JPS60244282A - Formed article for tissue culture - Google Patents

Abstract

PURPOSE:To obtain the titled formed article having excellent reusability and the proliferation property of cells, by forming an aromatic polyester copolymer composed of a terephthalic acid component, an isophthalic acid component and a bisphenol component. CONSTITUTION:An aromatic polyester copolymer composed of a terephthalic acid component, an isophthalic acid component and a bisphenol component, wherein the molar ratio of the terephthalic acid component to the isophthalic acid component is 8:2-1:9, is prepared by conventional method. The copolymer may be incorporated with other molding material or a thermal decomposition staibilizer to improve the heat-resistance and oxidation resistance of the polymer. A formed article for tissue culture such as culture flask, petri dish, wide- mouthed bottle, beaker, etc. is formed from the polymer.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a molded article for tissue culture, and more particularly, to a molded article for tissue culture that has excellent cell proliferation, non-contamination with foreign microorganisms, reusability, and operability. Glass culture vessels and culture dishes have been used for tissue culture (hereinafter simply referred to as culture), but recently culture vessels with styrene polymer skeletons and hydrophilic culture surfaces have been developed to improve cell adhesion. is becoming widely used.

Originally, the containers used for tissue culture must be easy and reliable to perform aseptic operations to prevent contamination with contaminants, cells should grow easily and be hard to mutate, and should be transparent and easy to observe. In addition, it would be ideal for culture operations to be easy to handle and economical due to repeated use, but at present, there are no culture vessels that meet all of these requirements, and their appearance is slow. It's long awaited.

In other words, glass can withstand flame sterilization at the mouth to ensure aseptic operation and autoclave sterilization for repeated use.
Although it has advantages such as good proliferation, it is inferior to plastic in formability, so its shape and size are limited. It has drawbacks such as being unable to be transported, easily damaged by impact, and extremely difficult to handle.

On the other hand, polystyrene culture vessels with a hydrophilic culture surface do not have any problems in culture performance or handling, but cannot be flame sterilized due to lack of flame resistance and heat deformation resistance, and are difficult to reuse. When sterilized using pressurized steam using an autoclave, which is the most commonly used method, the product is deformed and becomes unusable, and its reusability is also problematic.

In view of the current situation, the present inventors have developed the growth performance, aseptic operability (flame-resistant sterilization), reusability (autoclave-resistant sterilization), ease of handling, and transparency required for tissue culture vessels. As a result of intensive studies aimed at developing a culture vessel that has all of the properties such as Ta.

Thus, according to the present invention, the composition is composed of a terephthalic acid component, an isophthalic acid component and a bisphenol component, and the molar ratio of the terephthalic acid component to the inphthalic acid component is 8:2 to 1:9.
A molded article for tissue culture is provided by molding an aromatic polyester copolymer.

The aromatic polyester copolymer used in the present invention consists of a terephthalic acid component, an isophthalic acid component, and a hisphenol component. The terephthalic acid component and isophthalic acid component should be understood to include derivatives such as terephthalic acid chloride and isophthalic acid chloride in addition to terephthalic acid and isophthalic acid.

Such a copolymer can be produced according to a conventional method, and specific examples thereof include a solution polymerization method (Japanese Patent Publication No. 37-5599) in which aromatic dicardenoyl chloride and a bisphenol component are reacted in an organic solvent, An interfacial polymerization method in which aromatic dicarboxylic acid chloride dissolved in an organic solvent incompatible with bisphenol component dissolved in an alkaline aqueous solution is mixed and reacted (% Koko No. 40-1959). Examples include a melt polymerization method in which heating is performed in the presence of acetic acid.

In the present invention, the molar ratio of terephthalic acid groups and isophthalic acid groups in the copolymer is 8:2 to 1:9, preferably 7:
It is necessary to maintain a ratio of 3 to 2:8. ◎ If the molar ratio of terephthalic acid groups exceeds 8, the crystallinity of the polymer will increase, resulting in decreased transparency and difficulty in observing the culture state. Also, as a result of the higher melting point, the moldability decreases. On the other hand, if the molar ratio of inteptalic acid groups exceeds 9, only a relatively low molecular weight polymer will be obtained, which will not only lack strength but also have a low melting point, resulting in poor flame sterilization.

