JPH08149973A - Culture container - Google Patents

Abstract

PURPOSE: To provide an inexpensive culture container, comprising a film excellent in light transmittance, humectant properties, durability, high gas permeability, heat adhesion, etc., hardly having contaminating properties, capable of providing an environment suitable for culturing a cell, a tissue, an organ or the whole individual of an animal, a plant or a microorganisms, etc., and attaining excellent culture efficiency, especially culture of a plant requiring a large quantity of gaseous carbon dioxide for photosynthetic reaction, above all, excellent culture efficiency in the culturing in a photoautotrophic system without containing any sugar in a culture medium. CONSTITUTION: This culture container is obtained by using a film, comprising a polymer containing 4-methyl-1-pentene or a butadiene polymer and having gas permeabilities of 5,000-80,000 (cm<3> /m<2> .D.atm) oxygen gas permeability (PO2 ) and 15,000-250,000 (cm<3> /m<2> .D.atm) gaseous carbon dioxide permeability (PCO2 ) when formed into 30μm thickness in at least a part of the container.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a culture container, and more specifically, it comprises a film excellent in high gas permeability, durability, moisture retention, thermal adhesiveness, etc. The present invention relates to a culture container that can be inexpensively provided with a suitable culture environment for cells, tissues, organs or the whole of animals and plants or microorganisms, and can achieve excellent culture efficiency.

[0002]

2. Description of the Related Art A culture vessel provides various culture properties to cells, tissues, organs or whole individuals of animals and plants or microorganisms, and therefore has various properties such as light permeability, breathability, moisture retention, durability and chemical resistance. It is required to satisfy the requirements such as sex.

By the way, conventionally, culture vessels made of glass or various resins have been used. However, since these culture vessels have poor air permeability, oxygen and oxygen necessary for respiration of the culture and photosynthesis are used. There is a drawback in that carbon dioxide becomes starved and the growth of the culture becomes poor. In order to avoid such problems, loosen the screw cap and use it, or use a cotton plug, paper stopper, aluminum foil, etc. for gas exchange, but there are still cases where sufficient ventilation cannot be secured. There are many drawbacks, and there is a high risk of invasion of various bacteria.

Therefore, there is a demand for a culture vessel which retains various properties such as light permeability, air permeability, moisture retention, durability, and chemical resistance, while having sufficient air permeability without invasion of various bacteria. . In order to meet such a demand, for example, Japanese Patent Laid-Open No.
Japanese Patent Laid-Open No. 3-198972 proposes a culture container using a fluororesin. However, higher gas permeability is required for some culture targets or conditions, and in addition, the fluororesin has a melting temperature of about 30.
Since it is as high as 0 ° C., there is a problem that the heat-bonding operation at the time of producing the culture container is not easy. Further, there is a problem that the fluororesin is relatively expensive.

Further, Japanese Patent Laid-Open No. 3-277268 discloses a cell culture bag using a film made of a polymer alloy of low density polyethylene and linear low density polyethylene. However, the gas permeability of low density polyethylene is not sufficient.

[0006]

SUMMARY OF THE INVENTION The present invention has been made in view of such circumstances, and an object thereof is to have excellent light transmittance, moisture retention, durability, high gas permeability, thermal adhesiveness and the like. It is composed of a film, and is less contaminated, and at a low cost, provides an environment suitable for culturing cells such as animals and plants or microorganisms, tissues, organs or whole individuals, and can achieve excellent culturing efficiency, In particular, to provide a culture vessel capable of achieving excellent culture efficiency for culturing a plant that requires a large amount of carbon dioxide gas for a photosynthetic reaction, and particularly for culturing in a photoautotrophic system that does not contain sugar in the medium. is there.

[0007]

That is, the gist of the present invention consists of a polymer containing 4-methyl-1-pentene or a butadiene polymer and has an oxygen gas permeability (PO ₂ ) when molded to a thickness of 30 μm. ) Is 5,000-80,000
(Cm ³ / m ² · D · atm), carbon dioxide permeability (PC
O ₂ ) is 15,000 to 250,000 (cm ³ / m ²
A culture container characterized by comprising a film having a gas permeability of D.atm) at least in a part of the container.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in detail below.
The film used in the culture vessel of the present invention is made of a polymer containing 4-methyl-1-pentene or a butadiene polymer. In the present invention, as a polymer containing 4-methyl-1-pentene, a homopolymer of 4-methyl-1-pentene, a copolymer with 4-methyl-1-pentene, 4-
A graft polymer of a homopolymer of methyl-1-pentene,
It includes a mixture of a polymer of 4-methyl-1-pentene and another polymer.

