

The conventional method of culturing strains or cells is to sterilize the inoculation needle with an alcohol lamp on a work bench that is blocked from outside air called cleanbench in a laboratory, and then remove the lid or stopper of the culture vessel containing the strain or biological tissue to be inoculated. After opening, use a needle to remove some of the strains, cells, etc., the chalena triangle flask containing the culture solution, or the culture vessel with the lid or stopper of the culture vessel or tissue culture well or plate open. After plating on the culture medium, the inoculation process is performed by sealing or covering the lid or stopper of the culture vessel.

The culture process of such a conventional microbial operation is briefly described with reference to FIG.

Inoculation and incubation process of the microbial strain of the conventional method, as shown in Figure 1, sterilize the tweezers or inoculation needle in the flame of the alcohol lamp in the open space, open the lid (2) of the cell (1) containing the cell culture medium The strain in (17) was buried in the inoculation needle (3), and then the lid (2) of the chalet (1) to be cultured was opened to spread the strain on the inoculation needle (3) onto the medium (17). Then, the lid 2 is covered with a tape sealing process. Subsequent cultures are incubated for a certain period of time in an incubator capable of controlling the environment, that is, the temperature or the atmosphere of the stem cells to be grown, and the medium is replaced or subcultured as necessary. The process is repeated.

However, in the culture propagation process, bacteria attached to the surface of the air or work tool in the working space during the process of opening and closing the lid of the culture vessel, inoculating stem cells, or injecting the culture solution, Cells or spores of various bacteria can be infiltrated into the culture vessel. Therefore, in such a culture propagation process, a lot of costs are consumed to control the contamination of the culture according to the skill of the operator or the contamination of the work space.

Therefore, in order to prevent this, microbiological laboratories are equipped with a microbiological aseptic workbench, which occupies a large space called a clean bench, and even though work is performed in such a space, work failure occurs due to contamination of the culture proliferation caused by the contamination of the microorganisms described above. have.

Patent Publication No. 2001-0029122 discloses an animal and plant cell culture vessel for preventing microbial contamination, which is equipped with a special micro filter that does not pass external pollutants and can easily exchange gas to prevent microbial contamination in the culture process. Suggesting.

Patent Publication No. 1020040051602 discloses a container and a cap for a container provided with a filter medium for aeration of a container and a filter medium for aeration, in order to prevent entry of germs or mites into a medium in a sterilized culture container. Ventilation filter media for

Iii) and a cultivation vessel with such media and a cap for the cultivation vessel is shown. This cultivation bottle is provided with a bottle main body and the cap attached to an opening part. The cap has a cap body and a filter medium for ventilation. The cap body is provided with a hole. As a filter medium for aeration, a porous membrane made of polytetrafluoroethylene and having micropores having a diameter of 0.1 to 50 µm or less is provided.

In order to prevent contamination, clean benches, which are microbiological aseptics, are equipped with a device that blocks the inflow of external air and prevents and sterilizes microorganisms in the air or the fallout of its spores. In this small medical institution, there is a difficulty that cannot be provided. Even if a medical institution, a microbiological laboratory, or a bio-industrial equipped with such equipment is present, contamination of the tissue culture during microbial or stem cell cultivation depending on the skill of the operator or the degree of microbial contamination in the clean bench. This is inevitable, which wastes time and increases work cost.

Since the cleanbench forms an open space in a large volume device, an expensive cleanbench with complicated facilities is being developed to increase the efficiency of the pollution prevention function. However, even these expensive and complicated clean benches cannot be completely enclosed spaces where microorganisms, which are the source of contaminants, can not enter during continuous operation. Many times. Thus, there is a need for a highly skilled workforce and a high probability of failure, resulting in lower productivity and increased costs.

Literature Information of the Prior Art

[Document 2] KR 10-2001-0029122 (Korea Research Institute of Bioscience and Biotechnology) 2001. 4. 6

[Reference 3] KR 10-0629588 (Daikin High School Co., Ltd.) September 27, 2006

[US 4] US 2010/0015694 A1 (CARLO ACOSTA) 2010. 1. 21

In the present invention, it is possible to improve the convenience of operation in the apparatus for inoculating and cultivating animal or plant tissues, strains and stem cells, to prevent contamination of microorganisms that may occur in the working process, and to cultivate economically and efficiently cells. To provide a culture vessel.

