WO2013128561A1 - Cell manipulation implement - Google Patents

Cell manipulation implement Download PDF

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Publication number
WO2013128561A1
WO2013128561A1 PCT/JP2012/054900 JP2012054900W WO2013128561A1 WO 2013128561 A1 WO2013128561 A1 WO 2013128561A1 JP 2012054900 W JP2012054900 W JP 2012054900W WO 2013128561 A1 WO2013128561 A1 WO 2013128561A1
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Prior art keywords
flow path
cell handling
handling tool
main body
container
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PCT/JP2012/054900
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French (fr)
Japanese (ja)
Inventor
鈴木　保彦
彩 大谷
康弘 横山
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株式会社メニコン
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Priority to PCT/JP2012/054900 priority Critical patent/WO2013128561A1/en
Publication of WO2013128561A1 publication Critical patent/WO2013128561A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/12—Well or multiwell plates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus

Definitions

  • the present invention relates to a cell handling tool.
  • the present invention has been made in order to solve the above-mentioned problems, and its main purpose is to easily prevent the contamination of bacteria while improving the handleability by preventing problems such as accidental sample spillage. Is to be able to do that.
  • the cell handling tool of the present invention contains cells, and includes a container body having a flat bottom, and an elastic arm part extending outward from the outer peripheral surface of the container body.
  • the container body is housed in a cylindrical container having a bottom represented by a general-purpose petri dish, and a part of the arm part is brought into contact with the inner peripheral surface of the cylindrical container.
  • the importance of individual sample management is very high and is preferably used.
  • the fixing method is simple and the risk of contamination is low as compared with the case of using a tape or an adhesive sheet.
  • the cylindrical container is covered, it is possible to easily prevent the bacteria from being mixed into the housing main body. Furthermore, since the handleability can be improved without changing the size of the container main body (without making it larger than a certain size), it is possible to reduce raw material costs and disposal burden.
  • the arm portion is formed in an outwardly convex arc shape. According to such a structure, an arm part can be made to follow the internal peripheral surface of a cylindrical container. Therefore, it can fix more reliably.
  • the arm portion is provided away from the bottom surface of the housing main body.
  • the inner bottom surface of the cylindrical container may have a step along the outer periphery due to manufacturing reasons. However, according to such a configuration, the arm portion does not hit the step on the inner bottom surface. Can be fixed to.
  • a part of the arm portion is brought into contact with an inner peripheral surface of a cylindrical container having a bottom, and the housing main body is fixed to the cylindrical container.
  • the cell handling tool of the present invention is used as a sperm separation tool.
  • the container main body is discharged from the first flow path for flowing the semen sample, the first supply section for supplying the semen sample to the first flow path, and the first flow path.
  • a second storage part for storing the fluid to be discharged, and the middle part of the second flow path is a merged flow path with the first flow path.
  • the present invention is suitably applied to such an open cell handling tool.
  • the housing portion main body by providing an elastic arm portion on the outer peripheral surface of the housing portion main body, the housing portion main body can be fixed to a cylindrical container having a bottom such as a petri dish, and handling properties are greatly improved. It can be managed as an integrated object with a cylindrical container (storage container). In addition, if the cylindrical container is covered, it is possible to easily prevent the bacteria from being mixed into the housing main body.
  • FIG. 1 It is a schematic perspective view which shows one Example of the cell handling tool of this invention. It is the figure which looked at the use condition of the cell handling tool of FIG. 1 from the top. It is a longitudinal cross-sectional view of the use condition (state which covered the storage container) of the cell handling tool of FIG. (A)-(f) is the schematic which shows the shape of the arm part in another embodiment.
  • FIG. 1 to 3 are schematic views showing an embodiment of the cell handling device of the present invention
  • FIG. 1 is a perspective view
  • FIG. 2 is a view of the usage state from above
  • FIG. It is a longitudinal cross-sectional view of a state.