Other monomers in the copolymer with 4-methyl-1-pentene include α-olefins having 2 to 24 carbon atoms such as ethylene, propylene, 1-butene,
1-hexene, 1-octene, 1-decene, 1-tetradecene, 1-octadecene, 1-hexadecene, 1-dodecene, 1-tetradodecene and the like can be mentioned. Preferably, 1-hexadecene, 1-octadecene and the like are used. One or more of these α-olefins are copolymerized with 4-methyl-1-pentene. The amount of α-olefin in the copolymer is 0.1 to 20% by weight, preferably 1 to 15% by weight.

The copolymer with 4-methyl-1-pentene may be a graft polymer having a 4-methyl-1-pentene homopolymer as the main chain. As the monomer to be grafted, a modified monomer such as (meth) acrylic acid, maleic acid, fumaric acid, tetrahydrophthalic acid,
Unsaturated carboxylic acids such as itaconic acid, citraconic acid, crotonic acid, isocrotonic acid, and nadic acid, or derivatives thereof, such as acid halides, acid amides, acid imides, acid anhydrides, esters and the like can be mentioned. And, as specific examples of the above-mentioned derivatives, malelen chloride, maleimide, maleic anhydride, citraconic anhydride, dimethyl maleate,
Examples thereof include glycidyl maleate and (meth) acrylic acid alkyl ester.

These may be contained alone or in combination of two or more. Of these, unsaturated dicarboxylic acids or acid anhydrides thereof are preferable, and maleic acid, nadic acid or acid anhydrides thereof are particularly preferable. The graft amount is usually 0.0
It is 5 to 20% by weight, preferably 0.1 to 10% by weight. A film using a graft polymer having a homopolymer of 4-methyl-1-pentene as a backbone is excellent in adhesiveness and sealing property.

As the other polymer in the mixture of the 4-methyl-1-pentene polymer and the other polymer, the above-mentioned α-olefin polymer is preferable, and the mixing amount thereof is
0.1 to 20% by weight, preferably 0.5 to 15% by weight
Is.

In the present invention, the butadiene polymer may be a homopolymer or a copolymer thereof, for example, a copolymer with styrene, and a copolymer having 50% by weight or more of styrene is particularly preferable. Further, 1,2-polybutadiene is preferably used as the homopolymer of butadiene.

In the present invention, if necessary, a plasticizer, a heat stabilizer, an antioxidant, a lubricant, an antiblocking agent, an ultraviolet absorber, a coloring agent, or the like may be added to the above-mentioned polymer which is a raw material of the film.
A modifier, an antibacterial agent, an antifungal agent, an antifogging agent, an ethylene removing agent and the like may be added. The method for producing the film is not particularly limited, and specifically, for example, an ordinary inflation method, T-die extrusion method or the like is adopted. The thickness of the film is usually 20 to 50 μm, preferably 25 to 3
It is set to 5 μm. The gas permeability can also be adjusted by adjusting the film thickness, and an optimum gas environment can be provided according to the gas requirements of the culture.

The film of the present invention has a thickness of 30 μm.
Has an oxygen gas permeability (PO ₂ ) of 5,000 when molded into
0-80,000, preferably 10,000-70,
And the carbon dioxide gas permeability (PCO ₂ ) is 15,
000-250,000, preferably 40,000-
250,000 [All units are cm ³ / m ²
・ D.atm (23 ° C)]. In particular, for a culture requiring a large amount of carbon dioxide gas such as a culture in a photoautotrophic system, the carbon dioxide gas permeability is 100,000 to 250,000 [cm ³ /
m ² · D · atm (23 ° C)] is preferably used.

Since the film of the present invention can be completely adhered easily with a commonly used heat sealer, the number of steps for producing the film container is smaller than that of the fluororesin film. In addition, the production costs of the raw material resin and the film are also low, and it is judged that the practicability is obviously high in terms of cost. Moreover, since these films can be easily processed by means such as welding, adhesive bonding, and molding, a culture container having an arbitrary shape can be easily manufactured. The shape of the culture container of the present invention is not particularly limited, but examples thereof include a shape such as a square shape, a cylindrical shape, and a bag shape.

The culture vessel may be composed of one component, and two or more separable components, for example, the culture vessel body and a frame attached to the outside of the body, or the culture vessel body, the frame and the opening. It may be composed of a part sealing device or the like.