The bisphenol component used in the present invention has the following general formula (1
) In the formula, X is 10-, -S-.

-5o2-, -o-o-1 represents an alkylene group that may have a substituent, R1' R2, R5r R4r R
4') R2'.

R3' and R4' are selected from the group consisting of a hydrogen atom, a halodane atom, and a hydrocarbon group.

Examples of such bisphenol components include, for example, 4.4
'-dihydroxy-diphenyl ether, 4.41-
Dihydroxy-2,21-dimethyl-diphenyl ether, 4,4'-dihydroxy-3,3'-dichloro-diphenyl ether, 4,4'-dihydroxy-diphenyl sulfide, 4,4/-dihydroxy-diphenyl sulfone, 4,4/ -dihydroxy-diphenylketone, 4.4/-dihydroxy-diphenyl-methane, 1.
1-(4,4/-dihydroxy-diphenyl)-ethane, 2,2-(4,4/-dihydroxy-diphenyl)-gulonone, 1,1-(4,4/-dihydroxy-diphenyl)-n -butane, (4t4'-dihydroxy-diphenyl)-cyclohexyl-methane, 1.1-
(4,4/-dihydroxy-diphenyl) -2,2
,2-)lichloroethane, etc., but the most commonly produced and representative one is 2.2-(4,4
/-dihydroxy-diphenyl)-pronone, or bisphenol A.

In the present invention, other molding materials may be used together with the aromatic polyester copolymer within a range that does not essentially impede the effects of the present invention.

Specific examples of such materials include 4,4'-dioxydiallylalkane polycarbonate and polyethylene terephthalate. These are usually
It is used in a proportion of 30% by weight or less.

In addition, in order to improve heat resistance, light resistance, and oxidation resistance, thermal decomposition inhibitors, antioxidants, ultraviolet absorbers, etc. can be blended as appropriate. Specific examples include benzotriazole compounds, phosphorus compounds, phenol and benzophenone derivatives.

Examples of molded articles for tissue culture in the present invention include culture flasks, petri dishes, narrow-mouth bottles, wide-mouth bottles, multiwell plates, test tubes, beakers, culture devices in the form of food for mass culture, and carriers for microcarrier complete culture devices. , centrifuge tubes, pipettes, films, sheets, etc., and are particularly useful as molded products that require strong reuse because they can be flame sterilized.

These molded articles can be manufactured by injection molding, blow molding, injection blow molding, extrusion molding, vacuum forming, calendar molding, or any other molding method suitable for forming the intended molded article. In addition, in order to increase the hydrophilicity and cell proliferation of the surface of the molded product, plasma discharge and corona discharge treatments,
The present invention also includes acid treatment and the like.

less than! ! The present invention will be explained in more detail with reference to Examples. Note that parts in the examples are based on weight.

Example 1 11 parts of 2.2-bis(p-hydroxyphenylpropane) (bisphenol A)', 0.2 parts of o-phenylphenol, and 4 parts of calcium hydroxide were dissolved in 250 parts of water, and trimethylpendyl ammonium was added as a catalyst. A solution containing 0.05 part of cum chloride and 7 parts of terephthalic acid dichloride
A polyaromatic polyester copolymer is synthesized by an interfacial polymerization method from a solution of 7 parts of isophthalic acid dichloride (the molar ratio of terephthalic acid groups and isophthalic acid groups is 5:5) in 125 parts of methylene chloride. did. Using a mixed solvent of phenol/tetrachloroethane (6:4) at 25°C (concentration 1
The inherant viscosity (η1nh) measured at #/#) is 0
．． It was 65.

This copolymer is thoroughly dried and extruded using an extruder,
It was cut into pellets. Next, using an injection molding machine and an ultrasonic welding machine whose nozzle temperature was adjusted to 370°C, a tissue culture flask having a mouth was obtained from the pellet. After thoroughly washing these 7 flasks, they were sterilized with high-pressure steam at 120° C. for 20 minutes in a sterilizing autoclave and used for testing. The results are shown in Table 1. The performance evaluation was carried out as follows: (1) Using flame sterilized near-ropan gas, the burner was placed at a distance of 8-8 cm above the tip of the gas burner, which emitted a flame at full throttle. After sterilizing the incubator by rotating it for 10 seconds while holding the mouth part, observe changes in the shape of the mouth part.