  • the cell handling tool 1 includes a storage unit main body 2 that stores cells, and arm portions 3, 3, 3, and 3 that extend outward from the outer peripheral surface of the storage unit main body 2, and separate sperm. Used as a tool.
  • the accommodating portion main body 2 is formed in a substantially rectangular flat plate shape, and includes a fine first flow path 21 for flowing a fluid in the main surface direction and a fine second flow path 22.
  • the housing main body 2 is formed by stacking and bonding a flat bottom plate 11 and an upper plate 12, and the first flow path 21 and the second flow path 22 are formed on the top surface of the bottom plate 11. It is formed by forming a groove in. While the first flow path 21 is linear, the second flow path 22 joins the first flow path from the right side with respect to the flow direction, and the midstream portion 22b is connected to the first flow path 21. And the downstream portion 22c is branched to the right side.
  • the channel diameter of each channel is typically 10 to 1000 ⁇ m, preferably 50 to 400 ⁇ m.
  • the housing part 2 is provided with a first supply part 31 for supplying fluid to the first flow path 21 and a second supply part 32 for supplying fluid to the second flow path 22.
  • a first supply part 31 for supplying fluid to the first flow path 21
  • a second supply part 32 for supplying fluid to the second flow path 22.
  • Each supply part 31 and 32 is provided in the part corresponding to the starting end part of each flow path 21 and 22 of upper board 12, circular through-hole 13 formed in the thickness direction, and the upper surface of upper board 12, It is comprised by the cylinder part 14 connected with the through-hole 13.
  • the starting ends of the flow paths 21 and 22 are provided in the vicinity of the center of the through holes 13 and 13.
  • the inner diameter of the cylindrical portion 14 is substantially the same as the diameter of the through hole 13, and the starting end portions of the flow paths 21 and 22 are opened upward.
  • the volume of each supply unit is typically 10 to 1000 ⁇ l.
  • the storage unit body 2 includes a first storage unit 41 that stores fluid discharged from the first flow path 21, and a second storage unit 42 that stores fluid discharged from the second flow path 22.
  • a first storage unit 41 that stores fluid discharged from the first flow path 21, and a second storage unit 42 that stores fluid discharged from the second flow path 22.
  • Each storage portion 41, 42 is provided on the upper surface of the upper plate 12 with a circular through hole 15 formed in the thickness direction at a portion corresponding to the end portion of each flow path 21, 22 of the upper plate 12. It is comprised by the cylinder part 16 connected with the through-hole 15.
  • the end of each flow path 21, 22 is provided near the center of the through holes 15, 15.
  • the inner diameter of the cylindrical portion 16 is substantially the same as the diameter of the through hole 15, and the end portions of the flow paths 21 and 22 are opened upward.
  • the volume of each reservoir is typically 10 to 1000 ⁇ l.
  • a semen sample containing sperm is supplied to the first flow path 21, a medium is supplied to the second flow path 22, and the semen sample and the medium are brought into contact in a laminar flow state in the merge flow path 23.
  • motile sperm highly motile sperm contained in the semen sample
  • the semen sample from which the motile sperm has been separated is discharged to the first reservoir (waste liquid reservoir) 41, and the medium containing the motile sperm is discharged to the second reservoir (motile sperm reservoir) 42.
  • Motile sperm and non-motile sperm are separated.
  • the separation efficiency can be improved by making the cross-sectional area of the medium flow larger than the cross-sectional area of the semen sample flow.
  • one medium flow path for circulating the medium is used, but a plurality of medium flow paths may be formed.
  • the four arm portions 3, 3, 3, and 3 are integrally formed on the side surface of the housing portion main body 2 so as to be elastically deformable. How much and in what direction the arm 3 is formed in which position in which position of the housing main body 2 is set as appropriate. In this embodiment, both ends of the opposing side surfaces of the housing main body 2 are arranged.
  • the arc-shaped arm portions 3, 3, 3, and 3 that protrude outwardly from the portion extend, and the arm portions 3 and 3 that extend from the same side surface form a single arc shape.
