WO2006059626A1 - 生物学的試料保存用デバイス - Google Patents
生物学的試料保存用デバイス Download PDFInfo
- Publication number
- WO2006059626A1 WO2006059626A1 PCT/JP2005/021962 JP2005021962W WO2006059626A1 WO 2006059626 A1 WO2006059626 A1 WO 2006059626A1 JP 2005021962 W JP2005021962 W JP 2005021962W WO 2006059626 A1 WO2006059626 A1 WO 2006059626A1
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- WO
- WIPO (PCT)
- Prior art keywords
- biological sample
- sample
- cryopreservation container
- storage device
- biological
- Prior art date
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0236—Mechanical aspects
- A01N1/0263—Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving, e.g. cool boxes, blood bags or "straws" for cryopreservation
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0236—Mechanical aspects
- A01N1/0263—Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving, e.g. cool boxes, blood bags or "straws" for cryopreservation
- A01N1/0268—Carriers for immersion in cryogenic fluid, both for slow-freezing and vitrification, e.g. open or closed "straws" for embryos, oocytes or semen
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
- B01L3/5082—Test tubes per se
- B01L3/50825—Closing or opening means, corks, bungs
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/04—Closures and closing means
- B01L2300/046—Function or devices integrated in the closure
- B01L2300/047—Additional chamber, reservoir
Definitions
- the present invention is a device for storing a biological sample in a cryopreservation container, the sample mounting unit for placing the biological sample, and attached to the sample mounting unit
- the present invention relates to a biological sample storage device having a handle and having a structural force that can be stored in a cryopreservation container in at least a sample mounting portion of the device.
- An example of a rare specimen is an embryo egg.
- the importance of cryopreservation of embryos and eggs is pointed out as a key technology for infertility treatment programs.
- Vitality Collected stem cells and rare cells such as eggs are suspended in a cryopreservation solution to prepare a cell suspension, which is then stored and stored in a cell storage container. It was.
- An example of a collecting tool for collecting a trace amount of bodily fluid is the device disclosed in Patent Document 1.
- Patent Document 2 A cylindrical member holding tool for cryopreserving egg cells is disclosed in Patent Document 2.
- the work of attaching the vitrification solution of egg cells to this instrument is never allowed. It ’s not easy.
- the above work is performed using a microscope, it is necessary to carry out the work while holding the grip portion, and the work must be done with one hand.
- An example of a specimen that is difficult to handle is a membrane tissue.
- the membrane tissue include mucosal epithelial tissue, corneal epithelial tissue, cultured corneal tissue, and the like, all of which have a very thin film thickness.
- a storage Z transport method and a container therefor have been disclosed (Patent Documents 3 and 4) while maintaining the original shape of such a flat membrane structure without impairing biological characteristics.
- the method of Patent Document 3 is in contact with the wall surface of the container, heat is directly transmitted from the wall surface of the container, and thus the membrane tissue is damaged by rapid freezing.
- the container of Patent Document 4 can be stored under refrigerated conditions, but in the case of cryopreservation, the membrane may be torn by the backing member.
- Patent Document 1 Japanese Patent Laid-Open No. 2001-289844
- Patent Document 2 JP 2002-315573 A
- Patent Document 3 Japanese Patent Laid-Open No. 2003-9845
- Patent Document 4 Japanese Patent Laid-Open No. 2003-70460
- cryopreserving biological samples including rare and difficult-to-handle samples
- samples can be stored frozen without loss or damage, and frozen biological samples can be thawed easily. It is an object of the present invention to provide a biological sample storage device that can be used.
- a device for storing a biological sample in a cryopreservation container wherein the device includes a sample placement unit for placing a biological sample, and an attachment attached to the sample placement unit.
- a biological sample storage device having a hand and a structural force that at least a sample mounting portion of the device can be stored in the cryopreservation container.
- cryopreservation container is a vial
- handle portion of the device includes a cap structure for sealing the vial
- the material for the sample mounting part includes any one selected from polyethylene, polyester, polypropylene, polycarbonate, polystyrene, polymethyl methacrylate, polyimide, and polyamide. Device.