(2) Autoclave sterilization: After placing the culture vessel in an autoclave sterilizer and sterilizing it at 120°C for 20 minutes,
Take it out and observe the change in shape.

(3) Chinese hamster normal lung cells (Do
n) to 4 z, z6xxo/d, and this dispersion was aseptically sown into an incubator at a rate of 5.65 x 10'/cm2, and a Teflon sheet was placed on the inside of the sterilized container. Comes with a beautiful phenolic resin cap,
After culturing for 4 days at 37° C. for cell proliferation evaluation, the number of cells in each cell was counted and calculated using a hemocytometer. - (4) Handling A, Stability: Three containers were stacked and their stability was investigated.

31 Strength: The incubator was filled with 20 IIL of water and dropped 5 times from a height of 1.5 m above the concrete surface with the inlet facing up to check for damage.

(5) Suitability for microscopic observation: Cells adhered to the culture surface of the culture vessel were observed using an inverted microscope to evaluate the clarity of cell morphology.

Table 1 *I Nunclon (153732 (Inter
Manufactured by Mad) *2 1611-12OA! Square culture flask (manufactured by Murasaki Vario Glass) Example 2 2.2-Bis(p-hydroxyphenylpropane) (11 parts of bisphenol A was dissolved in an aqueous solution containing 4 parts of calcium hydroxide, and trimethylpene was used as a catalyst. A solution containing 0.05 part of dilammonium coulide, 2.8 parts of terephthalic acid dichloride, and 11.2 parts of isophthalic acid dichloride.
(the molar ratio of terephthalic acid groups and isophthalic acid groups is 2:8)
) was dissolved in 125 parts of methylene chloride to synthesize an aromatic polyester copolymer by an interfacial polymerization method.

This polymer was prepared using the same method as in Example-1.
was 0.61.

A petri dish for tissue culture was molded from the pellets of this copolymer using an injection molding machine with a nozzle temperature adjusted to 370°C, treated in the same manner as in Example 1, and subjected to a test.

- All test methods were performed in the same manner as in Example 1 except for those shown below.

(1) Flame sterilization: The same procedure as in Example 1 was carried out except that the lid of the petri dish was held horizontally with tweezers, and changes in shape were observed.

(2) Cell proliferation: Cells were seeded in the same manner as in Example 1, covered with a lid, and cultured in a carbon dioxide atmosphere at 37° C. for 5 ts. The same procedure as #1 was carried out.

(3) Handling properties: strength The lid was kept horizontal in a closed state and dropped repeatedly from a height of 1.5 m above a concrete surface 5 times to check for damage.

Table 2 *1 Polystyrene 3002 (60x15m) (
BoCton, Dlckinmon aIlld Co.
Example 3 9.8 parts of terephthalic acid dichloride, 4.2 parts of in7thalic acid dichloride (mole of terephthalic acid group and isophthalic acid group) An aromatic polyester copolymer was synthesized in the same manner as in Example 2, except that the ratio was 7:3). The obtained copolymer (4) had ηinh of 0.65. This copolymer (4) was formed into pellets in the same manner as in Example 1 and subjected to molding.

In addition, 75 parts of this copolymer (A) and 4,4'-dioxyallylalkane polycarbonate (4) Gunlite L
-1225', manufactured by Yondai Kasei Co., Ltd., 25 parts were mixed,
It was extruded using an extruder and cut to obtain pellets of mixed resin (B), which were similarly subjected to molding.

The above two types of pellets were used separately to form tissue culture mettles using an injection molding machine and an ultrasonic welding machine, and were subjected to the same treatment as in Example 1 and subjected to testing. The test method is shown below. Everything except the point plate was carried out in the same manner as in Example 1. (1) Cell proliferation: Cells were seeded and the sealed container was kept at 37°C.
, as in Example 1, except for rotating culture (at 15 RPH).

Table 3

Claims (1)

[Claims] 1. It is formed by molding an aromatic polyester copolymer consisting of a terephthalic acid component, an intermolecular component, and a bisphenol component, and the molar ratio of the terephthalic acid component and isophthalic acid component is 8:2 to 1:9. Molded products for tissue culture.