The present invention is different from the conventional method of opening and closing a container containing a culture medium or a strain and a flora and fauna tissue, and contaminating cells and microorganisms due to the inflow of microorganisms present in the air by injecting or withdrawing the contents without opening and closing the culture vessel. To prevent it.

The present invention is to provide a culture vessel capable of performing aseptic microbial and cell manipulation even in the absence of a clean bench.

In the present invention, in order to reduce the biological effort generated by the highly skilled research personnel to reduce the biological contamination generated during microbial and stem cell work and to reduce the loss of working time caused by microbial contamination, Even if you want to be able to easily carry out culture propagation of microorganisms and stem cells without contamination.

The present invention is to provide a culture vessel for storing the culture medium in a powder state in order to increase the shelf life of the culture medium container.

The present invention is intended to be able to quantitatively inject a strain or a solution in a closed state through the elastic membrane of the culture vessel with the tip of the pointed micropipette in order to inject the correct amount into the culture vessel.

The present invention exports the gas generated from the cells or strains in culture, the required gas is introduced into the culture vessel to block the infiltration of microorganisms or their spores that cause contamination of the culture, the external contaminating microorganisms or their spores It is prevented from being penetrated into the culture vessel by being buried on the surface of the operating device such as the needle inserted into the microorganism or the inoculation needle micropipette tip, and even though the microorganism penetrated through the primary elastic membrane is blocked by the secondary elastic membrane. Since the contact with the sterile layer is limited is to provide a culture vessel that is naturally inhibited and killed.

In the present invention, a rubber, silicone, or the like is used in the body or stopper of a culture vessel such as a culture medium, an enzyme, a reagent, a culture medium or a container containing, collecting or culturing microorganisms or biological tissues, that is, a tissue culture well or chalet, an Erlenmeyer flask, a culture bottle, or a plate. Forming an elastic membrane or elastic stopper formed by using the same, and using the operating mechanism, such as inoculation needle, needle or scraper microneedle tip passing through the elastic membrane or elastic stopper is collected and collected by the culture solution, bacteria, biological tissues, etc. Sealing operation of the culture medium or animal or plant tissues, strains, stem cells, etc. transplanted and propagated to be made.

The present invention is a culture vessel for culturing a strain or cells, comprising a container body having a through hole on one surface, and an elastic membrane made of an elastic body blocking the through hole of the container body.

In the present invention, the elastic membrane is fixedly attached around the through-hole or coupled in a detachable stopper form, or the elastic membrane is a laminated elastic membrane laminated with two or more elastic membranes, a sterilization layer between the elastic membrane of the laminated elastic membrane Is formed.

According to the present invention, a through hole is formed in a part of the container body, and a filter is attached to block the through hole.

The elastic membrane of the present invention has a filter that blocks a part of the through hole and blocks the remaining unblocked part of the through hole.

The culture vessel of the present invention has a three-dimensional structure according to the type of culture strain or cells that have three or more small chalena or cell culture wells formed inside the container body, and three-dimensional uneven treatment on the inner bottom of the container body. One containing a three-dimensional cell culture artificial material inside the container body, coated with a material that aids the attachment of cells to the inner surface of the container body, or containing a solid or liquid medium inside the container body will be.

According to the present invention, electrical or magnetic energy is applied to the inside of the container body, or the elastic membrane serves as a filter and the elastic membrane at the same time, or a partition wall is formed on the bottom of the container body.

According to the present invention, the tip of the micropipette for injecting or withdrawing a certain amount of material through the elastic membrane has a sharply shaped tip, or the needle portion of the micropipette tip is formed of a metal material and attached to the micropipette or the pipette. The part is formed of synthetic plastic.

The present invention is to keep the culture solution in a powder or dried state in the container body, sterile distilled water, physiological saline, or other solvent through the elastic membrane immediately before inoculating microorganisms or cells to be used.

Stem cell culture vessel to prevent the contamination of microorganisms according to the present invention has the following effects.

First, without opening and closing the container containing the culture solution or strains and animal and plant tissues and withdraw the contents to prevent the contamination of stem cells and microorganisms due to the inflow of microorganisms present in the air.

