JPS6143984A - Formed article for tissue culture - Google Patents

Abstract

PURPOSE:To provide the titled article having high adhesivity to cells without using the hydrophilic treatment of the surface, and enabling the stable growth and prolideration of cells for a long period, by molding a polyarylate. CONSTITUTION:The objective article can be manufactured by the injection molding of a blent polymer obtained by blending e.g. 30pts. of a polycarbonate with 70pts. of a polyarylate derived from (A) an aromatic dicarboxylic acid preferably composed of terephthalic acid and isophthalic acid at a molar ratio of 7/3-3/7 and (B) a dihydric phenol of formula I [R1-R4 are H, halogen or (halogenated) hydrocarbon group] or formula II [R1'-R4' are same as R1-R4; X is single bond, O, S, SO, CO, (halogen-substituted) alkylene or (halogen-substituted) alkylidene] [e.g. 2,2-bis(4-hydroxyphenyl)propane, etc.].

Description

[Detailed description of the invention] The present invention relates to a molded article for tissue culture.

In recent years, transparent synthetic resin containers and films have come into widespread use in place of glass as molded articles for tissue culture.

These synthetic resins are generally polystyrene, polymethyl methacrylate, polyvinyl chloride, or modified products obtained by copolymerization thereof. However, molded articles for tissue culture using these synthetic resins lack hydrophilicity, and if left as is, the adhesion of cells to such molded articles is poor and desirable tissue culture cannot be achieved.

Therefore, conventionally, various methods have been used to treat the surface of molded products.
Attempts have been made to increase its hydrophilicity. For example, hydrophilic resin coating, ozone treatment.

Corona discharge treatment, treatment with oxidizing agents or mineral acids, plasma treatment, β-ray irradiation, treatment with alkaline treatment agents, sandblasting treatment2, etc. are already known. It is difficult to obtain stable data on cell viability and proliferation due to a large decrease in hydrophilicity over time and repeated use, and cells that have once attached to the surface of a molded product often peel off from the surface during growth. They have common drawbacks such as:

Furthermore, depending on the treatment method, toxicity to cells on the surface of the molded article cannot be completely eliminated, and there is also a drawback that the growth state of the cells varies depending on the type of cells.

Furthermore, conventionally used molded articles for tissue culture generally have a low heat deformation temperature, so when used for tissue culture they are usually sterilized with ethylene oxide, but this requires time and effort.
Moreover, it is a cause of cost increase.

As a result of intensive research aimed at solving the problems of tissue culture molded products using conventional synthetic resins as described above, the present inventors surprisingly succeeded in molding polyarylate into tissue culture molded products. If a surface-treated product is used, cell adhesion is good even after hydrophilic treatment, and therefore there is no instability seen with conventional products with surface treatment, and stable data on cell growth and proliferation can be obtained over a long period of time. The present invention was based on the fact that the present invention can be obtained.

That is, the present invention is a molded article for tissue culture formed by molding a polyarylate.

The polyarylate used in the present invention is obtained from an aromatic dicarboxylic acid or a functional derivative thereof and a dihydric phenol or a functional derivative thereof.

The aromatic dicarboxylic acid used in the preparation of the polyarylate may be any aromatic dicarboxylic acid as long as it reacts with dihydric phenol to give a satisfactory polymer, and may be used alone or in combination of two or more. A preferred aromatic dicarboxylic acid is terephthalic acid.

Isophthalic acid is mentioned, and mixtures thereof are particularly preferred in terms of melt processability and overall performance.

In the case of such a mixture, the mixing ratio is not limited, but is terephthalic acid/isophthalic acid -9/1 to 1/9.
(molar ratio) is preferably 1. Particularly from the viewpoint of melt processability and performance balance, 7/3 to 3/7 (molar ratio), and more preferably 1/1 (molar ratio) is divalent used for the preparation of polyarylates. As phenol, the following general formula (1), (I
t) or (III).

In the above general formula, RI + R! ＋R! *R
4R'+, R'x, R'3 + R'4 are selected from the group consisting of hydrogen atom, halogen atom, hydrocarbon group, and halogenated hydrocarbon group, and X is o, s, so, co, alkylene group Alternatively, an alkylidene group (if necessary, the alkylene group or alkylidene group may be substituted with one or more halogen atoms).

) represents. Specific examples of preferred dihydric phenols include 2
．． 2-bis(4-hydroxyphenyl)propane, 2.
2-bis(4-hydroxy-3,5-dibromophenyl)propane, 2.2-bis(4-hydroxy-3,5-dichlorophenyl)propane, 4.4'-dihydroxydiphenylsulfone, 4.4' -dihydroxydiphenyl ether.

4.4'-dihydroxydiphenyl sulfide 54.
4′-dihydroxydiphenylketone, 4.4′-
Dihydroxydiphenylmethane, 2,2'-bis(
4-hydroxy3,5-dimethylphenyl)propane,
1.1-bis(4-hydroxyphenyl)ethane, 1.
1-bis(4-hydroxyphenyl)cyclohexane,
4.4'-dihydroxydiphenyl.

