JP3546077B2 - Plant cell culture equipment - Google Patents

Description

[0001]
[Industrial applications]
The present invention relates to a plant cell culture apparatus suitable for cell culture in a small room or in a microgravity environment and capable of injecting a drug without fear of environmental pollution.
[0002]
[Prior art]
As a device for performing a plant cell culture test, a Petri dish type culture device as shown in FIGS. 6A and 6B is used. This apparatus is composed of a culture container 1 and a container upper lid 2. A culture medium such as an agar medium is provided on a base material such as a honeycomb sheet 10 in the culture container 1, and a plant cell is inoculated in the culture medium to perform a culture experiment. . During the culturing, outside air is taken in from the vent hole 4 provided in the container top lid 2 through a permeable filter (vent membrane 18) such as a Teflon filter, whereby the cells are respired and cultured.
When the culture for a predetermined time is completed, the cells are taken out and various tests are performed.For example, when the culture place is separated from the test place, such as when performing a culture test in a shuttle in space, the test is performed immediately after the culture is completed. In many cases, the test cannot be performed or time is required before the test for some reason. In such a case, if left as it is, cultivation further proceeds, and accurate data cannot be obtained. Therefore, a method of injecting a drug such as glutaraldehyde into a container to stop the cultivation has been adopted. This work is called fixed work.
[0003]
[Problems to be solved by the invention]
When injecting the drug into such a culture device in a laboratory on the ground or the like, the lid of the petri dish is removed and injected. However, in this case, the toxic agent may leak into the room because the device is not sealed during the injection operation, and the contents may be scattered when the lid is opened in a microgravity space such as space. As a result, the working environment in the room deteriorates, and more medicine is consumed than necessary. This problem has a great effect when working in a small room, especially when performing experiments inside a shuttle in outer space. Therefore, in order to minimize the amount of leakage into the room, the required amount of the medicine is accurately calculated, and the operation is performed with the minimum required amount.
[0004]
As described above, in the conventional technique, the medicine is scattered into the room, skill is required for the injection operation, and the purpose of the injection of the drug such as immobilization may not be achieved because the minimum amount of the drug is used. There is a problem.
The present invention has been made in view of the state of the art of the prior art, and provides a plant cell culture device that is easy to operate and can inject a drug without fear of scattering of the drug.
[0005]
[Means for Solving the Problems]
The present invention is made from a culture vessel, and vessel head for accommodating a culture ground, a plant cell culture device having a vent hole, the injection port can be attached to the drug injection syringe via the septum seal at the time of need is sealed in culture And a dust bag attaching port to which a dust bag that is sealed during culture and can communicate with the container via a septum seal when necessary can be attached.
[0006]
[Action]
FIG. 1 is a conceptual diagram showing an example of an apparatus according to a reference example of the present invention and an operation method thereof. This apparatus comprises a culture vessel 1 and a vessel top lid 2, which is provided with an injection port 3 and a vent 4. The cultivation of the plant cells is performed with the injection port 3 sealed and the vent hole 4 opened. When the culture is stopped, the vent hole 4 is sealed with a sealing pad 5 or the like. Next, the cap of the injection port 3 is removed, the air vent syringe 8 is attached, and a part of the gas in the device is removed by suction (FIG. 1A). Next, the medicine is injected by replacing the medicine injection syringe 9 (FIG. 1B). In this case, since the gas inside the container is partially removed, the injection of the medicine is performed smoothly.
[0007]
Figure 2 is a conceptual diagram showing an example of a device and its method of operation according to the onset bright. This apparatus comprises a culture vessel 1 and a vessel top lid 2, which is provided with an injection port 3, a dust bag attachment port 6, and a vent 4. The cultivation of the plant cells is performed in a state where the injection port 3 and the dust bag attachment port 6 are sealed and the vent hole 4 is opened. When the culture is stopped, the vent 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 medicine injection syringe 9 are attached, respectively (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 collection 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 central axis of the culture device, gas can be more smoothly removed by internal convection. In the apparatus of FIG. 2, the dust bag attachment port 6 may be left closed, and the medicine may be injected after degassing from the injection port 3 as in the apparatus of FIG. .
[0008]
【Example】
Hereinafter, the present invention will be described more specifically with reference to examples.
3 (a) and 3 (b) are schematic diagrams showing an example of the plant cell culture device of the present invention, and FIG. 4 is a diagram showing the structure of the injection port 3 and the dust bag attachment port 6 of the device of FIG. FIG. 5 is an explanatory view showing a state of attachment of the bag 7 and the drug injection syringe 9, and FIG. 5 is an explanatory view showing a state of a drug injection operation.
This device comprises a culture vessel 1 and a vessel top lid 2 having a substantially circular cross section. The top lid has a vent hole 4 at the center, and an injection port 3 at a position close to the periphery and the injection port 3 and the central axis of the apparatus. A dust bag attachment port 6 is provided at a substantially symmetric position. During the culturing of the cells, only air enters and exits from the vent hole 4 through the gas permeable membrane 18 such as a Teflon filter supported by the presser 21. The culture vessel 1 and the vessel top lid 2 are fastened with four screws 12 and sealed with a seal ring 11.