Second, by inoculating the contents of the culture medium or strain and animal and plant tissues without opening and closing the container to prevent contamination of stem cells and microbial culture proliferation due to the inflow of microorganisms in the air.

Third, the culture and proliferation of strains, animal and plant tissues and stem cells in an enclosed space prevents contamination that can occur in open spaces, occupies a large space and eliminates sterility even in the absence of a clean bench, which increases the cost burden. Microbial and stem cell manipulation operations can be performed.

Fourth, to reduce the biological pollution caused by microbial and stem cell work, the highly skilled research personnel should pay close attention to the mental effort and reduce the loss of working time caused by microbial contamination. There is an effect of preventing the rise of the part, even non-skilled people can easily perform culture and growth operations such as microorganisms and stem cells without contamination.

* Fifth, by injecting sterile distilled water, physiological saline or other solvents through the elastic membrane immediately before inoculating microorganisms or animals or cells in the culture vessels stored in powder form, the shelf life of the culture vessels can be extended.

Sixth, the tip of the tip of the micropipette can penetrate the elastic membrane of the culture vessel and quantitatively inject the strain or solution in a closed state.

Seventh, the gas generated from the cells or strains in culture is discharged, and the required gas flows into the culture vessel, and at the same time a filter is formed to block the infiltration of microorganisms or spores that cause contamination of the culture, thereby preventing contamination in microbial culture. Block it.

Eighth, by forming a plurality of layers of elastic membrane, it is fundamentally to prevent the external contaminating microorganisms or their spores are buried on the surface of the operating mechanism, such as the needle or inoculation needle micropipette tip inserted when the microorganisms are manipulated.

Ninth, the presence of microorganisms penetrating through the primary elastic membrane due to the coating or formation of the germicidal layer between the elastic layers of the plurality of layers, even if the microorganisms infiltrate the secondary elastic membrane and limited infiltration, the contact with the germicidal layer, the activity is naturally Suppressed and killed.

1 is an exemplary view showing a process of a conventional microbial operation

2 to 11 illustrate one embodiment of the present invention.

※ Explanation of code about main part of drawing ※

1: chalet, 2: lid, 3: inoculation needle

4: elastic membrane, 5: through hole, 6: adhesive layer,

7: stopper, 8: needle, 9: micropipette tip,

10: groove, 11: ring, 12: jaw,

13: second elastic film, 14: sterile layer, 15: filter,

16: container body 17: medium 19: micropipette

2 and 3 is a view showing a first embodiment of the culture vessel having an antifouling elastic membrane of the present invention. In the drawings, components or parts having the same function are denoted by the same reference numerals for convenience of understanding.

The present invention forms a through hole in one side of the lid 2 or the body 16 of the chalet 1, which is a culture vessel, and is closed with an elastic membrane 4. It is formed in at least one through-hole 5 formed in the body 16 or the lid 2 of a synthetic plastic or glass material. The medium 17 is installed in the body 16.

The elastic membrane 4 is usually made of elastic material such as rubber, silicone, or the like, and is bonded around the through hole 5 formed on either side of the body 16 or the lid 2 of the culture vessel. The edge portion of the elastic membrane is fused with the container by heat, ultrasonic waves, or high frequency fusion to form an adhesive layer 6. The material of the elastic membrane is not limited to rubber or silicone, and the elastic membrane 4 may be made of a thin film as a material having elastic force. Even if a puncture occurs by the needle 8 or the like, the puncture is closed again to close the culture vessel. You can make it using recoverable materials. The elastic membrane 4 can also be made of synthetic plastics that act as filters. In addition, the elastic membrane 4 may be formed to have a composite function in which rubber or silicon is coated on a portion thereof. In addition, the elastic membrane 4 may be formed in a fine fiber shape so as to perform the role of the filter 15 at the same time.

FIG. 4 shows another embodiment of the present invention, showing that the elastic membrane 4 is coupled to the through hole 5 of the lid 2 in the form of a stopper 7. The elastic membrane 4 may be coupled to the through hole of the container body 16 or the lid 2 and fitted into a hole formed in a portion of the elastic membrane 4 fixedly attached to the through hole 5. May be combined. The stopper 7 may be detachably coupled to the lid 2 or a part of the container body 16 and a part of the stopper 7 in a fused form, or the stopper 7 may be attached to a part of the elastic membrane 4. A part of) may be formed by fusion. In this embodiment, the same function as in the first embodiment except that the plug can be opened and closed.

