JP2019154758A - Thawing device and thawing method - Google Patents

Abstract

To provide a thawing device and a thawing method capable of quickly thawing a frozen matter of biological-origin contained in a container.SOLUTION: A thawing device 10 comprises: a liquid flow path 16 connectable to a container 12; and a storage part 18 capable of storing a liquid L for thawing a frozen matter of biological origin 14. The device supplies the liquid L from the storage part 18 to the container 12 through the liquid flow path 16 and injects the liquid L into the container 12, thereby accelerating thawing of the frozen matter of biological origin 14.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a thawing apparatus and a thawing method for thawing a frozen substance derived from a living body stored in a container.

  Conventionally, a method of thawing (thawing) a frozen substance derived from a living body such as a frozen cell housed in a container such as a bag is performed by immersing the container in a constant temperature water bath. However, such a melting method using warm water is concerned about hygienic problems. Therefore, Patent Document 1 proposes an apparatus for sandwiching a container from above and below by two heating bags and thawing a frozen substance derived from a living body in the container.

European Patent No. 0318924

  However, in the prior art, heat transfer from the heating bag to the container is not efficiently performed, and it is considered that it takes a long time to thaw the biologically derived frozen material.

  The present invention has been made in consideration of such problems, and provides a melting apparatus and a melting method capable of hygienically and rapidly melting frozen living organisms contained in a container. Objective.

  In order to achieve the above object, one aspect of the present invention is a melting apparatus for thawing a biological frozen substance housed in a container, the liquid channel part connectable to the container, and the biological frozen A storage section capable of storing a liquid for melting an object, and supplying the liquid from the storage section to the container via the liquid flow path section and injecting the liquid into the container Is.

  According to this thawing device, since the thawing liquid is injected into the container in which the biologically-derived frozen material is accommodated, thawing of the biologically-derived frozen material is promoted. Therefore, it becomes possible to quickly thaw the frozen substance derived from the living body stored in the container. Moreover, since the heating liquid does not directly touch the outer surface of the container, it is hygienic.

  The melting apparatus may include a heating unit that heats the liquid to be injected into the container.

  With this configuration, the living body-derived frozen material is heated by the liquid, so that the living body-derived frozen material can be thawed more rapidly.

  In the melting apparatus, the liquid flow path section includes a supply flow path member that feeds the liquid from the storage section to the container, and a return reflux path member that returns the liquid from the container to the storage section. You may have.

  With this configuration, the liquid circulates in a circuit including the storage unit, the supply flow path member, the container, and the return reflux path member, so that the biologically derived frozen material can be more efficiently melted.

  In the above thawing apparatus, the biologically-derived frozen material is a frozen cell suspension containing a preservation solution, and the liquid may be a liquid that is inactive with respect to biologically-derived cells.

  With this configuration, it is possible to dilute the preservation solution at the same time as accelerating the thawing of the biologically derived frozen material, so that the work after thawing can be made more efficient.

  Another aspect of the present invention is a thawing method for thawing a frozen substance derived from a living body accommodated in a container, the step of preparing the container containing the frozen substance derived from the living body, and the frozen substance derived from the living body. It includes a step of preparing a liquid for melting, and an injection step of injecting the liquid into a container.

  The melting method may include a heating step of heating the liquid to be injected into the container.

  In the melting method, the liquid may be supplied from the storage unit to the container, and the liquid may be returned from the container to the storage unit.

  In the thawing method, the biological frozen substance is a frozen cell suspension containing a preservation solution, and in the injection step, an inactive liquid is injected into the biological cell as the liquid. May be.

  In the melting method, the injection step may be performed after a part of the living body-derived frozen material is melted in the container.

  According to the thawing device and the thawing method of the present invention, it is possible to hygienically and quickly thaw a living body-derived frozen material stored in a container.

1 is a schematic view of a melting apparatus according to a first embodiment of the present invention. It is the schematic of the melting apparatus which concerns on 2nd Embodiment of this invention. It is the schematic of the melting apparatus which concerns on 3rd Embodiment of this invention.

  Hereinafter, preferred embodiments of the melting apparatus and the melting method according to the present invention will be described with reference to the accompanying drawings.

  A thawing apparatus 10 according to the first embodiment shown in FIG. 1 is used for thawing a living body-derived frozen material 14 accommodated in a container 12.