  • the arm portion 3 extends from the left and right side surfaces 17 and 18 of the upper plate 12 constituting the housing portion main body 2 in the main surface direction of the upper plate 12, and the arm portion 3 is separated from the bottom surface of the housing portion main body 2. Is formed.
  • the arm portion 3 is formed at a height of 1 mm or more and 15 mm or less from the bottom surface of the housing portion main body 2.
  • both ends of the arm portion 3 are joined to the housing portion main body 2 to form a substantially semicircular shape or a substantially V shape.
  • the arc-shaped arm portions 3 and 3 extending from the same side surface are formed such that their free ends face in opposite directions.
  • a bulging portion 3 a bulging outward is formed at the tip of the arc-shaped arm portion 3.
  • the arm portion may be a latch member biased outward by a spring or the like.
  • the material of the housing part main body 2 and the arm part 3 is not particularly limited, but a thermoplastic resin is preferable from the viewpoint of moldability.
  • the thermoplastic resin include polyethylene (PE) such as high density polyethylene (HDPE), medium density polyethylene (MDPE), and low density polyethylene (LDPE), polypropylene (PP), polyvinylidene chloride, and polyvinyl chloride (PVC).
  • PE polyethylene
  • HDPE high density polyethylene
  • MDPE medium density polyethylene
  • LDPE low density polyethylene
  • PP polypropylene
  • PP polyvinylidene chloride
  • PVC polyvinyl chloride
  • PTFE Polytetrafluoroethylene
  • PS polystyrene
  • PVAc polyvinyl acetate
  • Teflon registered trademark
  • ABS resin acrylonitrile butadiene styrene resin
  • AS resin acrylonitrile styrene resin
  • PMMA etc.
  • General purpose plastics such as (meth) acrylic resin, polyethylene terephthalate (PET); polyamide (PA), polyacetal (POM), polycarbonate (PC), modified polyphenylene ether (modified PPE), polybutylene terephthalate (PBT), glass fiber reinforced polyethylene terephthalate (GF-PET), engineering plastics such as cyclic polyolefin (COP); polyphenylene sulfide (PPS), polytetrafluoroethylene (PTFE), polysulfone (PSF), polyether Examples include super engineering plastics such as sulfone (PES), amorphous polyarylate (PAR), liquid crystal polymer (LCP), polyether ether ketone (PEEK), thermoplastic polyimide (PI), and polyamideimide (PAI).
  • PES polyethylene terephthalate
  • PA polyamide
  • POM polyacetal
  • PC polycarbonate
  • PC modified polyphenylene ether
  • PBT polybutylene terephthalate
  • FIGS. 2 and 3 are schematic views showing the usage state of the cell handling tool 1 of this embodiment.
  • the cell handling tool 1 is placed on a cylindrical container 101 having a bottom typified by a general-purpose petri dish, and is fixed by bringing a part of an arm 3 that can be elastically deformed into contact with the inner peripheral surface of the cylindrical container 101.
  • the arm portion 3 is formed away from the bottom surface of the housing portion main body 2, even if the step 101 a is provided along the outer periphery of the inner bottom surface of the cylindrical container 101 as in the illustrated example, the arm portion 3 is not It does not hit the step 101a. In such a fixed state, the cells are separated (selected), cultured, stored, and the like.
  • the lid 102 of the cylindrical container 101 If the lid 102 of the cylindrical container 101 is used, it is possible to prevent the bacteria from being mixed into the housing body 2. Moreover, the bad influence (for example, influence by a solvent evaporating) to the cell accommodated in the accommodating part main body 2 can be suppressed.
  • the case of using as a sperm separation tool has been described.
  • it can be applied as a cell culture tool by providing a medium in the container body.
  • the cell handling device of the present invention can be suitably used as a device for separating (selecting), culturing and storing cells.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
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  • Biochemistry (AREA)
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Abstract

　Provided is a cell manipulation implement capable of preventing microbial contamination while also improving manipulation properties and preventing problems such as the accidental spillover of a sample.　The cell manipulation implement (1) according to the present application is equipped with: an accommodation part (2) that accommodates cells and has a tabular bottom part; and an arm part (3) that is elastic and extends from the outer peripheral surface of the accommodation part (2) to the outside.

