JPH06327461A - Tissue culture learning kit - Google Patents

Abstract

PURPOSE:To obtain the subject kit enabling experiments to be conducted easily and assuredly, by providing a partition wall means to divide the inner space of a sterilized bag with fasteners into plural subspaces and by putting a sterilized medium, etc., in one of the subspaces. CONSTITUTION:A bag 2 capable of dividing its inner space with fasteners 21, 23 and of blocking the resultant subspaces one another is sterilized and served as a clean bench. One of such subspaces is housed with capped culture equipment 30, 31, a high-molecular water absorber 32, a powdery medium 33, water 34 and a forceps 35. The medium and the others should have also been sterilized. With this kit, anybody can conduct experiments easily.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a tissue culture learning kit which enables easy cell culture experiments.

[0002]

2. Description of the Related Art Conventionally, there has been a demand for actually culturing cell tissues in school classes and the like. Several types of tissue culture learning kits are commercially available to meet such demands.

For example, a carrot culture kit contains carrot callus, a growth medium, a differentiation medium, and a rooting medium. The learner transplanted these callus to each medium in a clean bench or a sterile box, and as a basis of aseptic operation or induction of adventitious embryos from callus, induction of adventitious roots, proliferation of callus,
You can learn the actual tissue culture.

[0004]

However, the above-mentioned conventional tissue culture learning kit is premised on that the learner is equipped with equipment such as a clean bench. Therefore, not everyone was able to perform tissue culture experiments.

Further, there is a problem that the culture medium is generally deteriorated about one month after its preparation and the culture efficiency is deteriorated, so that the culture medium cannot be reserved. For example, if the MS medium is left for a long period of time, calcium ions and phosphate ions may react with each other to form a sparingly soluble salt, which may cause precipitation. As a result, there is a problem that the ionic composition of the medium changes.

[0006] In addition, sterilization with an autoclave (for example, 1
There was also a problem that the PH value was lowered when it was carried out at 21 ° C. for 15 minutes (Note: it is said that this is because sucrose is denatured by heat). In addition to this, the medium also contains various amino acids and vitamins, and there is a risk that these may chemically change in the autoclave or combine with various ions.

An object of the present invention is to provide a tissue culture learning kit that enables anyone to easily and surely perform a tissue culture experiment.

[0008]

Means for Solving the Problems The present invention has been made in order to achieve the above object, and in one aspect thereof, is composed of a material capable of blocking the entrance and exit of at least microorganisms that may affect culture experiments, And, a bag whose inner space is sterilized, a closing means for closing the opening of the bag in an airtight manner, and a partition wall capable of partitioning the inner space of the bag in at least two independent chambers in an airtight manner There is provided a tissue culture learning kit comprising a means and a sterilized medium stored in any one of the above rooms.

The culture medium comprises water and a nutrient component required by a tissue to be cultured, and comprises a packaging container for storing the water and the nutrient component, It is preferable that the water and the nutrient component are stored in the packaging container in separate states. Moreover,
The water and the nutrient component are preferably packaged in the packaging container at a volume ratio necessary for forming a medium having a concentration suitable for culture.

According to another aspect of the present invention, a bag made of a material capable of blocking entry and exit of microorganisms which may affect a culture experiment, and a closure capable of closing the opening of the bag in an airtight manner. There is provided a simplified clean bench, characterized in that it comprises a means and a partition means capable of partitioning the inner space of the bag into at least two independent chambers while maintaining airtightness. It

[0011]

The bag whose inside can be airtightly divided by the partition means and the closing means can be used as a clean bench for conducting a culture experiment. This eliminates the need to prepare special equipment in advance.

In this case, anyone can easily carry out the experiment by preparing the required medium and the like in a bag in a sterilized state. Furthermore, the medium can withstand long-term storage by keeping water and nutrient components separated.

[0013]

An embodiment of the present invention will be described with reference to the drawings.

The tissue culture learning kit of this example is shown in FIG.

The tissue culture learning kit 1 comprises incubators 30 and 31 with a lid, a polymeric water absorber 32, and a powder medium 33.
, Water 34, tweezers 35, and a bag 2 containing these.