5 is a view showing another embodiment of the present invention, in which the plurality of chalets 1 are disposed in the body 16 of the culture vessel, and the lid covering the body corresponds to the position of each chalet. Stopper 4 is formed in the position.

6 is a view showing another embodiment of the present invention, in which the entire opening portion is sealed with the elastic membrane 4 instead of the lid of the culture vessel body 16. Of course, before sealing, it is provided with the necessary medium 17 and various components necessary for cultivation and then sealed with the elastic membrane 4. In this example, in order to ensure the adhesion of the elastic membrane 4 to the open edge of the main body, it is necessary to make the jaw 12 to put the adhesive area with the elastic membrane 4 on the edge and to form the adhesive layer 6 thereon. good.

In the embodiment of FIGS. 5 and 6, the elastic membrane 4 has a needle 1 through which the needle 8, the inoculation needle 3, or the tip 9 of the micropipette 19 is sharply formed like a needle. Inoculate strains, animals and plants and stem cells into the culture medium (17) or supplement the culture solution, etc., and remove the holes so that the perforated holes are elastically closed. Function.

On the other hand, in general, the end cross section of the conventional micropipette tip 9 is rounded or cut at an angle of 90 degrees, in the present invention, the micropipette tip 9 penetrates the elastic membrane 4 to the chalet 1 The tip is sharply inclined so as to penetrate the inside, like a syringe needle, so that the elastic membrane 4 can be easily penetrated. The material of the micropipette tip 9 may be made of synthetic plastic or metal material, or the end portion penetrating and penetrating the elastic membrane 4 is formed of metal and inserted into the micropipette 19 or the pipette. The losing part may be formed of synthetic plastics such as PE, PP, and PET. In addition, the groove 10 is formed so that the rubber or silicone ring 11 is fitted to the side edge of the micropipette or the pipette, and the sealing ring 11 is fitted to engage the micropipette tip 9 so as to strongly bind the pipette or the micropipette. So that it does not fall easily during microbial manipulation.

Penetrating the elastic membrane 4 is not limited to the needle (8) inoculation needle (3) micropipette tip (9) described above, in the culture vessel such as chalets (1), Erlenmeyer flasks, culture bottles, tissue culture wells, etc. Instruments that are deemed necessary for a variety of manipulation operations, such as micro-tweezers, micro-scissors, knives, scrapers, etc., are operated without opening the cap (7) or lid (2) of the culture vessel outside the culture vessel. Any apparatus that can be used for the work can be used. That is, through the elastic membrane 4 of the culture vessel with a small hole, the instruments used for the operation work are put into the container, and inoculated and injected with the cells of the required strain or animal or plant or the necessary culture solution, cell differentiation inducing solution, enzyme reagent, or the like. Alternatively, the microorganisms and stem cell manipulation operations can be performed in conventional culture vessels or culture vessels by performing an operation for separating and transplanting cultured cells or strains and replenishing the necessary culture solution in a closed culture vessel. Compared to contaminated through the open and close operation of the open and closed process, the contamination is significantly reduced, eliminating the cost and waste of time, and also reducing the need for a clean bench that takes up a large amount of space. It is very convenient to use in hospitals, schools, laboratories, etc. All.

Figure 7 is a view showing another embodiment of the present invention, the culture vessel body 16 is used to have a form such as a bottle, the lid 2 is a structure to be sealed using a bottle cap. In addition, the through-holes are formed on one side of the body, and then the elastic membrane 4 is attached, and the through-holes are formed on one side of the body for the distribution of gas necessary for cultivation, and the filter 15 is attached. The filter may be a microfilter that allows gas and gas generated at the time of cultivation to be used for the strain or cell to be cultured. The medium 17 is installed on the lower side to which the elastic membrane and the filter are not attached. This embodiment shows that the elastic membrane 4 can be formed in the body 16 of the culture vessel as well as the lid 2 according to the shape and use of the culture vessel.