Examples include benzoquinone. These may be used alone or in combination of two or more. Although these dihydric phenols are para-substituted, other isomers may be used, and ethylene glycol, propylene glycol, etc. may be used in combination with these dihydric phenols. The most typical dihydric phenols are
2.2-bis(4-hydroxyphenyl)propane,
It is called JLAt bisphenol 8, and is most preferable from a comprehensive viewpoint.

Therefore, the most preferred polyarylate in the present invention is a mixture of terephthalic acid and isophthalic acid or functional derivatives thereof (provided that the molar ratio of terephthalic acid groups to isophthalic acid groups is 9:1 to 1:9, particularly 7 :3 to 3 nia) and a dihydric phenol represented by the general formula (III), particularly bisphenol A or a functional derivative thereof.

In the present invention, not only polyarylate alone but also a resin composition of polyarylate can be used. The other component constituting the resin composition may be one that has good compatibility with the polyarylate, provides a transparent resin composition, and does not adversely affect cell growth and proliferation. Anything can be used preferably, but
Particularly preferred are polycarbonate and polyethylene terephthalate. Such a resin has the effect of improving the moldability of polyarylate.

When using a resin composition of polyarylate and polycarbonate, if the weight fraction of polyarylate is less than 50%, cell viability and proliferation will decrease, so the weight fraction of polyarylate is preferably 50% or more. When using a resin composition of polyarylate and polyethylene terephthalate, the weight fraction of polyarylate is preferably 70% or more for the same reason.

In the present invention, molded products include containers such as petri dishes, multi-well plates, narrow-mouth bottles, wide-bottle bottles, flasks, testers, and beakers, as well as films and sheets, and these include injection molding, blow molding, extrusion molding, and calendering. It can be manufactured by an appropriate molding method such as molding.

The tissue culture molded article of the present invention has a hydrophilic surface that is good for all cell lines that are generally used as subjects for tissue culture tests, but representative cell lines that can be suitably cultured include, for example, He1a ( human cervical cancer), L (
mouse normal subcutaneous connective tissue).

[-38 (human normal fetal lung), KB (hydrobasal layer).

Chang Liver (human normal liver 11i), P
L (human normal sheep II), Don (Chinese hamster normal lung).

Examples include BHK-21 (Syrian hamster normal kidney).

The molded article for tissue culture of the present invention uses polyarylate, and unlike conventional surface-treated articles, its hydrophilicity does not decrease over time because it is not treated, and therefore has good cell growth and proliferation properties. In addition to good performance, stable data can be obtained over a long period of time.

Furthermore, since it does not undergo any thermal deformation at temperatures of about 130° C., it has the characteristic that it can be sterilized by dry heat or autoclave like a glass container when performing tissue culture.

The present invention will be explained in more detail below by giving examples.

Examples 1 to 3 ■Boriaarylate (manufactured by Unitika ■, U-polymer U-10
0), ■Boriaarylate (same as above) and polycarbonate,
(Kijin Kasei Panlight L-1250) and 70
/30 (fI weight ratio) and a resin composition consisting of 80/20 (weight ratio) of polyarylate (same as above) and polyethylene terephthalate (manufactured by Unitika, MA2103) were injection molded and directly molded. 75s+m
Got a chalet. Place this chalet in a heated oven for 1
It was sterilized under ventilation at a temperature of 20° C. for 1 hour and then subjected to tissue culture. Note that the chalet was not thermally deformed at all during this sterilization process.

Eegle MEM medium 10+wj! Bovine
5eru+w 2mj! and 7.5% NaHCO32
Use 5rag of a nutrient solution prepared by adding m It as a medium,
To this, He1aS-3 cells (manufactured by Dainippon Pharmaceutical tII) IX
After administering a solution containing IO'' cells (approximately 1-1), culturing was carried out under heat at 37°C in a stream of 5% carbon dioxide using chalets each containing 31!L of each of the above ■, ■, and ■. 48 hours after the start of culture, 0.25% trypsin solution was added to the chalet, and the cells that had grown on the wall of the chalet were detached from the wall to form a cell suspension.The cell concentration in this cell suspension was determined by Burker-Turk. When measured using a type hemocytometer, the cell concentration in the medium was approximately 1.4 x 10' per ml, approximately 1.5 x 10' and approximately 1.5 per ml.
In the case of X10B pieces, approximately 0.9X10B 1 Approximately 1. OX
10" and approximately 1.5X10" pieces, approximately 0.9 in the case of ■
X101i + approximately 1.0X10'' and approximately 1.1X10', stable data were obtained for each, and it was confirmed that the cells multiplied approximately 7.5 to 5 times in the width of the chalet.

Next, after washing the chalet that was used once above.

The cells were sterilized and repeated culture tests were conducted in the same manner as above. As a result, the case of ■ is 1.4X10".

Approximately 1.5X10'' and approximately 1.6X10B pieces;
lX10'' pieces, ■ case is approximately 1.lXl0'l approximately 1.
Data of 1X10" and about 1.2X to' tlJ were obtained, and it was confirmed that stable values could be changed even after repeated use.

Patent applicant: Unitika Co., Ltd. VCT -AAF,-

Claims (1)

[Claims] (1) A molded article for tissue culture made of polyarylate.