  The biologically-derived frozen material 14 accommodated in the container 12 is obtained by freezing a liquid containing a biologically-derived material (biologically-derived liquid material). Examples of biological liquids include cell suspensions, blood, and plasma. Examples of the cell suspension include hematopoietic stem cells (umbilical cord blood, bone marrow fluid, peripheral blood stem cells, etc.) used for stem cell transplantation. The cells of the cell suspension are not limited to this, but myoblasts, cardiomyocytes, fibroblasts, synovial cells, epithelial cells, endothelial cells, hepatocytes, pancreatic cells, kidney cells, adrenal cells, Periodontal ligament cells, gingival cells, periosteum cells, skin cells, chondrocytes and other adherent cells, whole blood, red blood cells, white blood cells, lymphocytes (T lymphocytes, B lymphocytes), dendritic cells, plasma, platelets and platelets Blood cells such as plasma and blood components, bone skeleton mononuclear cells, hematopoietic stem cells, ES cells, pluripotent stem cells, iPS cell-derived cells (for example, iPS cell-derived cardiomyocytes), mesenchymal stem cells (for example, bone marrow, Adipose tissue, peripheral blood, skin, hair root, muscle tissue, endometrium, placenta, umbilical cord blood, etc.), sperm cells and egg cells. These cells may be cells into which genes used for gene therapy or the like have been introduced.

  In the first embodiment, the biologically-derived frozen material 14 is a frozen cell suspension containing a preservation solution (cryoprotectant). Examples of the preservation solution include DMSO, glycerin, trehalose, serum and the like, but are not limited thereto.

  The container 12 is, for example, a bag-like flat soft bag formed of a transparent resin film. The container 12 has a flat shape as a whole, and is formed in a square shape in plan view. The container 12 may be formed in a shape other than a quadrangular shape in plan view, for example, a circular shape or an elliptical shape. The container 12 may have a shape other than the flat shape. The container 12 may be made of a hard material.

  Specifically, the container 12 includes a bag-shaped storage chamber forming portion 12a that forms a storage chamber 12r therein, and a plate-shaped peripheral portion 12b that surrounds the outer periphery of the storage chamber forming portion 12a. A first port 13a and a second port 13b communicating with the storage chamber 12r are provided in the peripheral edge portion 12b.

  The thawing device 10 includes a liquid flow path section 16 that can be connected to the container 12 and a storage section 18 that can store the liquid L for thawing the living body-derived frozen material 14. The liquid L is fed to the container 12 via the and the liquid L is injected into the container 12.

  The liquid flow path unit 16 returns the liquid L from the container 12 to the storage unit 18 and the supply tubes 20 and 21 as supply flow path members that supply the liquid L from the storage unit 18 to the container 12. And a return tube 22 as a road member. The feed flow path member and the return flow path member are not limited to the form of a tube, but may be any members provided with a flow path.

  A first connector 23 that can be attached to and detached from the first port 13 a of the container 12 is provided at the first end 20 a that is one end of the feeding tube 20. The second end 20 b, which is the other end of the feeding tube 20, is connected to the liquid feeding pump 28. The liquid feed pump 28 is connected to a first end 21 a which is one end of another feed tube 21. The second end 21b, which is the other end of the feed tube 21, is connected to the storage 18 in the form of a tank.

  The liquid feed pump 28 is preferably one that can set the flow rate. In the first embodiment, the liquid feed pump 28 is a feed flow path member that feeds the liquid L from the reservoir 18 to the container 12 in order to form a liquid feed line between the reservoir 18 and the container 12. It can also be regarded as a part.

  When the liquid feeding pump 28 is operated, the liquid L in the storage unit 18 is sucked through the feeding tube 21. The sucked liquid L flows into the liquid feeding pump 28 and is then delivered to the feeding tube 20. The liquid L is supplied into the container 12 through the supply tube 20. The temperature of the liquid L is higher than at least the temperature of the biological frozen substance 14 in the container 12 before being injected into the container 12.

  In the case of the first embodiment in which the frozen cell suspension is the living body-derived frozen material 14, the liquid L is inactive with respect to the living body-derived cells, that is, does not adversely affect the living body-derived cells. For example, liquid medium, physiological saline, isotonic solution, buffer solution, Ringer's solution, Hank's balanced salt solution and the like can be mentioned.

  Here, the liquid medium refers to a basal medium and a basal medium to which biological serum is added. Examples of the basal medium include, but are not limited to, DMEM, MEM, F12, DME, RPMI 1640, MCDB (MCDB102, 104, 107, 131, 153, 199, etc.), L15, SkBM, RITC80-7, and the like. It is not a thing.

  A first connector 22, which is one end of the return tube 22, is provided with a second connector 24 that can be attached to and detached from the second port 13 b of the container 12. A second end 22 b that is the other end of the return tube 22 is connected to the storage unit 18. The liquid L that has flowed out of the container 12 is returned to the storage unit 18 via the return tube 22.