Description

細胞取扱用具Cell handling tools
　本発明は、細胞取扱用具に関する。 The present invention relates to a cell handling tool.
　細胞の分離（選別）、培養、保存等に際し、処置の完全性や安全性を確保する観点から、菌の混入を抑制することが必要とされる。細胞の分離、培養、保存等を行う細胞取扱用具の中には、開放系で菌の混入に対して十分な対策がとられていないものもある。そこで、専用の収納容器に収納して菌の混入を防止することが提案されている（例えば、特許文献１）。 In the separation (sorting), culture, storage, etc. of cells, it is necessary to suppress the contamination of bacteria from the viewpoint of ensuring the completeness and safety of the treatment. Some cell handling tools for separating, culturing, storing, etc. of cells do not have sufficient measures against bacterial contamination in an open system. Thus, it has been proposed to prevent contamination of bacteria by storing in a dedicated storage container (for example, Patent Document 1).
　しかし、細胞取扱用具の形状・サイズは様々であり、専用の収納容器を作製するのはコスト面に問題がある。一方で、細胞取扱用具自体の取り扱い性の確保が望まれている（特に、細胞取扱用具が開放系の場合）。 However, there are various shapes and sizes of the cell handling tools, and there is a problem in terms of cost to produce a dedicated storage container. On the other hand, it is desired to ensure the handleability of the cell handling tool itself (particularly when the cell handling tool is an open system).
特開２０１２－２４０２０号公報JP 2012-24020 A
　本発明は、上記課題を解決するためになされたものであり、その主たる目的は、誤ってサンプルがこぼれてしまう等の不具合を防止して取り扱い性を向上させながら、菌の混入を容易に防止することができるようにすることにある。 The present invention has been made in order to solve the above-mentioned problems, and its main purpose is to easily prevent the contamination of bacteria while improving the handleability by preventing problems such as accidental sample spillage. Is to be able to do that.
　本発明の細胞取扱用具は、細胞を収容し、底部が平板状の収容部本体と、この収容部本体の外周面から外方に延設され、弾性を有する腕部とを備える。このような構成によれば、例えば、汎用のシャーレに代表される底部を有する円筒状容器に収納し、円筒状容器の内周面に腕部の一部を当接させることにより、収容部本体を円筒状容器に固定することができる。したがって、取り扱い性が格段に向上し、円筒状容器（収納容器）との一体物として管理することができる。医療機関では、試料の個別管理の重要性が非常に高く、好適に用いられる。しかも、テープや粘着シート等を用いる場合に比べて、固定方法も簡便である上に、異物混入のリスクが低い。また、円筒状容器に蓋をすれば、収容部本体への菌の混入を容易に防止することができる。さらに、収容部本体のサイズを変更することなく（一定以上の大きさにすることなく）、取り扱い性を向上させることができるので、原材料費や廃棄の負担を軽減することができる。 The cell handling tool of the present invention contains cells, and includes a container body having a flat bottom, and an elastic arm part extending outward from the outer peripheral surface of the container body. According to such a configuration, for example, the container body is housed in a cylindrical container having a bottom represented by a general-purpose petri dish, and a part of the arm part is brought into contact with the inner peripheral surface of the cylindrical container. Can be fixed to the cylindrical container. Therefore, the handleability is remarkably improved and can be managed as an integrated object with the cylindrical container (storage container). In a medical institution, the importance of individual sample management is very high and is preferably used. In addition, the fixing method is simple and the risk of contamination is low as compared with the case of using a tape or an adhesive sheet. In addition, if the cylindrical container is covered, it is possible to easily prevent the bacteria from being mixed into the housing main body. Furthermore, since the handleability can be improved without changing the size of the container main body (without making it larger than a certain size), it is possible to reduce raw material costs and disposal burden.
