JPS5953030B2 - Moldings for tissue culture - Google Patents

Description

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

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

Synthetic resin molded articles generally lack hydrophilicity, and if left as is, cells will have poor adhesion to such molded articles, making it impossible to perform desired tissue culture.

Therefore, various methods have conventionally been used to treat the surface of molded articles to improve their hydrophilicity.

For example, hydrophilic resin coating, ozone treatment, corona discharge treatment, treatment with oxidizing agents or mineral acids, etc. are already known.

However, with these conventional surface treatments, hydrophilicity deteriorates significantly over time, making it difficult to obtain stable conditions for cell growth and proliferation.Also, cells that have once attached to the surface of the molded product can be molded during growth. They have common drawbacks such as peeling off from surfaces.

Furthermore, depending on the treatment method, toxicity to the 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 tissue culture molded products generally have a low heat distortion temperature, so when used in tissue culture tests, they are usually sterilized with ethylene oxide, but this is time consuming and costly. It can also cause up.

The present invention was made in order to solve the above problems, and the hydrophilicity of the surface does not deteriorate over time. Therefore, preferred tissue culture is possible, and cell growth and proliferation are stable over a long period of time. The object of the present invention is to provide a tissue culture synthetic resin molded product that can be sterilized by dry heat or autoclave because of its high heat distortion temperature.

The tissue culture molded article of the present invention is made of a styrene-maleic anhydride copolymer, and is characterized in that its surface is treated with an alkaline treatment agent.

The styrene-maleic anhydride copolymer used in the present invention has a maleic anhydride content of about 1 to 40 mol%,
The copolymer can be used alone or as a mixture with other synthetic crumbled fats such as polystyrene, or synthetic rubbers such as styrene-butadiene rubber, acrylonitrile-styrene-butadiene rubber, polybutadiene, and polyisoprene.

If too much of these synthetic resins or synthetic rubbers are mixed,
The compatibility with the styrene-maleic anhydride copolymer deteriorates, making it difficult to obtain a good molded product, and generally reduces the heat resistance of the molded product.
0% by weight or less, and in the case of synthetic rubber, 10% by weight or less of the mixture.

In addition, alkaline processing agents include alkali metal hydroxides,
In addition to solutions such as aqueous solutions of alkaline earth metal hydroxides and alkali metal carbonates, and ammonia water, gaseous substances include ammonia gas and mixtures of ammonia gas and water vapor.

The treatment conditions are not particularly limited as long as the alkali treatment imparts sufficient hydrophilicity to the surface of the molded product and preferably does not cause devitrification of the molded product, but usually sodium hydroxide, potassium hydroxide, 10% by weight in an aqueous solution of calcium hydroxide, sodium carbonate, etc.
The molded product is immersed for 1 to 120 minutes at a temperature of ~60°C, washed with water, and dried.

In the present invention, molded products include containers such as petri dishes, multiwell plates, narrow-mouth bottles, wide-mouth bottles, flasks, test tubes, and beakers, as well as films and sheets, and these include injection molding, blow molding, extrusion molding, It can be manufactured by an appropriate molding method such as calendar 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 He1a ( human cervical cancer), L (mouse normal subcutaneous connective tissue), WI-38 (human normal fetal lung)
, KB (Hydroacidalgia), Chang Liver (
Examples include normal human liver), FL (normal human amnion), DOn (normal Chinese hamster lung), and BHK-21 (normal Syrian hamster kidney).

Unlike conventional surface treatments, the tissue culture molded article of the present invention is alkali-treated, and its hydrophilicity does not deteriorate over time. Stable data can be obtained over a period of time, and since it does not undergo any thermal deformation at temperatures of about 120°C, it can be sterilized by dry heat or autoclave like glass containers when culturing tissue.

Example: Injection molded styrene-maleic anhydride copolymer (Dylarc #332 manufactured by Block Plastics Co., Ltd.) Diameter 75 m
After immersing the Petri dish of No. m in a 1% by weight aqueous sodium hydroxide solution at room temperature for 30 minutes, it was washed with water and dried.

The petri dish thus treated with alkali was sterilized in a heated oven at a temperature of 120° C. under ventilation for 1 hour, and then subjected to tissue culture.

In addition, in this sterilization process, the Petri dish was not thermally deformed at all.

Add Bovine se to 10ml of Eegle MEM medium.
He1a S
After administering a solution (approximately 1 ml) containing 1×10 5 -3 cells (manufactured by Dainippon Pharmaceutical ■), the cells were cultured in a 5% carbon dioxide gas stream while keeping the temperature at 37°C.

48 hours after the start of culture, a 0.25% trypsin solution was added to the Petri dish, and the cells grown on the Petri dish wall were detached from the wall to form a cell suspension.

The cell concentration in this cell suspension is determined by Burker-Tur.
When measured using a type K hemocytometer, it was confirmed that the concentration of cells in the medium was about 1 x 105 cells per ml, and that the cells had proliferated about 5 times in the petri dish.

Claims (1)

[Scope of Claims] 1. A molded article for tissue culture comprising a styrene-maleic anhydride copolymer, the surface of which is treated with an alkaline treatment agent. 2. The tissue culture molded article according to claim 1, wherein the alkaline treatment agent is an aqueous sodium hydroxide solution.