- a biological sample storage set comprising a cryopreservation container and a biological sample storage device comprising:
- the device comprises a sample placement part for placing a biological sample, and a handle part, The biological sample storage set, wherein at least the sample mounting portion of the device has a structural force that can be stored in the cryopreservation container.
- cryopreservation container is a vial
- handle portion of the device includes a cap structure for sealing the vial
- a method of storing a biological sample in a cryopreservation container using a device comprising a sample mounting part for placing a biological sample and a handle attached to the sample mounting part; J
- a method of storing a biological sample comprising:
- Specimen force contained in a biological sample 15.
- the biological sample storage device of the present invention When a biological sample is frozen using the biological sample storage device of the present invention (hereinafter sometimes referred to simply as "device"), a rare sample contained in the biological sample or a handling method is rarely included. It is possible to freeze difficult specimens without damaging them. The frozen biological sample can be easily thawed and collected without losing the contained specimen.
- the device of the present invention can facilitate work under a microscope.
- FIG. 1 Biological sample storage device with a recess in the sample mounting section (Example 1)
- FIG. 2 Biological sample storage device with a concave part on the specimen mounting part and a tapered part at the tip (Example 2)
- FIG. 3 Biological sample storage device including the biological sample storage device of Example 1 and a cryopreservation container (Example 3)
- FIG. 4 Biological sample storage set including the biological sample storage device of Example 1 and a cryopreservation container, wherein the living body in which the handle of the device and the cap of the cryopreservation container are integrated. Sample storage set (Example 4)
- the "biological sample storage device” has a distal end side and a base side, and includes a sample mounting portion for placing the biological sample on the distal end side.
- a handle portion attached to the sample mounting portion is provided.
- the sample mounting part can be stored in a cryopreservation container, and a biological sample is placed on the sample mounting part, and the cryopreservation container
- the biological sample can be stored frozen in a stored state. If the shape of the sample mounting portion of the device of the present invention can be stored in a container, it is acceptable.
- the sample placement part is stored in the cryopreservation container
- the specimen placement part in which the biological sample is placed is inserted and placed in the cryopreservation container body, and the cryopreservation container body is sealed. Sealing with possible caps.
- “At least the sample placement part is stored in the cryopreservation container.
- the term “payment” may include a cap structure of a cryopreservation container by having a structure in which the handle portion provided in the device of the present invention is fitted or screwed to the cryopreservation container main body as will be described later.
- the sample mounting part of the device of the present invention is stored in the cryopreservation container, and the handle part functions as a cap for sealing the cryopreservation container and may not be stored in the cryopreservation container. It is possible.
- “Placing a biological sample” means a state in which the biological sample is in contact with the sample mounting portion of the device, and preferably the biological sample is not dropped from the sample mounting portion. This means a state. For example, it includes dropping a sample liquid or placing a sample containing a tissue to attach the tissue.
- the “biological sample” refers to a sample containing a biological specimen such as a cell collected from a living body, a cultured cell, a tissue derived from a living body, or a cultured tissue.
- a biological specimen such as a cell collected from a living body, a cultured cell, a tissue derived from a living body, or a cultured tissue.
- the term “specimen” refers to the above-described biological specimen, which is a rare specimen or a minute specimen that is difficult to handle or a specimen that is difficult to handle due to the nature of the specimen.
- the specimen is a cell
- specific examples include embryos, egg cells, sperm cells, stem cells, and the like.
- the origin of such cells is not particularly limited, but human-derived cells are preferred.
- a membrane tissue when the specimen is a tissue, a membrane tissue is exemplified, and examples thereof include a mucosal epithelial tissue, a corneal epithelial tissue, and a cultured corneal tissue. Each of these has a very thin film thickness and is difficult to handle even if the film itself is to be used effectively.
- An example of the membrane tissue of the present invention is the cornea, which can also be applied to cryopreservation of the cornea for corneal transplantation.
- the "cryopreservation container” is not particularly limited as long as it is a container usually used for cryopreservation of the above-described biological sample, for example, cells.
- Vials or bags can be used.
- An example of a vial is a commercially available polypropylene cryovial capable of storing cells and the like.