8 is a view showing another embodiment of the present invention, the culture vessel has a body 16 having a plurality of culture cell chalets and a lid 2 formed with an elastic membrane over a wide range. This lid is provided with the filter 15 superimposed on the elastic membrane 4.

The culture vessel may be a round or square dish as well as a triangular flask or bottle type or a type of cell culture plate in which a plurality of compartment cells or wells are formed. In addition, when it grows better in three-dimensional casting depending on the culture strain or cell type, it is good to increase the surface area by widening the surface area by applying the three-dimensional design irregularities on the bottom surface, or the three-dimensional culture condition. have. In addition, the inner surface of the culture vessel may be coated with a material that helps the adhesion of the cells, it may be in the form of a culture vessel to induce three-dimensional cell proliferation by putting a three-dimensional artificial material inside.

The elastic membrane 4 may be formed on the lid 2 of the double culture vessel in which a plurality of chalets 1 are disposed, and the medium 17 of the liquid or powder may be pre-injected. In addition, such a culture vessel may form a ring of silicone or rubber material between the culture vessel and the lid 2 in order to increase the sealing effect between the body 16 and the lid 2, and the screw or coupling formed on the body and the lid 2 The jaws can be attached to each other. Such a culture vessel may be in the form of a plurality of the above-described cell culture wells, or alternatively, may be in the form of a culture vessel containing a plurality of containers. In addition, the culture may be well performed by applying electric energy or magnetic energy from the outside of the culture vessel to the inside of the vessel.

In addition, although the elastic membrane 4 is not shown, a culture medium container for storing the culture medium which affects the cell growth of microorganisms or animals or plants, differentiation culture medium which induces differentiation of various cells, enzymes and reagents, etc. It may be applied to the stopper portion of the reagent vessel and formed. Although a stopper formed of a material such as plastic or glass is contained in a culture container containing a conventional culture solution, there is a possibility of causing microbial contamination during the opening and closing of the stopper to obtain its contents, and a reagent containing a liquid enzyme or reagent. The same holds true for containers. In addition to the above liquid contents, the lyophilized medium 17 component in powder form may also be applied. In this case, by injecting sterile distilled water or physiological saline or other solvents through the elastic membrane 4 into a culture container stored in a powder state. The shelf life of the culture vessel can be extended. Therefore, the culture vessel or the reagent vessel sealed with the stopper 7 formed with the elastic membrane 4 of the present invention can be used to remove the contents using the needle 8 or the tip 9 of the micropipette without opening the stopper 7. It is possible to reduce the microbial contamination through such a closed operation process because it can be applied or injected into the culture vessel, or on the contrary to inject the required solvent components in the culture vessel.

9 and 10 show another embodiment of the present invention. In this example, the elastic membrane 4 is formed on the film-shaped filter 15 attached to the adhesive layer 6 around the through hole 5 formed in the lid 2. A filter 15 is attached for the release of the gas generated during the at least one microbial metabolism of the culture vessel or the body of the culture vessel and the growth of the flora and fauna and the introduction of the required gas. At this time, the filter 15 is formed of a material having a permeability enough to block the influx of microorganisms or spores of the pollutant and to distribute the gas generated during the growth and cultivation of cultured cells or strains. On the other hand, a portion of the filter 15 may be formed by coating the elastic membrane 4.

Although not shown, a membrane made of a material having gas permeability and elastic force at the same time may be formed as the elastic membrane 4 or the filter 15 to form part or the whole of the culture vessel.

Figure 11 shows another embodiment of the present invention. In this example, the lid 2 portion of the culture vessel containing the medium 17 is fused or adhered to the body 16 of the container by the elastic membrane 4 as a whole. At this time, the elastic membrane 4 is attached to the jaw 12 formed on the body 16, the elastic membrane through which the operating mechanism, such as the needle (8), inoculation needle (3), micropipette tip (9) ( 4) The second elastic film 13 is added to the site to form a double. It is better to form a film coated with a disinfectant in the double layer of elastic membrane to disinfect the needle entering and exiting the layer.