The bag 2 is used not only for accommodating the incubator 30 and the like, but also for carrying out a tissue culture experiment.
It is used as a bench. Therefore, the bag 2
Is composed of a material capable of blocking gases and microorganisms (virus, bacteria, etc.) that may affect experiments. In this embodiment, ordinary vinyl is used as the material. However, for example, when examining the influence of ultraviolet rays, a material that blocks ultraviolet rays may be used. Fasteners 21 and 23 capable of blocking the flow of gas and miscellaneous bacteria are provided in the opening and inside of the bag 2. Therefore, when the fasteners 21 and 23 are closed, the first aseptic chamber 22 and the second aseptic chamber 24 are formed inside the bag 2.

As will be described later, during the experiment, the incubator 3
With 0 and the like kept in the bag 2, the operation is performed by holding these from the outside of the bag 2. Therefore, the bag 2 is preferably made of a relatively thin and flexible material. The means for closing the opening of the bag 2 and the means for partitioning the inside of the bag 2 are not limited to fasteners. For example, it may be opened and closed by disposing a lid member having adhesiveness. The bag 2 may be completely sealed at the outer position of the fastener 21 by thermocompression bonding and the sealed portion may be broken at the start of the experiment. In this way, there is less risk that the fastener 21 will open during the distribution process and the sterilized state will be broken.

The incubator 31 can be housed in the incubator 30. The lids of both incubators 30 and 31 are airtight enough to prevent bacteria from entering.

The water 33 and the powder medium 34 are put in a suitable container in a sterilized state and stored in the second aseptic chamber 24 of the bag 2 (the inside of the bag 2 is also sterilized beforehand). Needless to say). The composition of the nutritional components of the powder medium 34 is adjusted according to the plant to be cultivated. The amount of water 33 contained in the kit is preferably such that when the powder medium 34 contained in the kit is dissolved, a medium having an appropriate concentration can be obtained. Assuming that the kit is used multiple times, water 33, powder medium 34
May be put in multiple times, but even in that case, the water 33 and the powder medium 34 should be packaged separately for each amount necessary to form a medium having an appropriate concentration for easy experimental operation. Is preferred. For example, when an MS medium manufactured by Nippon Pharmaceutical Co., Ltd. is used as the powder medium, it is prepared in a ratio of 4.9 g of the powder medium drug to 1000 g of water (Note: the powder used in this example. The medium does not contain sugar, so add sucrose, etc.).

Furthermore, water 33 contained in one sachet
And the amount of liquid medium produced by mixing the powder medium 34 contained in one sachet are
It is determined according to the water absorption capacity of 2 and the volume of the incubator 31.

In this example, as the polymer water absorber, trade names "Aqualic" and "Acryhop" manufactured by Nippon Shokubai Co., Ltd. are used.

In addition to the above, tools and chemicals may be added to the kit as needed.

The operation procedure of the tissue culture experiment will be described.

Preparation of medium The powder medium 34 is put in the water 33 and shaken well. This allows
It is possible to produce a liquid medium having an appropriate concentration. Next, the melted aqueous solution (liquid medium) of the powder medium 34 is put into the container 31. As a result, the aqueous solution (liquid medium) is absorbed by the polymer water absorber 32 and becomes a solid medium. As a matter of course, this work is performed with the container 31 and the like kept in the sterile room 24. That is, the work is performed from the top of the bag 2 with a container or the like containing water 33 or the like.

The tissue C to be transplanted and cultured includes an inner container 50 and an outer container 52.
It is premised that the container 5 is contained in the double container 5 in advance. It goes without saying that both the inner container 50 and the outer container 52 are provided with lids sufficient to block the entry of bacteria and the like.

The outer container 52 is sterilized by wiping it with 70% ethanol or the like. Then, the fastener 31 is opened, the tissue C together with the container 5 is placed in the first sterile chamber 22, and the fastener 31 is closed again. When this operation is performed, the fastener 23 must be closed in order to prevent bacteria and the like from entering the second sterile chamber 24. It is important to perform the work quickly so as to minimize the invasion of bacteria and the like in the outside air. It should be noted that, together with the container 5, a gauze or the like impregnated with ethanol is also put therein, and after the fastener 31 is closed, the outer surface of the outer container 52 and the first sterile chamber are sterilized again. You can go.

All the subsequent work is performed from the outside of the bag 2, and the container 50 and the like are not directly exposed to the outside air.

