JPH01165367A - Cell cultivation container and method for treating inner surface thereof and device therefor - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION [Object of the Invention] (Industrial Application Field) The present invention relates to a cell culture container and a method and apparatus for treating its inner surface.

(Prior Art) In recent years, so-called culturing in which microorganisms, tissue cells, etc. are divided and multiplied has been widely practiced, and in particular, for culturing adherent cells, various containers specially treated to improve cell adhesion are used.

In particular, in order to increase the adhesion area of cultured cells and improve culture efficiency, recently, as shown in FIG. is used, the medium and cells are placed in the container 1, placed horizontally, and the body 1b of the container is rotated.
Some devices are designed to allow cultured cells to adhere to the entire inner peripheral surface of the cell.

On the other hand, since the inner surface of an untreated plastic bottle-shaped container has a low negative charge density, it is difficult for cells to adhere to the inner surface, and therefore there is a problem in that good implantation is not achieved.

One of the surface treatment means for increasing the negative charge density on the surface of such plastics is a plasma treatment technique using glow discharge. This plastic treatment is generally carried out either at high temperatures or at room temperature under a high magnetic field, but if the target container is made of plastic with a low point, plasma processing at high temperatures may cause the container to melt. This results in deformation and cannot be applied, so plasma treatment using glow discharge at room temperature and under a high magnetic field is an effective method.

This surface treatment technology using glow discharge at room temperature and high magnetic field is
Electrodes 3, 4 are placed in the vacuum chamber 2 as schematically shown in FIG.
are placed opposite each other, and a container 1 to be treated is placed between these electrodes 3 and 4.
', while a working gas 5 such as oxygen or an inert gas is blown in from the side and a discharge is caused between the electrodes 3 and 4 to activate the activated working gas to increase the negative charge density on the inner surface. be able to.

(Problems to be Solved by the Invention) However, according to the above-mentioned conventional plasma treatment using glow discharge, if the container 1' is dish-shaped, such as a Petri dish, as shown in FIG. However, in the case of a bottle-shaped container having a narrow mouth portion 1a as shown in FIG. Even if surface treatment is carried out on the outer circumferential surface of the body and the central part of the inner surface of the bottom of the bottle-shaped container, the surface treatment of the thin line area, that is, the essential inner surface of the body of the bottle-shaped container, is almost impossible. There was a problem in that the entire inner surface could not be treated uniformly, making the container unsatisfactory as a cell culture container.

To improve this, US Patent No. 4,238,568
There is a technique shown in the specification of No. In order to uniformly apply surface treatment to the entire inner surface of the bottle-shaped container, this product has a three-part joint structure consisting of a cylindrical body, a bottom plate, and an upper lid including a mouth, and a plasma treatment stage. In this method, the above three parts are disassembled and processed individually, and after processing, they are assembled to form a bottle-shaped container.According to this method, the bottle-shaped container can be produced at low cost and with high efficiency by integral molding such as blow molding. However, injection molding and welding processes are required, which makes the bottle-shaped container itself expensive, and an extra assembly process is required.

In view of this, the present invention provides a culture container that has an appropriate negative charge density on the inner surface even if it is a bottle-shaped container having a narrow mouth that is integrally molded in advance, and a culture container that has a uniform negative charge on the inner surface. The object of this invention is to provide a surface treatment method and apparatus capable of carrying out density-increasing treatment.

[Structure of the invention]

(Means for Solving the Problems) In order to solve the problems of the prior art described above, the present invention has a narrow opening integrally formed by blow molding or the like, and a cap can be tightly fitted into the opening. The first invention is a cell culture container characterized in that the inner surface of the bottle-shaped container is treated to increase negative charge density by glow discharge. A high-frequency electrode is inserted from the bottle-shaped container, a working gas such as oxygen or inert gas is supplied, and the high-frequency electrode is energized to generate a glow discharge inside the bottle-shaped container, thereby increasing the negative charge density on the inner surface of the bottle-shaped container. The second invention is a method for treating the inner surface of a cell culture container, characterized in that the inner surface of the cell culture container is
It is equipped with a high-frequency electrode wound into a spiral coil shape with an outer diameter that can be inserted from the mouth of a bottle-shaped container using a hollow tube, and a working gas supply system such as oxygen or inert gas is provided to the hollow tube constituting the electrode. At the same time, a gas ejection port is opened in the part constituting the spiral coil-shaped high frequency electrode, and a working gas is ejected into the bottle-shaped container through the electrode, and plasma is generated by energizing the high frequency electrode. The third invention provides an apparatus for treating the inner surface of a cell culture container, characterized in that the negative charge density on the inner surface of the container is increased.

(Function) Since the entire inner surface of the bottle-shaped container is made to have an appropriate negative charge density by plasma treatment using glow discharge,
By rotating the bottle-shaped container, the cells and the like inside the bottle-shaped container reliably adhere to the inner surface of the bottle-shaped container, thereby increasing the culture efficiency.

In addition, in the method and apparatus according to the present invention, the high-frequency electrode is inserted into the interior of the bottle-shaped container through the mouth and discharge is caused from inside the container, so that uniform plasma treatment can be performed over the entire inner surface of the bottle-shaped container. No dead zone occurs regardless of the inner shape of the container.

(Example) Hereinafter, the present invention will be explained using an example shown in FIGS. 2 and 3, using the same reference numerals for parts common to those in FIG. 4.

