JPH11342179A - Sealing method and sealing device for communication channel in multiple chamber freezing bag - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method and device for preventing a breakage accident in thawing a multiple chamber freezing bag. SOLUTION: This sealing device 1 includes a table 11 for a multiple chamber freezing bag 2 to be placed horizontally, a means (a cover 12 and a lock 13) for regulating quantity of cell flotation liquid contained in the multiple chamber freezing bag 2 placed on the table 11, and a sealing means (an impulse sealer 15) for sealing a communication channel in the multiple chamber freezing bag 2.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method and a device for sealing a channel connecting a plurality of chambers of a multi-compartment freezing bag. Specifically, after filling the cell suspension in a multi-compartment freezing bag with multiple compartments, before freezing,
The present invention relates to a method and an apparatus for previously sealing a channel connecting a plurality of chambers in preparation for partial thawing.

[0002]

2. Description of the Related Art Conventionally, as a method for preserving a cell suspension derived from a living body such as a blood cell for a long period of time, a method has been adopted in which a cell suspension is stored in a flat type freezing bag having two closely adhered surfaces and cryopreserved. I have. In recent years, water crystals in cells (ice)
It has been shown that the behavior of the cell affects the damage of the cell membrane and consequently the cell survival. In addition, it was found that the behavior of water crystals in the cells depends on the cooling rate of the cell suspension. Therefore, when the cell suspension is stored in a freezing bag and cryopreserved, in order to maintain the viability of the cells during freezing and after thawing, the water in the cells becomes fine water crystals that do not damage the cell membrane from the start of freezing. The cooling rate must be rigorously adjusted to a stable temperature. Then, in order to strictly adjust the cooling rate of the cell suspension, it is necessary to make the thickness of the cell suspension contained in the freezing bag uniform over the entire surface of the freezing bag.

However, when a cell suspension is stored in a conventional flat frozen bag, the bag shape after storage changes greatly from a plate shape to a pillow shape according to the storage amount. Since the range is quite large and the thickness of the liquid over the entire surface of the bag after containing the cell suspension is small at the end of the bag and large at the center, the temperature of the cell suspension during cooling can be rapidly cooled. Low at the center and high at the center where the cooling is delayed. Therefore, in the flat type freezing bag, it is impossible to freeze the contained cell suspension at a uniform speed, no matter how strictly the cooling temperature is controlled. The greater the capacity, the more intense. Therefore, recently, in order to solve such a problem, a so-called three-dimensional freezing bag having a flat rectangular chamber has been adopted.

[0004] This three-dimensional freezing bag can easily freeze the entire cell suspension at a uniform speed because the thickness of the cell suspension becomes uniform over the entire bag when the cell suspension is stored therein. It is. However, even in the case of using a three-dimensional freezing bag, if the volume of the cell suspension to be stored exceeds the specified volume, water may expand due to freezing and damage the bag, and If the bag is frozen without holding the bag horizontally, the cell suspension will move to a lower level and become swollen, which will not only freeze the entire cell suspension at a uniform rate, but also cause damage to the bag. Sometimes. For this reason, recently, a three-dimensional double-chamber frozen bag is housed in a flat rectangular container (canister) so that the bag can be frozen at a uniform speed without holding the bag in a horizontal state and the bag can be prevented from being damaged. Then, a method of freezing while keeping the thickness of the cell suspension constant has been adopted.

[0005] On the other hand, in clinical application of the cell suspension, it is required to use the same cell suspension several times, for example, to thaw only a part of the same cell suspension to proliferate the cells. And for that,
Recently, three-dimensional double-chamber freezing bags have been adopted. However, when the cell suspension is stored and frozen in such a multi-chamber freezing bag, communication between the chambers is performed after storing the cell suspension so that only a part of the cell suspension can be used separately. The channels are sealed and frozen, but during freezing, the smaller chamber of the multi-chamber freezing bag is often damaged and precious cell suspensions are lost.

[0006]

SUMMARY OF THE INVENTION The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a method and an apparatus for preventing a double-room freezing bag from being damaged when the bag is thawed.

[0007]

Means for Solving the Problems The present inventors have made intensive studies to solve the above-mentioned problems, and as a result, especially when there is a large difference in the accommodation amount between a plurality of chambers, the cell suspension of a specified volume as a whole is considered. Even if liquid is contained, for some reason,
For example, when filling the cell suspension, if the position of the small room of the multi-compartment freezing bag is lower than the position of the large room, the balance of the storage capacity between the rooms will be disrupted, and the cell suspension exceeding the specified capacity will fill the small room. The present inventors have found that the communication channel may be sealed in the housed state, and that the excessively housed cell suspension may cause breakage of the bag during freezing, and arrived at the present invention. That is, the present invention is a method for sealing a communication channel of a multi-chamber freezing bag, wherein sealing is performed while regulating the volume of a cell suspension to be contained to a specified volume or less. Here, when the double-chamber freezing bag is composed of two large and small chambers, at least the capacity of the small chamber may be regulated to a specified capacity or less. Further, the present invention, a mounting table for horizontally mounting a multi-chamber freezing bag, and means for regulating the amount of cell suspension contained in the multi-chamber freezing bag mounted on the mounting table,
A sealing means for sealing a communication channel of the multi-compartment freezing bag. Here, as the sealing means, a heat sealer or a high-frequency welder is used corresponding to the material for forming the double-chamber frozen bag.