In this way, the elastic membrane 4 through which the operating mechanisms such as the needle 8, the inoculation needle 3, and the micropipette tip 9 penetrates is doubled, and microorganisms that may be buried on the surface of the operating mechanism, Spores, etc. are primarily blocked by the external second elastic film 13, or the remaining spores or microorganisms passing therethrough are once again blocked by the elastic membrane 4 formed below. In addition, even if microorganisms infiltrated through the first elastic layer 13 by the sterilization layer 14 provided between the double

elastic membranes

4 and 13 exist, they are blocked by the elastic membrane 4 and contaminated with limited contamination. Since the microorganisms come into contact with the bactericidal layer, their activity is naturally inhibited and killed. This structure is to inject a certain amount of medium (17) into the culture vessel automatically in the factory for the efficiency of mass production, and the second elastic membrane ( When the edge portion of the elastic membrane 4 having 13) is fused or bonded, production efficiency is improved. The culture vessel thus formed has the effect of blocking the contaminating microorganisms close to perfection, thereby reducing the space of the clean bench which occupies a large volume. It can be done.

The microorganism or cell culture apparatus for preventing the microbial contamination of the present invention is not limited to the above-described embodiment, anyone of ordinary skill in the art without departing from the gist of the present invention and various modifications can be carried out It will be referred to the technical scope of this invention to the extent possible.

The present invention relates to a microorganism or a cell culture vessel, and provides a culture vessel that blocks invasion of bacteria or various bacteria and reduces contamination by using an elastic membrane in a culture propagation device such as a microorganism or a living tissue. It can be used in the medical industry, various laboratories, or the manufacture of medical devices. Culture growth of biological materials is the most fundamental work of bio industry such as identification and amplification of DNA as well as material analysis in microbiology genetic engineering and medicine. Aseptic culture of cells or tissues is designed to prevent external sources of contamination. Since the work is important, the present invention can be usefully used for this purpose. In particular, modern medicine is doing a variety of treatments using stem cells, which can be useful for culturing stem cells.

Claims

As a culture vessel for culturing strains or cells,

Container body having a through hole on one side,

Culture vessel comprising an elastic membrane made of an elastic body to block the through-holes of the container body
The culture vessel according to claim 1, wherein the elastic membrane is fixedly attached around the through hole or coupled in a detachable stopper shape.
The culture vessel of claim 1, wherein the elastic membrane is a laminated elastic membrane laminated with two or more elastic membranes.
The culture vessel of claim 3, wherein a sterilization layer is formed between the elastic membranes of the laminated elastic membrane.
The culture vessel according to claim 1, wherein a through hole is formed in a part of the container body, and a filter is attached to block the through hole.
The culture vessel of claim 1, wherein the elastic membrane has a filter that blocks a portion of the through hole and blocks the remaining portion of the through hole.
The culture vessel according to claim 1, wherein at least two small chalena or cell culture wells are formed in the container body.
The culture vessel according to claim 1, wherein the culture vessel has a three-dimensional structure according to the type of culture strain or cells subjected to three-dimensional unevenness on the inner bottom of the container body.
According to claim 1, Culture vessel characterized in that the artificial material for cell culture three-dimensional in the interior of the container body.
The culture vessel according to claim 1, wherein a coating material is coated on the inner surface of the container body to assist cell adhesion.
The culture vessel according to claim 1, wherein a solid or liquid medium is contained in the container body.
The culture vessel of claim 1, wherein electrical or magnetic energy is applied to the interior of the vessel body.
The culture vessel of claim 1, wherein the elastic membrane serves as a filter and an elastic membrane at the same time.
The culture vessel according to claim 1, wherein a partition is formed at the bottom of the container body.
The culture vessel of claim 1, wherein the tip of the micropipette for injecting or withdrawing a certain amount of material through the elastic membrane has a sharply shaped tip.
16. The culture vessel of claim 15, wherein the needle portion of the micropipette tip is formed of metal and the portion attached to the micropipette or pipette is formed of synthetic plastic.
The method according to any one of claims 1 to 16,

The culture vessel is characterized in that the culture medium is stored in the container body in a powder or dried state, and sterile distilled water, physiological saline, or other solvents are introduced through an elastic membrane immediately before inoculating microorganisms or cells.
A culture vessel characterized in that part or all of the culture vessel for culturing strains or cells is made of a material that acts as a filter and an elastic membrane