In the method for treating the inner surface of a cell culture container according to the present invention, a bottle-shaped container 1 to be treated is placed in a vacuum chamber 2 with its mouth 1a facing downward, as shown in a schematic cross section in FIG. A high-frequency electrode 6 is inserted through the opening 1a, and a working gas such as oxygen or an inert gas is supplied to energize the high-frequency electrode 6, thereby generating a high magnetic field 7 around the electrode 6. A process is performed in which a glow discharge is caused to generate plasma, and the negative charge density is increased over the entire inner surface of the bottle-shaped container 1.

Therefore, even if the container is a bottle-shaped one having a narrow mouth 1a, the entire inner surface can be treated evenly, and the bottle-shaped container 1 can be placed horizontally and cultured while being rotated. When used as a roller bottle, cells can be attached to the entire inner surface of the container.

In order to perform the above-mentioned glow discharge even better and to simplify the configuration of the working gas supply system, the high-frequency electrode 6 is shown in use in FIG. 2, and partially enlarged in FIG. There is. That is, the hollow tube material 9 is wound into a spiral coil shape so as to have an outer diameter that can be inserted through the mouth portion 1a of the bottle-shaped container 1, and the gas outlet 10 is opened at the tip of the spiral coil electrode portion 6a. A gas supply pipe 11 is connected to a portion of the hollow tube material 9 located outside the bottle-shaped container 1 . In the figure, reference numeral 12 indicates a gas supply source, and reference numeral 13 indicates a high frequency power source connected to the high frequency electrode 6 via the matching device 8. Therefore, since the high-frequency electrode 6 has a spiral coil shape, the actual length of the electrode is long and glow discharge can be performed well, and the working gas can be supplied through this electrode, so the gas supply system The structure is simplified, and since it is only necessary to insert the high-frequency electrode 6 into the bottle-shaped container 1, the workability of the inner surface treatment work is greatly improved. Note that the gas outlet 11 is not limited to the tip of the spiral coil electrode 6, and may be opened in the middle.

〔Effect of the invention〕

As explained above, in the cell culture container according to the present invention, the negative charge density on the inner surface of the container is increased by plasma treatment using glow discharge, so that the adhesion of cells to the inner surface of the container is increased during cell culture. , cells can be cultured with high efficiency.

In addition, in the surface treatment method according to the present invention, a high-frequency electrode is inserted into the container from the mouth of the container, and a working gas is supplied into the container to perform plasma treatment by glow discharge. Even if it is a bottle-shaped container, the negative charge density can be increased evenly over the entire inner surface of the container.
Therefore, the inner surface can be treated regardless of the shape of the container, and since the inner surface can be treated after the container itself has been formed in advance, there are no restrictions on the manufacturing of the container, which increases the cost of the container. It is possible to obtain a culture vessel with high efficiency without inviting. Furthermore, the inner surface treatment device according to the present invention has a structure in which a high-frequency electrode is inserted into the interior from the mouth of a bottle-shaped container and is wound in a spiral coil shape with a vibrator, and a working gas such as oxygen or an inert gas is passed through the vibrator. Since the gas is supplied, glow discharge can be performed well, and at the same time, the structure of the gas supply system is simplified, and the structure of the entire apparatus is simplified.

[Brief explanation of the drawing]

FIG. 1 is a cross-sectional view showing an example of a cell culture container according to the present invention, FIG. 2 is a cross-sectional view showing an example of an apparatus for implementing the inner surface treatment method according to the present invention, and FIG. 3 is a cross-sectional view showing an example of the same high-frequency electrode. FIG. 4 is a sectional view showing a conventional glow discharge device, and FIG. 5 is a sectional view of a container showing problems with this device. DESCRIPTION OF SYMBOLS 1... Bottle-shaped container, 1a... Mouth part, 16... Body part, 2... Vacuum chamber, 6... High frequency electrode, 9...
...Hollow pipe material 1.10...Gas outlet, 11...Gas supply pipe. Applicant's agent Mr. Sato

Claims (1)

[Claims] 1. The inner surface of a plastic bottle-shaped container has a narrow mouth integrally formed by blow molding or the like, and a cap can be tightly fitted into the mouth. A cell culture container characterized by being subjected to charge density increasing treatment. 2. The cell culture container according to claim 1, wherein the container has a rollable body. 3. In the vacuum chamber, insert a high-frequency electrode from the mouth of the bottle-shaped container, supply a working gas such as oxygen or inert gas, and energize the high-frequency electrode to cause glow discharge inside the bottle-shaped container. 1. A method for treating the inner surface of a cell culture container, comprising subjecting the inner surface of the bottle-shaped container to negative charge density increasing treatment. 4. Equipped with a vacuum chamber and a high-frequency electrode wound into a spiral coil shape with an outer diameter that can be inserted from the mouth of a bottle-shaped container using a hollow tube, and oxygen and inert gas are applied to the hollow tube constituting the electrode. A working gas supply system such as the above is connected, and a gas outlet is opened in the part constituting the spiral coil-shaped high-frequency electrode, and the working gas is jetted into the bottle-shaped container through the electrode to supply the high-frequency electrode to the high-frequency electrode. 1. A device for treating the inner surface of a cell culture container, characterized in that the inner surface of the container is subjected to negative charge density increasing treatment by generating plasma through energization.