- the bag include a cryopreservation bag comprising a laminate film of an ultrahigh molecular weight polyethylene film disclosed in JP-A-08-173505 and the like and a resin film compatible with the ultrahigh molecular weight polyethylene. be able to.
- sample placement part refers to a part for placing the biological sample.
- the shape of the sample mounting portion can be a film shape or a plate shape.
- the sample mounting part Preferably, the material or structure prevents the biological sample from dropping off on the sample mounting portion.
- the material for the sample mounting part include selecting a material having a pure water contact angle of about 60 to 90 degrees, and providing the concave part as the shape of the sample mounting part.
- the depth of the recess is preferably about 4 mm to 4 mm considering the size that the biological sample does not fall out and the device can be stored in the cryopreservation container.
- a tapered portion at the tip of the sample mounting portion.
- the tapered portion for example, when thawing an ovum cell, when immersing the device in mineral oil and rapidly thawing, the mineral oil having a large surface tension is likely to come into contact with cells existing on the device recess. Become.
- the material for the sample mounting portion of the present invention is preferably a material that can withstand cryopreservation. However, since the environment in the cryopreservation container is mild, it has a cold resistance like that of the cryopreservation container. Even a somewhat low material can be used. In addition, considering that a rare cell is placed on the material of the sample mounting portion, it is preferable that the material passes the safety test of the medical device, can be sterilized, and has cold resistance.
- the material satisfying these conditions can be selected from, for example, polyethylene, polyester, polypropylene, polycarbonate, polystyrene, polymethyl methacrylate, polyimide and polyamide. Among these, polyethylene terephthalate is preferable from the viewpoint of easy production, but the present invention is not limited thereto.
- the material of the sample mounting portion of the present invention is preferably an amorphous polymer from the viewpoint of enabling observation with a microscope.
- the amorphous polymer in the present invention refers to a material having a crystallinity of about 30% or less, preferably about 15% or less, particularly preferably about 5% or less, and visible light (380 to 780 nm) of a molded product.
- the transmittance of at least a part of the region is about 80% or more, preferably 90% or more, particularly preferably 95% or more. Those skilled in the art can appropriately control the crystallinity and transmittance according to molding conditions and the like.
- the "handle portion” refers to a portion where the device of the present invention can be handled without being in direct contact with the sample placement portion on which the biological sample is placed, for example, an index finger and a thumb. Or the thing which is a structure which enables pinching with tweezers etc. is said.
- the shape of the handle is smaller than that of the sample holder from the viewpoint of easy handling of the device. It is preferable.
- the present invention is not limited to these.
- the handle portion of the device of the present invention can further have a structure that fits or is screwed to the cryopreservation container body. By having a strong structure, it includes a cap structure for sealing the cryopreservation container, and the handle can be integrated with the cap of the cryopreservation container (see Fig. 4).
- sample mounting part and the handle part are integrally formed of the same material unless there are special circumstances.
- the device molding method of the present invention include extrusion molding, melt molding, injection molding, compression molding, and vacuum molding. Of these, injection molding and vacuum molding are preferred from the viewpoint of easy molding, but are not limited thereto.
- the device of the present invention can be provided as a “biological sample storage device set” by combining with a cryopreservation container.
- cryopreservation container when the cryopreservation container is a vial, a biological sample storage device and a cryopreservation container (a cryopreservation container body and a cap) can be used as a set.
- the handle portion of the device is provided with a structure that fits or is screwed to the cryopreservation container main body, and is integrated with the cap of the cryopreservation container, that is, includes a sample mounting portion and a cap structure.
- a biological sample storage device and a cryopreservation container main body integrally formed with a handle can be used as a set. As a result, the sample mounting portion of the device can be easily stored in the cryopreservation container.
- the present invention extends to a method for cryopreserving biological samples.
- the biological sample cryopreservation method uses at least the device of the present invention comprising a sample placement unit for placing a biological sample and a handle attached to the sample placement unit. According to the method including the following steps.
- the step of placing a biological sample on the sample mounting part The biological sample is brought into contact with the sample mounting portion of the existing device, and preferably, the processing step is performed so that the biological sample is not dropped from the sample mounting portion.