An example of the embodiment of the culture container of the present invention will be described with reference to FIG. However, the present invention is not limited to the mode of the container in FIG. 1 or the manufacturing method. In FIG. 1, 1 is a film, 2 is a frame, 3 and 3'are sealing parts, and 4 is a support.

The end of the film 1 is heat-sealed to form a bag having a sealing portion 3, and a frame 2 is installed therein to form a container. Next, the support 4 impregnated with the medium is put in from the upper part of the container, the culture is inoculated, and then the upper part is folded and heat-sealed to form the sealing part 3 '. The sealing portion 3'can completely block invasion of various bacteria.

Alternatively, the support 4 may be set on the frame 2 in advance and inserted into the bag. After the film 1 is mounted on the inside of the frame 2, the bag is prepared by heat-sealing appropriate portions of the film. It may be in a shape. Furthermore, after inserting the frame 2 into the tubular film 1, the lower portion of the film may be heat-sealed to form a bag. The material of the frame 2 is not particularly limited, but resin or the like is usually used.

Since the culture vessel of the present invention has extremely high oxygen gas permeability and carbon dioxide gas permeability, and also has excellent moisture retention and light permeability, it is possible to sufficiently exchange gas during the culture, thereby facilitating animal and plant life. Alternatively, it is suitable for a culture container for cells, tissues, organs or whole individuals of microorganisms. Furthermore, it is preferably used for culturing plants in a photoautotrophic system in which a carbon source such as sugar is removed from the medium under a high carbon dioxide atmosphere. As the culture device, a normal culture device equipped with a carbon dioxide gas supply pipe, a lighting device and the like is used. The culture vessel of the present invention is generally used in a culture device.

A general medium is used in the culture container of the present invention. Examples thereof include White medium, Murashige & Skoog medium, Heller medium, Vacin & Went medium, and modified mediums thereof. If necessary, a plant hormone, an antibiotic, etc. may be added to the medium.

In culturing a photoautotrophic plant in a high carbon dioxide atmosphere, a modified medium obtained by removing a carbon source such as sugar from the above medium is preferably used. In the case of such photoautotrophic culture, propagation of various bacteria is extremely small, and further, surprisingly, carbon dioxide can be fixed and used as a nutrient source. Therefore, the method of cultivating a photoautotrophic plant under a high carbon dioxide atmosphere can ensure good growth equivalent to that of sugar culture and is practically excellent.

In the case of culturing a photoautotrophic plant, the concentration of carbon dioxide gas in the culture device is suitably 500 to 5,000 ppm, preferably 1,000 to 4,000 ppm. The amount of light varies depending on the plant, so it cannot be generally said, but it is usually 1,000 to 6,000, preferably 1,
About 500 to 5,500 lux is suitable. The light source is not particularly limited, and a normal fluorescent lamp or the like is used.

The target plant to which the culture container of the present invention can be applied is not particularly limited, but preferably, orchid, lily, carnation, gypsophila, gerbera, cymbidium, anthurium, spatiferum, and other petals, strawberry, tomato, cucumber, etc. Fruits and vegetables.

[0026]

EXAMPLES The present invention will be described in more detail with reference to examples below, but the present invention is not limited to the following examples unless it exceeds the gist. The oxygen gas permeability and the carbon dioxide gas permeability in the examples were measured at 23 ° C. using a pressurized gas permeability measuring method (using Gasperm-100 type manufactured by JASCO Corporation). In addition, the leaf color is measured by a simple chlorophyll meter (SPAD-502 type manufactured by Minolta).
Was done using.

Examples 1 to 2 1,2-polybutadiene (Example 1, "JSR R820" manufactured by Japan Synthetic Rubber Co., Ltd.) and poly-4-methyl-
1-Pentene (Example 2, "TP manufactured by Mitsui Petrochemical Industry Co., Ltd."
X MX001 ”) was used to produce a film having a thickness of 30 μ by an inflation method. The oxygen gas permeability (PO ₂ ) and carbon dioxide gas permeability (PCO ₂ ) of these films were measured at 23 ° C. The results are shown in Table 1.
All showed high gas permeability. Using these films and a 7.5 × 7.5 × 10.5 cm resin frame, the culture vessels of Examples 1 and 2 having the shape shown in FIG. 1 were produced.

Comparative Examples 1-2 In place of the 1,2-polybutadiene (Japan Synthetic Rubber Co., Ltd. "JSRR820") film in Example 1,
Fluorine resin (Comparative example 1, "PFA" manufactured by Daikin Industries, Ltd.)
30 obtained from the 25 μm film and low density polyethylene (Comparative Example 2, “F131” manufactured by Mitsubishi Kasei)
Oxygen gas permeation rate (P) of each of these films was measured in the same manner as in Example 1 except that a film of μm was used.
O ₂ ) and carbon dioxide permeability (PCO ₂ ) were measured at 23 ° C. The results are shown in Table 1. Furthermore, the culture containers of Comparative Examples 1 and 2 were produced using these films.