　好ましい実施形態においては、上記腕部が外方に凸の円弧形状に形成されている。このような構成によれば、腕部を円筒状容器の内周面に沿わせることができる。したがって、より確実に固定することができる。 In a preferred embodiment, the arm portion is formed in an outwardly convex arc shape. According to such a structure, an arm part can be made to follow the internal peripheral surface of a cylindrical container. Therefore, it can fix more reliably.
　好ましい実施形態においては、上記腕部が、上記収容部本体の底面から離間して設けられている。上記円筒状容器の内側底面には、製造の関係からその外周に沿って段差が設けられている場合があるが、このような構成によれば、腕部が内側底面の段差に当たらず、確実に固定することができる。 In a preferred embodiment, the arm portion is provided away from the bottom surface of the housing main body. The inner bottom surface of the cylindrical container may have a step along the outer periphery due to manufacturing reasons. However, according to such a configuration, the arm portion does not hit the step on the inner bottom surface. Can be fixed to.
　好ましい実施形態においては、底部を有する円筒状容器の内周面に上記腕部の一部を当接させて、上記収容部本体を前記円筒状容器に固定させる。このような実施形態によれば、上述のとおり、取り扱い性が格段に向上し、円筒状容器に蓋をすれば、収容部本体への菌の混入を容易に防止することができる。 In a preferred embodiment, a part of the arm portion is brought into contact with an inner peripheral surface of a cylindrical container having a bottom, and the housing main body is fixed to the cylindrical container. According to such an embodiment, as described above, the handleability is remarkably improved, and if the cylindrical container is covered, it is possible to easily prevent the bacteria from being mixed into the housing main body.
　好ましい実施形態においては、本発明の細胞取扱用具は、***分離用具として用いられる。具体的には、上記収容部本体は、***サンプルを流通させる第１の流路と、この第１の流路に***サンプルを供給する第１の供給部と、前記第１の流路から排出される***サンプルを貯留する第１の貯留部と、媒体を流通させる第２の流路と、この第２の流路に媒体を供給する第２の供給部と、前記第２の流路から排出される流体を貯留する第２の貯留部とを備え、前記第２の流路の中流部が前記第１の流路との合流流路とされている。本発明は、このような開放系の細胞取扱用具に対し、好適に適用される。 In a preferred embodiment, the cell handling tool of the present invention is used as a sperm separation tool. Specifically, the container main body is discharged from the first flow path for flowing the semen sample, the first supply section for supplying the semen sample to the first flow path, and the first flow path. A first storage part for storing the semen sample to be performed, a second flow path for circulating the medium, a second supply part for supplying the medium to the second flow path, and the second flow path And a second storage part for storing the fluid to be discharged, and the middle part of the second flow path is a merged flow path with the first flow path. The present invention is suitably applied to such an open cell handling tool.
　本発明によれば、収容部本体の外周面に弾性を有する腕部を設けることで、収容部本体をシャーレ等の底部を有する円筒状容器に固定することができ、取り扱い性が格段に向上し、円筒状容器（収納容器）との一体物として管理することができる。また、円筒状容器に蓋をすれば、収容部本体への菌の混入を容易に防止することができる。 According to the present invention, by providing an elastic arm portion on the outer peripheral surface of the housing portion main body, the housing portion main body can be fixed to a cylindrical container having a bottom such as a petri dish, and handling properties are greatly improved. It can be managed as an integrated object with a cylindrical container (storage container). In addition, if the cylindrical container is covered, it is possible to easily prevent the bacteria from being mixed into the housing main body.
本発明の細胞取扱用具の一実施例を示す概略斜視図である。It is a schematic perspective view which shows one Example of the cell handling tool of this invention. 図１の細胞取扱用具の使用状態を上から見た図である。It is the figure which looked at the use condition of the cell handling tool of FIG. 1 from the top. 図１の細胞取扱用具の使用状態（収納容器の蓋をした状態）の縦断面図である。It is a longitudinal cross-sectional view of the use condition (state which covered the storage container) of the cell handling tool of FIG. （ａ）から（ｆ）は、別の実施形態における腕部の形状を示す概略図である。(A)-(f) is the schematic which shows the shape of the arm part in another embodiment.