- the processing step is performed so that the biological sample is not dropped from the sample mounting portion.
- the specimen contained in the biological sample is, for example, a cell
- this step is achieved by dropping a solution in which the cell is suspended onto the sample mounting portion.
- the step can be achieved by placing the membrane tissue on a sample placement unit or winding the sample.
- the step of placing the biological sample and storing the sample mounting portion in the cryopreservation container means that, for example, when the cryopreservation container is a vial, the biological sample is placed.
- the sample holder is inserted and placed in the cryopreservation container body and sealed with a cap having a structure that can be fitted to the cryopreservation container body.
- it is important to insert it carefully so that the biological sample does not fall out of the sample holder.
- it can be stored in a cryopreservation container together with the sample placement part and handle.
- the preferred storage temperature is about -20 ° C or lower, preferably about -80 ° C. In the following, it is particularly preferably about ⁇ 196 ° C.
- the setting of these temperature conditions is achieved by, for example, a commercially available deep freezer, liquid nitrogen and dry ice. In particular, storage under liquid nitrogen that enables long-term storage, that is, storage at about -196 ° C is preferable.
- the cell when the specimen contained in the biological sample is a cell, the cell can be cryopreserved by a method including the following steps.
- the cells may be collected from vital force or collected from cultured cells.
- the number of cells in the cell suspension solution is not particularly limited. In the case of sperm cells, it is about 2 ⁇ 10 7 to 20 ⁇ 10 7 cells / ml, and in the case of egg cells, it is about 100 to 200 cells / ml. These concentration ranges may deviate from the range due to individual differences.
- the cryopreservation solution varies depending on the cell type and cannot be described uniformly, but a known cell culture medium or a cell culture medium developed in the future can be used. For example, a medium supplemented with about 10% dimethyl sulfoxide (DMSO) and 5 to 10% albumin as protective components is exemplified.
- DMSO dimethyl sulfoxide
- the cells can be cryopreserved using a known method, for example, a program freezing apparatus capable of controlling the temperature with liquid nitrogen.
- a program freezing apparatus capable of controlling the temperature with liquid nitrogen.
- the membrane tissue can be cryopreserved by a method including the following steps.
- the cornea can be obtained by cutting out from the eyeball by incision and removing unnecessary tissue.
- Step of infiltrating the cornea into a cryopreservation solution to prepare a biological sample The solvent used to wet the cornea mainly includes physiological saline, but is not limited thereto.
- the cornea can be cryopreserved using a known method, for example, a program freezing apparatus capable of controlling the temperature with liquid nitrogen.
- a program freezing apparatus capable of controlling the temperature with liquid nitrogen.
- the biological sample storage method of the present invention includes a method of thawing a cryopreserved sample. Specifically, thawing can be performed by immersing the device with the cryopreserved sample in Spitz containing mineral oil or medium.
- FIG. 1 shows a device (1) of the present invention.
- the device of this example is provided with a sample mounting part (11) on the tip side and a handle part (12) on the base side, and the sample mounting part (11) and the handle part (12) are integrated.
- the sample mounting part (11) is provided with a recess (111).
- This device can produce polyethylene terephthalate film by injection molding or vacuum molding.
- FIG. 2 is a view showing another embodiment of the device (1) of the present invention. Similar to the device of Example 1, the device of this example is provided with a sample mounting part (11) on the tip side and a handle part (12) on the base side, and a tapered part at the tip of the sample mounting part (11). (112). Deba The chair material and device manufacturing method are the same as in Example 1.
- FIG. 3 shows a biological sample storage set according to the present invention.
- the biological sample storage set shown in FIG. 3 includes the cryovial made of polypropylene, which is the device (1) of Example 1 and the cryopreservation container (2).
- the cryopreservation container (2) is composed of a cryopreservation container body (21) and a cap (22).
- the method for preserving cells using the biological sample storage set of this example is as follows.
- FIG. 4 is a diagram showing another embodiment of the biological sample storage set of the present invention.
- the biological sample storage set shown in Fig. 4 includes a sample placement part (11) using a polypropylene plate produced by extrusion molding, and a handle part (12) attached to this and including a cap structure of a cell storage container. And a cell storage container body (21).