First, the outer container 52 is opened, and the inner container 50 is taken out and transferred to the second aseptic chamber 24. And the inner container 50
Then, open the container 31 and use the forceps 35 to remove the tissue C.
Is transferred from the inner container 50 into the container 31. Then the container 31
The lid is closed and the container 31 is placed in the container 30.

Finally, with the container 30 covered, the bag 2
The container 30 is taken out from the container and the transplanting work is completed.

When subculture is performed, a new tissue culture kit is used in the same manner. In this case, the containers 30, 31
Corresponds to the inner container 50 and the outer container 52 in the above description.

Incubator 30 for a plurality of times in the second sterile chamber 24
If you prepare such as one tissue culture learning kit,
Continued experiments can be carried out.

As described above, the tissue culture learning kit of the present invention allows the user to experience a tissue culture experiment without preparing special equipment such as a clean bench. In addition, since the medium is divided into water, a polymeric water absorber, and a powder medium, the medium does not deteriorate even if the kit is sterilized with an autoclave (Note: the entire kit, that is, a bag. It is difficult to avoid heating the medium, since sterilization of the whole of 2 will be performed at the final stage of the kit production.). In addition, it can withstand long-term storage. Therefore, it is possible to reserve the tissue culture learning kit.

Further, since the water 33 and the powder medium 34 are separately packaged according to the quantity required for producing a medium having an appropriate concentration, troublesome weighing operation is unnecessary. Further, the amounts of the water 33 and the powder medium 34 contained in each package are determined according to the water-absorbable amount of the polymer water-absorbing body 32 and the volume of the incubator 31, so that the produced liquid medium remains. There is no waste. In particular, in the tissue culture learning kit of this example, since the experiment operation is performed from the top of the bag 2, it is particularly important to take such a measure for simplifying the operation in the case of an actual product. is there. In addition to selling as a tissue culture learning kit, it is also possible to separately sell water and a powder medium that are packaged in such a volume ratio as a "medium".

Moreover, by preparing all the substances such as water that can be easily obtained in the kit in advance, regardless of the difference of the experimenter and the difference of the experiment environment,
Tissue culture can be surely successful. Ensuring such certainty is also important in assuring the reliability of commercial products. For example, if the experimenter is a high school student, it is easy to prepare distilled water, but if the experimenter is an elementary school student, it is difficult to desire to prepare distilled water. Even if the instruction to use distilled water is given in the instruction manual, tap water is often used as it is. The quality of the tap water (particularly the concentration of the sterilizing agent) varies greatly depending on the region, and the difference in quality may affect the result of the culture.

In the above embodiment, the incubators were used in a double stack. However, if the bacteria are completely blocked by the lid of the incubator and the sterilization is sufficiently performed with ethanol or the like, it is not always necessary to make the cells double. Alternatively, on the contrary, when it is desired to more reliably block bacteria and the like (for example, when the tissue culture learning kit is premised on tissue culture of a plant that is extremely susceptible to bacteria), the bag 2 should be further It may be divided into a large number of sterile rooms. For example, as shown in FIG. 3, a fastener 25 is further provided in the bag 2 ′ to form a third sterile chamber 26 further on the inner side than the second sterile chamber 24. And incubator 3
0, etc., if put in the third sterile room 26,
Bacterial blockage becomes more reliable. From the first sterile room 22 to the second
To the sterile room 24, and from the second sterile room 24 to the third sterile room 2
It is needless to say that it is preferable to sequentially perform sterilization by using ethanol or the like when moving the tissue to 6 as in the case described above. Further, when it is desired to enable a plurality of types of experiments with the same kit, or when it includes a plurality of operations that are preferably performed in mutually isolated places,
As shown in Fig. 4, the second aseptic chamber 24 in the bag 2 "may be divided into two (in the figure, the divided second aseptic chamber is indicated by reference numerals 24a and 24b). By changing the configuration in various ways, the bag 2 can be widely used as a clean bench for purposes other than tissue culture.

Since the bag 2 can be folded and made small, even if the bag 2 itself is made large, it can be folded and placed in the autoclave. Therefore, it is naturally possible to make the bag 2 larger by providing the support pillars or the like (Note: in the above-described embodiment, the support pillars are not provided because the bag 2 is relatively small). It is preferable that the columns have a foldable structure.

