JPH0833480A - Plant cell cultivation apparatus - Google Patents

Abstract

PURPOSE:To provide a plant cell cultivation vessel enabling the injection of chemicals without causing environmental pollution. CONSTITUTION:This plant cell cultivation apparatus is composed of a culture vessel 1 and a lid 2 and has a ventilation hole 4. The apparatus is further provided with an injection port sealed during cultivation and enabling the attachment of a chemical injection syringe through a septum seal as necessary and a dust-collection bag attaching port 6 sealed during cultivation and enabling the attachment of a dust collection bag connected to the space in the vessel through a septum seal as necessary. A chemical can be injected into the vessel with simple operation without causing the scattering of the chemical.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a plant cell culture apparatus suitable for cell culture in a narrow room or in a microgravity environment and capable of drug injection without fear of environmental pollution.

[0002]

2. Description of the Related Art A petri dish-type culture device as shown in FIGS. 6A and 6B is used as a device for conducting a plant cell culture test. This device consists of a culture container 1 and a container top lid 2.
In the culture container 1, a medium such as an agar medium is provided on a substrate such as the honeycomb sheet 10, and plant cells are planted in the medium to conduct a culture experiment. During the culture, the outside air is taken in through the ventilation hole 4 provided in the container upper lid 2 through the air-permeable filter (air-permeable membrane 18) such as the Teflon filter, whereby the cells breathe and are cultured. At the end of the culture for a predetermined time, cells are taken out and various tests are performed.For example, when performing a culture test in a shuttle in space, the culture place and the test place are separated and the test is performed immediately after the culture is completed. In many cases, it is not possible to do so or it takes some time before the test due to some reason. In such a case, if left as it is, further culturing progresses, and accurate data cannot be obtained. Therefore, a method of injecting a drug such as glutaraldehyde into the container to stop the culturing progress is adopted. This work is called fixed work.

[0003]

When injecting a drug into such a culture device in a laboratory on the ground, the lid of the petri dish is removed and the drug is injected. However, in this case, since the device is not sealed during the injection work, a toxic drug may leak out into the room, and in a microgravity space such as outer space, the contents are scattered when the lid is opened. Therefore, the working environment in the room is deteriorated, and more than the required amount of the medicine is consumed. This problem has a great influence when working in a small room, and is a serious problem particularly when performing experiments in the shuttle in space. Therefore, in order to minimize the amount of leakage into the room, the required amount of the drug is accurately calculated and the work is performed with the minimum required amount.

As described above, in the conventional technique, it is necessary to have skill in the operation of scattering and injecting the drug into the room, and since the minimum amount of drug is used, the purpose of drug injection such as immobilization cannot be achieved. There are problems such as occurrence of. The present invention has been made in view of the state of the art of the prior art described above, and provides a plant cell culturing apparatus which is easy to operate and which can inject a drug without fear of scattering of the drug.

[0005]

Means for Solving the Problems The present invention is (1) a plant cell culturing apparatus comprising a culture container for accommodating a medium and an upper lid of the container, having a vent hole, which is hermetically sealed during culturing and, when necessary, via a septum seal. A plant cell culturing apparatus having an injection port to which a drug injection syringe can be attached, and (2) a plant cell culturing apparatus having a vent and a culture container and a container upper lid for storing a medium. Therefore, it is possible to attach an injection port that is tightly plugged during culturing and can be attached with a syringe for drug injection through the septum seal when necessary, and a dust bag that is tightly plugged during culturing and communicates with the container through the septum seal when needed. An apparatus for cultivating plant cells, comprising: a dust collecting bag attachment port.

[0006]

1 is a conceptual diagram showing an example of an apparatus and an operating method thereof according to the first aspect of the present invention. This device comprises a culture container 1 and a container upper lid 2, and the upper lid is provided with an injection port 3 and a vent hole 4. Cultivation of plant cells is performed with the injection port 3 sealed and the vents 4 open. When the culture is stopped, the ventilation hole 4 is sealed with a sealing pad 5 or the like. Then, the cap of the injection port 3 is removed, the air bleeding syringe 8 is attached, and part of the gas in the device is sucked and removed (FIG. 1).
(A)). Next, the medicine is injected by replacing the medicine injection syringe 9 (FIG. 1 (b)). In this case, since the gas inside the container is partially removed, the drug can be injected smoothly.