[0008]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Next, embodiments of the present invention will be described with reference to the drawings. FIG. 1 is a plan view showing an embodiment of a communication channel sealing device for a multi-chamber freezing bag of the present invention, and FIG. 2 is a plan view showing an example of a multi-chamber freezing bag applied to the sealing device shown in FIG. FIG. As shown in FIG. 1, a sealing device 1 of the present invention includes, as basic components for carrying out the sealing method of the present invention, a mounting table 11 on which a multi-chamber freezing bag 2 is horizontally mounted, A means (lid 12 and lock 13) for regulating the amount of the cell suspension contained in the multi-compartment freezing bag 2 mounted on the mounting table 11;
And a sealing means (impulse sealer 15) for sealing the communication channels 24 and 25.

The double-chamber freezing bag shown in FIG. 2 is a so-called three-dimensional freezing bag having a flat rectangular chamber, and has a large-capacity chamber 21, a small-capacity chamber 22, and a communication part 23. Communication channels 24 and 25 for communicating the two chambers 21 and 22 are formed at both ends of the. The large chamber 21 is provided with a cell suspension collecting means 26 and a cell suspension outlet 27, and the small chamber 22 is provided with a cell suspension outlet 28. The mounting table 11 is provided with a chamber for accommodating the double-compartment freezing bag 2, and the chamber is partitioned into two large and small flat rectangular chambers 111 and 112 by a heater table 151 of the interposed impulse sealer 15. Omuro 111
The small chamber 112 has a similar shape and the same size to the large chamber 21 and the small chamber 22 of the multi-compartment freezing bag 2, respectively.
The communication part 23 is determined to be in close contact when the communication part 23 is accommodated in the mounting table 11.

The mounting table 11 has a cell suspension (not shown) accommodated in the multi-chamber freezing bag 2 mounted thereon.
As a means for regulating the amount of
2 is provided with an openable and closable lid 12 for regulating the amount of the cell suspension, and a lock 13 for fixing the lid 12, and the lock 13 is a pin 14 for fixing this to the mounting table 11.
Can be turned around the axis. Incidentally, the combination of the lid and the lock means may be provided also on the large chamber 111 side. As the cell suspension holding amount regulating means, in addition to the combination of the lid 12 and the lock 13, for example, the expansion of the small chamber at an appropriate position Various methods can be adopted, such as a pressing member (not shown) for preventing the pressure.

The chamber of the mounting table 11 has an impulse sealer 15
Is divided into a large room 111 and a small room 112 by the heater stand 151. The impulse sealer 15 is a heat sealer as a sealing means for sealing the communication channels 24 and 25 of the multi-compartment freezing bag 1, and an object to be welded is provided between the heater base 151 having the heater 153 and the heater base 151. It consists of a holding tool 152 for holding and holding
When the object to be welded is pressed against the heater base 151 by the holding tool 152, the heater 153 is heated. The heat sealer is preferably used when the material forming the bag is an olefin-based plastic, but a heating mold or the like can be used in addition to the impulse sealer. Also,
When the bag is made of a plastic having polarity such as vinyl chloride, nylon, or vinyl acetate,
A high frequency welder is preferably employed.

[Example 1] A chamber having a capacity of 20 ml and a capacity of 5 m
10 three-dimensional double-chamber freezing bags (bag thickness 8 mm) as shown in FIG. 2 having a total volume of 25 ml each having 1 chamber were filled, and each bag was filled with 25 ml of water. After sealing the communication channel using a dedicated sealing device as shown below, place it in a stainless steel case (a flat container to keep the thickness of the bag constant), And immersed in liquid nitrogen and frozen. After freezing, remove the bag from liquid nitrogen and remove
The bag was immersed in a 7 ° C. warm bath and thawed. The bag was checked for breakage and water leakage. As a result, no breakage and water leakage were observed for all bags.

[Comparative Example 1] Using the same three-dimensional multi-compartment freezing bags as in Example 1 and filling each with 25 ml of water, the same freezing and thawing as in Example 1 without sealing the communication channel. The bag was checked for breakage and water leakage, and all bags were found to have been damaged and leaked water, despite the fact that the thickness of the liquid was kept constant in a stainless steel case. Was done.

[0014]

As is apparent from the above description, by employing the present invention, it is possible to prevent the double-chamber freezing bag from being damaged by freezing.

[Brief description of the drawings]

FIG. 1 is a plan view showing an embodiment of a communication channel sealing device for a multi-chamber freezing bag of the present invention.

FIG. 2 is a plan view showing an example of a multi-chamber freezing bag applied to the sealing device shown in FIG.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Sealing device 11 Mounting table 111 Large room 112 Small room 12 Lid 13 Lock 14 Pin 15 Impulse sealer (sealing means) 151 Heater table 152 Clamping tool 153 Heater 2 Multi-room freezing bag 21 Large room 22 Small room 23 Communication part 24, 25 Communication channel 26 Cell suspension collection means 27, 28 Cell suspension outlet

Claims (5)

[Claims] 1. A method for sealing a communication channel of a multi-chamber freezing bag, wherein sealing is performed while regulating the volume of a cell suspension to be contained to a specified volume or less. 2. The sealing method according to claim 1, wherein the double-chamber freezing bag comprises two chambers, large and small, and at least the capacity of the small chamber is regulated to a specified capacity or less. 3. A mounting table for horizontally mounting a multi-chamber freezing bag, means for regulating the amount of cell suspension contained in the multi-chamber freezing bag mounted on the mounting table, and a multi-chamber freezing bag. And a sealing means for sealing the communication channel of the multi-chamber freezing bag. 4. The sealing device according to claim 3, wherein the sealing means is a heat sealer. 5. The sealing device according to claim 3, wherein the sealing means is a high-frequency welder.