An example of the columns is shown in FIG. The column 9 in this figure has a structure capable of expanding and contracting like a rod antenna. In this case, the connection with the other support column 9 is performed by inserting the other support column 9 into the rings 900 at the upper and lower ends. In addition, it is preferable that the support column 9 be detachable from the bag 2 without opening and closing the fasteners 21 and 23 of the bag 2. For example, a ring through which the pillar 9 is passed may be provided on the side of the bag. With such a structure, even if the bag is enlarged, it can be folded and auto-clad.
The sterilization can be performed by using a bur. Since the column is removable, it does not hinder the sterilization work.

Further, as shown in FIG. 6, an air chamber 65 independent of the sterile chamber may be provided at the side of the bag 100, and the air chamber 65 may be used as a support. Although only the bag is shown in FIG. 6, it goes without saying that, if necessary, the equipment and medium necessary for tissue culture are put in the second sterile chamber 24 in a sterilized state.

In this case, if the air in the air chamber 65 is evacuated, it can be folded as it is to make it smaller, and it can be put in an autoclave or the like. Further, when used, if air is introduced into the air chamber 65, the sterile chambers 22 and 24 in the bag 100 can be supported as an air column. Further, since the sterilization filter 70 is provided, the outside air can flow into the aseptic chambers 22 and 24 in an aseptic state. Therefore, even if air is introduced into the air chamber 65, the aseptic chamber does not have a negative pressure and the aseptic chambers 22 and 24 are not inflated sufficiently. It goes without saying that the air injected into the air chamber 65 does not have to be sterilized. Although not shown in this figure, if a part of the wall surfaces of the aseptic chambers 22 and 24 is formed into a glove shape, an experiment operation can be performed by putting a hand in the glove portion. Will be easier. When the support is configured as an air chamber in this way, the support is extremely unlikely to damage the bag 2 ′ and break the aseptic condition.
Easy to handle. Further, since the rod-shaped support is not included, it is effective in handling the product and occupying the area in the store when it is sold. Also, the manufacturing cost can be reduced. In addition, it is possible to sell only the above-mentioned bag 100 etc. as a simple clean bench.

[0040]

By using the tissue culture learning kit of the present invention, the basics of tissue culture can be experienced without preparing a special device. In addition, since the culture medium is produced when the experiment is performed, the tissue culture learning kit can be reserved. Further, since the bag is used as a clean bench, it occupies a small area and is convenient when displayed in a store.

[Brief description of drawings]

FIG. 1 is a view showing a tissue culture learning kit which is an embodiment of the present invention.

FIG. 2 is a view showing a container in which a tissue to be cultured is previously placed.

FIG. 3 is a diagram showing an example of a bag 2 ′ having different internal division methods.

FIG. 4 is a diagram showing an example of a bag 2 ″ having different internal division methods.

FIG. 5 is a diagram showing an example of a column.

FIG. 6 is a view showing a bag 100.

[Explanation of symbols]

1: Tissue culture learning kit, 2: Bag, 5: Container, 2
1: Fastener, 22: First sterile room, 23: Fastener, 24: Second sterile room, 30: Incubator, 3
1: Incubator, 32: High-molecular water absorber, 33: Powder medium, 34: Water, 35: Tweezers, 50: Inner container,
52: Outer container

Claims (4)

[Claims] 1. A bag which is made of a material capable of blocking entry and exit of microorganisms which may affect culture experiments, and whose inner space is sterilized, and a closing means for closing the opening of the bag in an airtight manner. And a partition means capable of partitioning the inner space of the bag into at least two independent chambers while maintaining airtightness, and a sterilized medium stored in any one of the chambers, Tissue culture learning kit. 2. The medium comprises water and a nutritional component required by a tissue to be cultured, and a packaging container for storing the water and the nutritional component is provided. The tissue culture learning kit according to claim 1, further comprising: the water and the nutrient component stored in the packaging container in a separated state. 3. The water and the nutrient component are packaged in the packaging container at a volume ratio necessary for forming a medium having a concentration suitable for culture. The described tissue culture learning kit. 4. A bag made of a material capable of blocking entry and exit of microorganisms that may affect culture experiments, a closing means capable of closing the opening of the bag in an airtight manner, and the inside of the bag. A simple clean bench comprising: a partition means capable of partitioning the space into at least two independent rooms that are kept airtight if necessary.