FIG. 2 is a conceptual diagram showing an example of an apparatus and a method of operating the same according to the second aspect of the present invention. This apparatus comprises a culture container 1 and a container upper lid 2, and the upper lid is provided with an injection port 3, a dust bag attachment port 6 and a ventilation hole 4. Cultivation of plant cells is performed in a state where the injection port 3 and the dust bag attachment port 6 are tightly plugged and the vent hole 4 is opened. When the culture is stopped, the ventilation hole 4 is sealed with a sealing pad 5 or the like. Next, the caps of the dust bag attachment port 6 and the injection port 3 are removed, and the dust bag 7 and the drug injection syringe 9 are attached (FIG. 2A). When the medicine is injected from the medicine injection syringe 9 in this state, the gas containing the medicine vapor flows out into the dust bag 7, whereby the medicine is smoothly injected (FIG. 2B). In this device, if the injection port 3 and the dust bag attachment port 6 are provided at positions symmetrical with respect to the center axis of the culture device, degassing will be carried out more smoothly due to internal convection. It should be noted that also in the apparatus of FIG. 2, the dust bag attachment port 6 may be left closed, and the drug may be injected after degassing from the injection port 3 as in the case of the apparatus of FIG. .

[0008]

The present invention will be described in more detail with reference to the following examples. 3 (a) and 3 (b) are schematic views showing an example of the plant cell culture device of the present invention, and FIG. 4 is a structure of the injection port 3 and the dust bag attachment port 6 and dust collection of the device of FIG. FIG. 5 is an explanatory diagram showing how the bag 7 and the drug injection syringe 9 are attached, and FIG. 5 is an explanatory diagram showing a state of the drug injection work. This device comprises a culture vessel 1 and a vessel top lid 2 each having a substantially circular cross section, and the top lid has a vent hole 4 in the center.
However, the injection port 3 is provided at a position close to the peripheral portion, and the dust bag attachment port 6 is provided at a position substantially symmetrical to the injection port 3 and the central axis of the apparatus. During the cell culture, only air flows in and out from the ventilation hole 4 through the ventilation membrane 18 such as a Teflon filter supported by the retainer 21. The culture container 1 and the container upper lid 2 are fastened with four screws 12 and sealed with a seal ring 11.

For culturing plant cells, first, a medium such as agar is put on the honeycomb sheet 10 in the culture container 1 to put the sample plant cells. Attach the container top lid 2 and tighten with screws 12. This seals with the seal ring 11. During the culture, the drug injection port 3 is tightly plugged with the cap 15 via the septum seal 13, and the dust bag attachment port 6 is also tightly plugged with the cap 16, packing 19 and septum seal retainer 20 via the septum seal 14. . The air required for the culture is supplied through a filter attached to the ventilation hole 4. The septum seal can maintain airtightness in the normal state, and the injection needle (needle)
The airtightness is maintained by the expansion of the septum after the needle is pulled out. When the cell culture is completed, a drug such as glutaraldehyde is injected to fix the cells. Prior to injecting the drug, a sealing pad 5 is attached to the ventilation hole 4 and sealed by tape or the like. As a result, the inside of the culture device is completely shielded from the outside air.

Next, as shown in FIG. 4, the cap 16 of the dust bag mounting port 6 is removed, and the dust bag 7 in a state in which the air inside is removed and the needle 17 at the tip of the dust bag 7 are attached to the dust bag mounting port 6. The septum seal 14 is penetrated and attached. Then the cap 15 of the injection port 3
Then, the drug injection syringe 9 having a needle at its tip is inserted into the septum seal 13 and attached. In this state (state of FIG. 5), the drug in the drug injection syringe 9 is injected into the culture device. Due to the pressure of the injection, a part of the vaporized components of the drug such as glutaraldehyde is stored in the dust bag 7 together with the air in the device. After the injection is completed, the drug injection syringe 9
And the dust bag 7 is removed, and the caps 15 and 16 are tightly plugged to complete the immobilization. Since both the drug injection syringe 9 and the dust bag 7 are attached via the septum seals 13 and 14, they are sealed by the septum seal even when they are removed, so that the vaporized drug should be jetted and scattered into the room. There is no.