[0029]

[Table 1] 1,2PB: 1,2-polybutadiene P4M1P: poly-4-methyl-1-pentene PFA: fluororesin F131: low density polyethylene

Culturing Test 1 Using the culture vessels of Examples 1 and 2 and Comparative Examples 1 and 2, Cymbidium Melody was used.
Fair'Marylyn Monroe 'Expansion Leaf 3
Aseptic culture of 25 to 30 mm of seedlings was performed. The culture method is as follows.

That is, the medium support was made of rock wool multi-block (Gordania AS Denmark).
("AO 18/30" manufactured by the same company) was used, and 100 ml of Vacin & Went medium containing 2% of sucrose per 4 x 4 block was added.
After placing the seedlings of Cymbidium, the seedlings were aseptically sealed, placed in a culture device, and cultured at 25 ° C.

A fluorescent light for growing plants (Plant Torx manufactured by Toshiba Lighting & Technology Co., Ltd.) was used for illumination, and the 16-hour photoperiod was 1,
Cultured at 700 lux. The results of culturing for 98 days are shown in Table 2.
Shown in In addition, as Comparative Example 3, the same culture was performed using a conventionally used culture container (PCB) made of polycarbonate.

The seedlings cultivated in the culture vessels of Examples 1 and 2 showed superior growth to the seedlings cultivated in the culture vessel of Comparative Example 1. The growth of the roots was promoted remarkably and the leaf color was also excellent.
In particular, the culture container of Example 2 was excellent.

[0034]

[Table 2]

Cultivation Test 2 In the same manner as Cultivation Test 1, anthurium seedlings (Anth
urium andreaanum'Elizabeth 'developed leaves 3 leaves, seedling height 20 mm) was sterilized and cultivated in Examples 1-2.
And it compared with the culture container of Comparative Examples 1-3. The medium is Murashige & Skoog.
g) Of the composition of the medium, the amount of calcium chloride and magnesium sulfate was adjusted to half, and 2% sucrose was added to the medium, and the cells were cultured for 69 days. The results are shown in Table 3.

[0036]

[Table 3]

Cultivation Test 3 In Cultivation Test 1, Vacin and Went modified medium containing no sucrose and seedlings of Spathiphyllum were used. Carbon dioxide was supplied into the culture device to raise the concentration to 3,000 ppm. The culture was performed under the same conditions as the culture test 1 except that the culture was performed for 60 days. The results are shown in Table 4. Under this condition, the plant cannot absorb the carbon source from the medium, and assimilates carbon dioxide gas in the culture vessel to perform photoautotrophic growth. Therefore, the gas permeability in the culture vessel, particularly the carbon dioxide gas permeability, greatly affects the growth of plants.

The seedlings cultivated in the culture vessels of Examples 1 and 2 were in very good growth condition, no contamination was observed, and the culture vessels of Comparative Examples 1 and 2 were used for both the above-ground part and the root part. The growth of. In particular, the growth of roots when the culture vessels of Examples 1 and 2 were used was about 2.5 times that when the culture vessel of Comparative Example 1 was used, and the culture vessel of the present invention is photoautotrophic. It has been found that it has an extremely excellent effect on.

[0039]

[Table 4]

[0040]

According to the present invention described above, it is possible to provide an environment suitable for culturing cells, tissues, organs or whole individuals such as animals and plants or microorganisms, and achieve excellent culturing efficiency, In particular, there is provided a culture container capable of achieving excellent culture efficiency for culturing in a photoautotrophic system containing no sugar in the medium.

[Brief description of drawings]

FIG. 1 is a perspective view showing an example of a culture container.

[Explanation of symbols]

 1: Film 2: Frame 3: Sealing part 3 ': Sealing part 4: Support

Claims (1)

[Claims] 1. A polymer containing 4-methyl-1-pentene or a butadiene polymer, which has an oxygen gas permeability (PO ₂ ) of 5,000 to 8 when molded to a thickness of 30 μm.
50,000 (cm ³ / m ² · D · atm), carbon dioxide permeability (PCO ₂ ) 15,000 to 250,000 (c
A culture container comprising a film having a gas permeability of m ³ / m ² · D · atm) as at least a part of the container.