- the device handle (12) and the cell storage container cap (22) are in a single body.
- the method for preserving cells using the biological sample storage set of this example is as follows.
- the sample holder (11) of the device Places the biological sample (A) on the sample holder (11) of the device. Next, the part (11) which is the cap (22) of the cell storage container and the handle (12) of the device is picked, and the sample mounting part (11) is inserted into the cell storage container body (21). After sealing with the cap (22), the cryopreservation container (2) is immersed in liquid nitrogen and stored frozen at a cryogenic temperature (about 1196 ° C).
- Example 5 Thawing Example 1 of Frozen Biological Sample
- Fig. 5 shows a frozen biological sample using the biological sample storage device (1) of Example 1, taken out of the cryopreservation container (2), and the freeze-thawing medium in Spitz. It is a figure which shows the thawing
- the temperature of the freezing and thawing medium (31) in Spitz may be adjusted to about 20-30 ° C.
- Example 6 Thawing Example 2 of Frozen Biological Sample
- Fig. 6 shows that the biological sample frozen using the biological sample storage device (1) of Example 2 is removed from the cryopreservation container (2) and immersed in mineral oil (32). It is a figure which shows the decompression
- the temperature of the mineral oil (32) should be adjusted to about 10-40 ° C.
- a sample containing a rare sample or a sample that is difficult to handle can be frozen and stored, and the frozen sample can be easily thawed. Can be done. Since the device of the present invention can be stored in an existing cryopreservation container, it can be cryopreserved and thawed easily and inexpensively. Furthermore, when the device of the present invention is used, a specimen that is difficult to handle, for example, a specimen such as a cornea can be stored frozen without being damaged, and can be thawed more easily.
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Priority Applications (1)
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JP2006547966A JP4876922B2 (ja) | 2004-12-03 | 2005-11-30 | 生物学的試料保存用デバイス |
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JP2004350714 | 2004-12-03 | ||
JP2004-350714 | 2004-12-03 |
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WO2006059626A1 true WO2006059626A1 (ja) | 2006-06-08 |
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Cited By (12)
Publication number | Priority date | Publication date | Assignee | Title |
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JP2008212062A (ja) * | 2007-03-05 | 2008-09-18 | Niwa Masami | 細胞凍結保存方法及び細胞培養方法 |
JP2009148221A (ja) * | 2007-12-21 | 2009-07-09 | National Agriculture & Food Research Organization | 生殖細胞凍結保存容器及び生殖細胞凍結保存方法 |
WO2016031916A1 (ja) * | 2014-08-29 | 2016-03-03 | 株式会社北里バイオファルマ | 採取生体組織凍結保存用具および組織片凍結保存方法 |
WO2015173809A3 (en) * | 2014-05-13 | 2016-03-31 | Mfc Global Ltd | Cell tray |
WO2016111217A1 (ja) * | 2015-01-09 | 2016-07-14 | 学校法人明治大学 | 中空糸凍結保存用具及び細胞凍結保存方法 |
CN111108375A (zh) * | 2017-09-21 | 2020-05-05 | 浜松光子学株式会社 | 激光解吸电离法和质量分析方法 |
US10638748B2 (en) | 2015-12-22 | 2020-05-05 | Corning Incorporated | Break away/tear away cryopreservation vial and methods for manufacturing and using same |
JP2020120627A (ja) * | 2019-01-31 | 2020-08-13 | 三菱製紙株式会社 | 細胞又は組織のガラス化凍結保存用治具 |
JP2021016336A (ja) * | 2019-07-19 | 2021-02-15 | 国立大学法人秋田大学 | 哺乳動物初期胚の凍結保存方法 |
US11008157B2 (en) | 2013-08-16 | 2021-05-18 | Corning Incorporated | Vessels and methods for cryopreservation |
US11684064B2 (en) | 2015-11-16 | 2023-06-27 | Corning Incorporated | Cryogenic vial assemblies |
WO2024027532A1 (zh) * | 2022-08-01 | 2024-02-08 | 深圳拜尔洛克生物技术有限公司 | 一种卵子或胚胎的解冻方法 |
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