[0009]
For cultivation of plant cells, first, a medium such as agar is put on the honeycomb sheet 10 in the culture vessel 1 and plant cells of a sample are put. Attach the container upper lid 2 and tighten with the screw 12. This seals with the seal ring 11. During the culture, the drug injection port 3 is sealed with a cap 15 via a septum seal 13, and the dust bag attachment port 6 is also sealed with a cap 16, a packing 19 and a septum seal holder 20 via a 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 a normal state, can insert an injection needle (needle), and can maintain airtightness by expanding the septum after extracting the needle.
When the cell culture is completed, a drug such as glutaraldehyde is injected to fix the cells. Prior to the injection of the drug, a sealing pad 5 is attached to the ventilation hole 4 and sealed by a method such as sealing with a tape. Thereby, the inside of the culture device is completely shut off from the outside air.
[0010]
Next, as shown in FIG. 4, the cap 16 of the dust bag attachment port 6 is removed, and the dust bag 7 in a state where the air is removed from the dust bag attachment port 6 is connected to the septum seal 14 of the dust bag attachment port 6 with the needle 17 at the tip. And attach it. Next, the cap 15 of the injection port 3 is removed, and the medicine injection syringe 9 having a needle at the tip is inserted into the septum seal 13 and mounted. In this state (the 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 component of the drug such as glutaraldehyde is stored in the dust collection bag 7 together with the air in the device.
After the injection is completed, the drug injection syringe 9 and the dust collection bag 7 are removed, and sealed with the caps 15 and 16 to complete the immobilization. Since both the drug injection syringe 9 and the dust bag 7 are mounted via the septum seals 13 and 14, they are sealed by the septum seal even when they are removed, so that the vaporized drug spouts into the room and scatters. There is no.
[0011]
Here, an example in which the dust bag of the present invention is used has been described. However, the structure of the injection port portion may be substantially the same even in the case of a culture device (FIG. 1) having no dust bag attachment port. The working method is also the same as in the case of the drug injection syringe, except that the air bleeding syringe is attached and only a part of the gas inside is evacuated before the drug injection syringe is attached. Alternatively, the device of the present invention having a dust bag attachment port may be used to inject medicine by using a bleeding syringe without using a dust bag.
[0012]
The amount of drug to be injected varies depending on the type of culture species, culture scale, culture conditions, used drug, purpose of injecting the drug, and the like. Cell culture is performed using a device having a diameter of 100 mm and a height of 30 mm, and immobilization with glutaraldehyde. In the example, the drug injection volume is on the order of about 20 milliliters.
The device of the present invention is convenient for fixing cells by injecting glutaraldehyde. However, it is needless to say that the device of the present invention is also effective for injecting various drugs during or after culturing.
[0013]
【The invention's effect】
The plant cell culture apparatus of the present invention has the following advantages.
(1) An injection port to which an air bleeding syringe or a medicine injecting syringe can be attached via a septum seal is provided. With the vent hole closed, a part of the gas inside the air bleeding syringe is removed, and then the medicine is removed. Can be injected, the injection operation can be performed smoothly, and there is no scattering of the medicine.
(2) Since an injection port to which a medicine injection syringe can be attached via a septum seal and a dust bag attachment port to which a dust bag can be attached via a septum seal are provided, the vent hole is closed. The medicine can be injected while a part of the internal gas containing the vaporized medicine is stored in the dust bag, so that the injection operation can be performed smoothly and the medicine does not scatter.
(3) Since the preparation for injecting the drug is completed only by inserting the syringe or the needle of the dust bag into the septum seal without disassembling the cell culture device, the number of man-hours can be reduced to 1/3 or less.
(4) Since the medicine does not come out of the apparatus at all or only flows out into the dust bag, it is easy to calculate the required amount of the medicine, and the operation such as immobilization can be reliably performed, and the consumption of the medicine can be increased. Can be eliminated.
[Brief description of the drawings]
FIG. 1 is a conceptual diagram showing an example of an apparatus according to a reference example of the present invention and an operation method thereof.
Figure 2 is a conceptual diagram showing an example of a device and its method of operation according to the onset bright.
FIG. 3 is a schematic diagram showing one example of a plant cell culture device of the present invention.
FIG. 4 is an explanatory view showing the structure of an injection port 3 and a dust bag attaching port 6 of the apparatus shown in FIG. 3, and the attachment state of a dust bag 7 and a medicine injection syringe 9;
FIG. 5 is an explanatory view showing a state of a medicine injection operation.
FIG. 6 is a schematic view showing an example of a conventional plant cell culture device.

Claims (1)

A plant cell culturing apparatus comprising a culture container for storing a culture medium and a container top lid and having an air hole, wherein an injection port capable of being attached with a syringe for drug injection via a septum seal when necessary during culturing, And a dust bag attachment port to which a dust bag that can be attached to the container via a septum seal when necessary can be attached .

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