　以下、本発明の具体的実施例を図面に基づいて詳細に説明する。 Hereinafter, specific embodiments of the present invention will be described in detail with reference to the drawings.
　図１から図３は、本発明の細胞取扱用具の一実施例を示す概略図であり、図１は斜視図であり、図２は使用状態を上から見た図であり、図３は使用状態の縦断面図である。 1 to 3 are schematic views showing an embodiment of the cell handling device of the present invention, FIG. 1 is a perspective view, FIG. 2 is a view of the usage state from above, and FIG. It is a longitudinal cross-sectional view of a state.
　本実施例の細胞取扱用具１は、細胞を収容する収容部本体２と、収容部本体２の外周面から外方に延設された腕部３，３，３，３とを備え、***分離用具として用いられる。 The cell handling tool 1 according to the present embodiment includes a storage unit main body 2 that stores cells, and arm portions 3, 3, 3, and 3 that extend outward from the outer peripheral surface of the storage unit main body 2, and separate sperm. Used as a tool.
　収容部本体２は、略矩形の平板状とされ、その主面方向に流体を流通させる微細な第１の流路２１と、微細な第２の流路２２とを備える。本実施例では、収容部本体２は、平板状の底板１１と上板１２とを重ねて接着させることにより形成され、第１の流路２１および第２の流路２２は、底板１１の上面に溝を形成することにより形成されている。第１の流路２１は直線状とされる一方、第２の流路２２は流通方向に対して右側方から第１の流路に合流し、その中流部２２ｂが第１の流路２１との合流流路２３とされ、下流部２２ｃが右側方に分岐されている。各流路の流路径は、代表的には、１０～１０００μｍであり、好ましくは５０～４００μｍである。 The accommodating portion main body 2 is formed in a substantially rectangular flat plate shape, and includes a fine first flow path 21 for flowing a fluid in the main surface direction and a fine second flow path 22. In the present embodiment, the housing main body 2 is formed by stacking and bonding a flat bottom plate 11 and an upper plate 12, and the first flow path 21 and the second flow path 22 are formed on the top surface of the bottom plate 11. It is formed by forming a groove in. While the first flow path 21 is linear, the second flow path 22 joins the first flow path from the right side with respect to the flow direction, and the midstream portion 22b is connected to the first flow path 21. And the downstream portion 22c is branched to the right side. The channel diameter of each channel is typically 10 to 1000 μm, preferably 50 to 400 μm.
　収容部本体２には、第１の流路２１に流体を供給する第１の供給部３１と、第２の流路２２に流体を供給する第２の供給部３２とが設けられている。第１の供給部３１および第２の供給部３２の形状は適宜に設定されるが、本実施例では、円筒状に形成されている。各供給部３１，３２は、上板１２の各流路２１，２２の始端部に対応する部位に、厚さ方向に形成された円形の貫通孔１３と、上板１２の上面に設けられ、貫通孔１３と連通する筒部１４とで構成されている。各流路２１，２２の始端は、貫通孔１３，１３の中心部付近に設けられている。筒部１４の内径は、貫通孔１３の直径と略同一であり、各流路２１，２２の始端部は上方に開放されている。各供給部の容積は、代表的には、１０～１０００μｌである。 The housing part 2 is provided with a first supply part 31 for supplying fluid to the first flow path 21 and a second supply part 32 for supplying fluid to the second flow path 22. Although the shape of the 1st supply part 31 and the 2nd supply part 32 is set suitably, in the present Example, it is formed in the cylindrical shape. Each supply part 31 and 32 is provided in the part corresponding to the starting end part of each flow path 21 and 22 of upper board 12, circular through-hole 13 formed in the thickness direction, and the upper surface of upper board 12, It is comprised by the cylinder part 14 connected with the through-hole 13. The starting ends of the flow paths 21 and 22 are provided in the vicinity of the center of the through holes 13 and 13. The inner diameter of the cylindrical portion 14 is substantially the same as the diameter of the through hole 13, and the starting end portions of the flow paths 21 and 22 are opened upward. The volume of each supply unit is typically 10 to 1000 μl.