  In the case of the first embodiment, the storage unit 18 is provided with a heating unit 30 for heating the liquid L to be injected into the container 12. The illustrated heating unit 30 is configured to directly heat the liquid L stored in the storage unit 18. The heating unit 30 may be provided outside the storage unit 18 and configured to heat the liquid L in the storage unit 18 via the wall of the storage unit 18. The warming unit 30 warms the liquid L in the storage unit 18 to a predetermined temperature (for example, 30 to 40 ° C.).

  The heating unit 30 may be provided at a place other than the storage unit 18. For example, the liquid L may be heated on the path of the liquid L (the supply tube 20 or the supply tube 21) between the storage unit 18 and the container 12. The heating unit 30 is not essential and may not be provided. That is, the liquid L may not be positively heated.

  Next, the operation of the melting apparatus 10 according to the first embodiment configured as described above will be described.

  As shown in FIG. 1, the supply tube 20 and the return tube 22 are connected to the first port 13a and the second port 13b of the container 12 in which the living body-derived frozen material 14 is accommodated, respectively. Thereby, the circuit (circulation circuit) which consists of the storage part 18, the feed flow path member (feed tubes 20, 21 and the liquid feed pump 28), the container 12, and the return reflux path member (return tube 22) is comprised.

  Next, the container 12 in which the biologically-derived frozen material 14 is stored is left in a temperature environment higher than that of the biologically-derived frozen material 14, and the biologically-derived frozen material in the container 12 is filled to such an extent that the liquid L can be injected into the container 12. Wait for 14 to melt (initial melting step). The thawed state of the living body-derived frozen material 14 in the container 12 can be determined, for example, by visually confirming the state in the container 12. In this case, as indicated by a virtual line in FIG. 1, a heating mechanism 32 such as a heater is separately provided, and a process of heating the living body-derived frozen material 14 in the container 12 by the heating mechanism 32 is performed. The initial stage melting of the object 14 may be promoted.

  Next, when the liquid feeding pump 28 is operated, the liquid L heated by the warming unit 30 in the storage unit 18 is fed to the container 12 through the feeding tube 20 and into the container 12. Injected (injection process). The liquid L contacts the frozen living body 14 in the container 12 to heat the frozen living body 14. The heated living body-derived frozen material 14 is melted to become a living body-derived liquid material, and is diluted by mixing with the liquid L. The liquid L (the liquid derived from the living body diluted with the liquid L) flows out of the container 12 and returns to the storage unit 18 via the return tube 22. The liquid L that has returned to the storage unit 18 is heated again by the heating unit 30, and is sucked from the supply tube 21 again.

  As described above, the injection of the liquid L into the container 12 is continued for a certain period of time, so that the entire living body-derived frozen material 14 in the container 12 is melted.

  The melting device 10 according to the first embodiment has the following effects.

  According to the thawing device 10, since the thawing liquid L is injected into the container 12 in which the living body-derived frozen material 14 is accommodated, the melting of the living body-derived frozen material 14 is promoted. Therefore, the living body-derived frozen material 14 accommodated in the container 12 can be quickly thawed. Further, unlike a melting apparatus using a conventional constant temperature water tank, the heating liquid (warm water or the like) does not directly touch the outer surface of the container 12, so that it is hygienic.

  The melting apparatus 10 includes a heating unit 30 that heats the liquid L to be injected into the container 12. With this configuration, the living body-derived frozen material 14 is effectively heated by the liquid L, so that the living body-derived frozen material 14 can be thawed more rapidly.

  The liquid flow path section 16 includes a supply flow path member that supplies the liquid L from the storage section 18 to the container 12, and a return reflux path member that returns the liquid L from the container 12 to the storage section 18. With this configuration, since the liquid L circulates in a circuit including the storage unit 18, the supply flow path member, the container 12, and the return reflux path member, the living body-derived frozen substance 14 can be more efficiently melted.

  The biologically derived frozen material 14 is a frozen cell suspension containing a preservation solution, and the liquid L is a liquid L that is inactive with respect to biologically derived cells. Thereby, since the preservation | save liquid can be diluted simultaneously with accelerating | melting of the biologically derived frozen material 14, the efficiency of the operation | work after thawing | decompression is achieved.

  The melting device 40 according to the second embodiment shown in FIG. 2 is configured to store the liquid L flowing out of the container 12 in another container instead of returning it to the storage unit 18.

  Specifically, the melting device 40 includes a discharge tube 42 that discharges the liquid L from the second port 13 b of the container 12, and a recovery container 44 that stores the liquid L discharged from the discharge tube 42. A second connector 24 that can be attached to and detached from the second port 13 b is provided at a first end 42 a that is one end of the discharge tube 42. A second end 42 b that is the other end of the discharge tube 42 is connected to the collection container 44.