Here, an example in which a dust bag is used has been described, but the structure of the injection port portion may be substantially the same even in the case of a culture device that does not have a dust bag attachment port. The working method is to attach the air bleeding syringe before attaching the drug injecting syringe and only partially remove the gas inside, and the operation of attaching and detaching the air bleeding syringe is the same as in the case of the drug injecting syringe. Alternatively, a device having a dust bag attachment port may be used to inject the drug by a method using an air-bleeding syringe without using the dust bag.

[0012] The drug injection amount depends on the type of culture species, the culture scale,
Although it depends on the culture conditions, the drug used, the purpose of injecting the drug, etc., it is an example in which cell culture is carried out by a device having a diameter of 100 mm and a height of 30 mm and immobilized by glutaraldehyde, and the drug injection amount is about 20 ml. . The device of the present invention is convenient for fixing cells by injecting glutaraldehyde, but it is of course effective for injecting various drugs during or after the progress of culture.

[0013]

The plant cell culture device of the present invention has the following advantages. (1) Since an injection port that can attach an air-bleeding syringe or a drug injection syringe via a septum seal is provided, the vent hole is closed, and a part of the gas inside is removed by the air-bleeding syringe before the drug is dispensed. Since the injection can be performed, the injection work can be performed smoothly and the drug is not scattered. (2) Since the injection port to which the medicine injection syringe can be attached via the septum seal and the dust bag attachment port to which the dust bag can be attached via the septum seal are provided, the vent hole is closed. Since it is possible to inject the drug while accommodating a part of the internal gas containing the vaporized drug in the dust bag, the injection work can be performed smoothly and the drug is not scattered. (3) Since the preparation for injecting a drug is completed simply by inserting a syringe or a needle of a dust bag into the septum seal without disassembling the cell culture device, the number of working steps can be reduced to 1/3 or less. (4) Since the medicine does not go out of the device at all or only flows out into the dust bag, the necessary amount of the medicine can be easily calculated, and the work such as immobilization can be surely performed.
Waste of drug consumption can be eliminated.

[Brief description of drawings]

FIG. 1 is a conceptual diagram showing an example of an apparatus and an operating method thereof according to a first invention of the present invention.

FIG. 2 is a conceptual diagram showing an example of an apparatus and an operating method thereof according to a second aspect of the invention.

FIG. 3 is a schematic view showing an example of a plant cell culture device of the present invention.

4 is an explanatory view showing the structure of an injection port 3 and a dust bag attachment port 6 of the apparatus of FIG. 3 and the attachment situation of a dust bag 7 and a drug injection syringe 9;

FIG. 5 is an explanatory view showing a state of a drug injection work.

FIG. 6 is a schematic view showing an example of a conventional plant cell culture device.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Muneo Takaoki 1-1-1, Wadasaki-cho, Hyogo-ku, Kobe-shi, Hyogo Mitsubishi Heavy Industries, Ltd. Kobe Shipyard (72) Inventor Yasuki Kamo Hyogo, Kobe-shi, Hyogo 1-1-1, Wadasaki-cho, Tokyo Mitsubishi Heavy Industries Ltd. Kobe Shipyard

Claims (2)

[Claims] 1. A plant cell culture device comprising a culture container for accommodating a medium and a container upper lid, and having a vent hole, wherein a syringe for drug injection can be attached through a septum seal when the culture container is tightly plugged during the culture. A plant cell culture device having a port. 2. A plant cell culture apparatus comprising a culture container for accommodating a medium and a container top lid, and having a vent hole, wherein the syringe is tightly closed during the culture and a drug injection syringe can be attached via a septum seal when necessary. A plant cell culturing apparatus comprising: a port; and a dust bag attachment port to which a dust bag that is tightly plugged during culture and communicates with a container via a septum seal when necessary can be attached.