　収容部本体２には、第１の流路２１から排出される流体を貯留する第１の貯留部４１と、第２の流路２２から排出される流体を貯留する第２の貯留部４２とが設けられている。第１の貯留部４１および第２の貯留部４２の形状は適宜に設定されるが、本実施例では、円筒状に形成されている。各貯留部４１，４２は、上板１２の各流路２１，２２の終端部に対応する部位に、厚さ方向に形成された円形の貫通孔１５と、上板１２の上面に設けられ、貫通孔１５と連通する筒部１６とで構成されている。各流路２１，２２の終端は、貫通孔１５，１５の中心部付近に設けられている。筒部１６の内径は、貫通孔１５の直径と略同一であり、各流路２１，２２の終端部は上方に開放されている。各貯留部の容積は、代表的には、１０～１０００μｌである。 The storage unit body 2 includes a first storage unit 41 that stores fluid discharged from the first flow path 21, and a second storage unit 42 that stores fluid discharged from the second flow path 22. Is provided. Although the shape of the 1st storage part 41 and the 2nd storage part 42 is set suitably, in the present Example, it is formed in the cylindrical shape. Each storage portion 41, 42 is provided on the upper surface of the upper plate 12 with a circular through hole 15 formed in the thickness direction at a portion corresponding to the end portion of each flow path 21, 22 of the upper plate 12. It is comprised by the cylinder part 16 connected with the through-hole 15. As shown in FIG. The end of each flow path 21, 22 is provided near the center of the through holes 15, 15. The inner diameter of the cylindrical portion 16 is substantially the same as the diameter of the through hole 15, and the end portions of the flow paths 21 and 22 are opened upward. The volume of each reservoir is typically 10 to 1000 μl.
　***の分離は、第１の流路２１へ***を含む***サンプルを供給し、第２の流路２２へ媒体を供給し、合流流路２３において***サンプルと媒体とを層流状態で接触させることにより行う。層流状態で接触させることで、***サンプルに含まれる運動性の高い***（以下、運動性***という）は層流の界面から媒体流に入り込み、非運動性***は***サンプル流に残存する。こうして、運動性***が分離された***サンプルは第１の貯留部（廃棄液貯留部）４１に排出され、運動性***を含む媒体は第２の貯留部（運動性***貯留部）４２に排出され、運動性***と非運動性***とが分離される。ここで、媒体流の断面域を***サンプル流の断面域よりも大きくすることにより、分離効率を向上させることができる。なお、本実施例では、媒体を流通させる媒体流路を１つとしたが、複数形成してもよい。 In the separation of sperm, a semen sample containing sperm is supplied to the first flow path 21, a medium is supplied to the second flow path 22, and the semen sample and the medium are brought into contact in a laminar flow state in the merge flow path 23. By doing. By contacting in a laminar flow state, highly motile sperm contained in the semen sample (hereinafter referred to as motile sperm) enters the medium flow from the laminar flow interface, and non-motile sperm remains in the semen sample flow. Thus, the semen sample from which the motile sperm has been separated is discharged to the first reservoir (waste liquid reservoir) 41, and the medium containing the motile sperm is discharged to the second reservoir (motile sperm reservoir) 42. Motile sperm and non-motile sperm are separated. Here, the separation efficiency can be improved by making the cross-sectional area of the medium flow larger than the cross-sectional area of the semen sample flow. In this embodiment, one medium flow path for circulating the medium is used, but a plurality of medium flow paths may be formed.