  Also by the melting device 40 according to the second embodiment, the melting of the living body-derived frozen material 14 is promoted by the injection of the liquid L into the container 12. Therefore, similarly to the melting device 10 according to the first embodiment, the living body-derived frozen material 14 accommodated in the container 12 can be melted hygienically and quickly.

  The melting device 50 according to the third embodiment shown in FIG. 3 has a common flow path for supplying the liquid L from the storage unit 18 to the container 12 and a flow path for returning the liquid L from the container 12 to the storage unit 18. It has become.

  Specifically, the melting device 50 includes a liquid flow path portion 52 that can be connected to the container 12 and a storage portion 18 that can store the liquid L. The liquid flow path section 52 includes a pump 54 capable of changing the liquid L feeding direction, a tube 58 having one end connected to the first port 13a of the container 12 via the connector 56 and the other end connected to the pump 54. A tube 59 having one end connected to the pump 54 and the other end connected to the reservoir 18. The second port 13b of the container 12 may be omitted.

  When the living body-derived frozen material 14 in the container 12 is thawed using the melting device 50, the tube 58 is connected to the first port 13a of the container 12 as shown in FIG. Next, it waits for the living body-derived frozen material 14 in the container 12 to melt to such an extent that the liquid L can be injected into the container 12. When the initial thawing of the living body-derived frozen material 14 in the container 12 is confirmed by the same method as in the first embodiment, the pump 54 is operated, and the liquid L is supplied from the reservoir 18 through the tubes 59 and 58 in order. The liquid L is injected into the container 12. Part of the living body-derived frozen material 14 is melted by the injected liquid L.

  After the liquid L is injected into the container 12 to some extent, next, the liquid feeding direction of the pump 54 is changed to the reverse direction. As a result, the liquid L (biologically derived liquid diluted with the liquid L) is sucked from the container 12 and returned from the container 12 to the storage unit 18 through the tubes 58 and 59 in order. By alternately repeating the injection operation and the return operation of the liquid L, the entire living body-derived frozen material 14 in the container 12 is melted.

  Also by the melting device 50 according to the third embodiment, the melting of the living body-derived frozen material 14 is promoted by the injection of the liquid L into the container 12. Therefore, similarly to the melting device 10 according to the first embodiment, the living body-derived frozen material 14 accommodated in the container 12 can be melted hygienically and quickly.

  The present invention is not limited to the above-described embodiments, and various modifications can be made without departing from the gist of the present invention.

DESCRIPTION OF SYMBOLS 10, 40, 50 ... Melting | fusing apparatus 12 ... Container 14 ... Living body origin frozen material 16, 52 ... Liquid flow path part 18 ... Storage part 20, 21 ... Feeding tube 22 ... Return tube 28 ... Liquid feeding pump 30 ... Heating part L ... Liquid

Claims (9)

A thawing device for thawing a frozen substance derived from a living body contained in a container,
A liquid channel portion connectable to the container;
A reservoir capable of storing a liquid for thawing the biologically derived frozen material,
A melting apparatus that feeds the liquid from the reservoir to the container via the liquid channel and injects the liquid into the container. The melting apparatus according to claim 1, wherein
A melting apparatus comprising a heating unit for heating the liquid to be injected into the container. The melting apparatus according to claim 1 or 2,
The liquid channel section includes a feeding channel member that feeds the liquid from the storage section to the container, and a return reflux path member that returns the liquid from the container to the storage section. . In the melting device according to any one of claims 1 to 3,
The biologically frozen product is a frozen cell suspension containing a preservation solution,
The melting device, wherein the liquid is a liquid that is inactive with respect to a living cell. A thawing method for thawing a frozen substance derived from a living body contained in a container,
Preparing the container in which the living body-derived frozen material is stored;
Preparing a liquid for thawing the biologically-derived frozen material;
An injection step of injecting the liquid into the container;
A melting method. The melting method according to claim 5, wherein
A melting method comprising a heating step of heating the liquid to be injected into the container. The melting method according to claim 5 or 6,
A melting method in which the liquid is supplied from the storage part to the container, and the liquid is returned from the container to the storage part. In the melting method according to any one of claims 5 to 7,
The biologically frozen product is a frozen cell suspension containing a preservation solution,
In the injecting step, a melting method in which an inactive liquid is injected into the living body-derived cell as the liquid. In the melting method according to any one of claims 5 to 8,
A thawing method, wherein the injection step is performed after a part of the living body-derived frozen material is melted in the container.

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