　収容部本体２の側面には、４つの腕部３，３，３，３が、弾性変形可能に一体となって形成されている。収容部本体２のどの位置にどのような形状の腕部３をどの方向にどの程度形成するかは、適宜に設定されるが、本実施例では、収容部本体２の対向する側面それぞれの両端部から外方に凸の円弧形状の腕部３，３，３，３が延設され、同一側面から延設された腕部３，３が一つの円弧形状をなすように形成されている。腕部３は、収容部本体２を構成する上板１２の左右の側面１７，１８から上板１２の主面方向に延設されており、腕部３が収容部本体２の底面から離間して形成されている。例えば、収容部本体２の底面から１ｍｍ以上１５ｍｍ以下の高さに腕部３は形成される。 The four arm portions 3, 3, 3, and 3 are integrally formed on the side surface of the housing portion main body 2 so as to be elastically deformable. How much and in what direction the arm 3 is formed in which position in which position of the housing main body 2 is set as appropriate. In this embodiment, both ends of the opposing side surfaces of the housing main body 2 are arranged. The arc-shaped arm portions 3, 3, 3, and 3 that protrude outwardly from the portion extend, and the arm portions 3 and 3 that extend from the same side surface form a single arc shape. The arm portion 3 extends from the left and right side surfaces 17 and 18 of the upper plate 12 constituting the housing portion main body 2 in the main surface direction of the upper plate 12, and the arm portion 3 is separated from the bottom surface of the housing portion main body 2. Is formed. For example, the arm portion 3 is formed at a height of 1 mm or more and 15 mm or less from the bottom surface of the housing portion main body 2.
　図４の（ａ）から（ｆ）に、別の実施形態における腕部の形状を示す。（ａ），（ｄ）および（ｆ）では、腕部３の両端が収容部本体２に接合し、略半円形状もしくは略Ｖ字形状をなしている。（ｅ）および（ｆ）では、同一側面から延設された円弧状の腕部３，３は、その自由端が互いに反対方向を向くように形成されている。図４の（ｂ）では、円弧状の腕部３の先端部に外方に膨出した膨出部３ａが形成されている。 4 (a) to 4 (f) show the shape of the arm in another embodiment. In (a), (d), and (f), both ends of the arm portion 3 are joined to the housing portion main body 2 to form a substantially semicircular shape or a substantially V shape. In (e) and (f), the arc-shaped arm portions 3 and 3 extending from the same side surface are formed such that their free ends face in opposite directions. In FIG. 4B, a bulging portion 3 a bulging outward is formed at the tip of the arc-shaped arm portion 3.
　本実施例とは異なり、腕部は、バネ等で外方に付勢されたラッチ部材とされていてもよい。 Unlike the present embodiment, the arm portion may be a latch member biased outward by a spring or the like.
　収容部本体２および腕部３は、その材質を特に問わないが、成形性の観点から、熱可塑性樹脂が好ましい。熱可塑性樹脂としては、例えば、高密度ポリエチレン（ＨＤＰＥ）、中密度ポリエチレン（ＭＤＰＥ）、低密度ポリエチレン（ＬＤＰＥ）等のポリエチレン（ＰＥ）、ポリプロピレン（ＰＰ）、ポリ塩化ビニリデン、ポリ塩化ビニル（ＰＶＣ）、ポリスチレン（ＰＳ）、ポリ酢酸ビニル（ＰＶＡｃ）、テフロン（登録商標）などで知られるポリテトラフルオロエチレン（ＰＴＦＥ）、アクリロニトリルブタジエンスチレン樹脂（ＡＢＳ樹脂）、アクリロニトリルスチレン樹脂（ＡＳ樹脂）、ＰＭＭＡ等の（メタ）アクリル樹脂、ポリエチレンテレフタレート（ＰＥＴ）等の汎用プラスチック；ポリアミド（ＰＡ）、ポリアセタール（ＰＯＭ）、ポリカーボネート（ＰＣ）、変性ポリフェニレンエーテル（変性ＰＰＥ）、ポリブチレンテレフタレート（ＰＢＴ）、グラスファイバー強化ポリエチレンテレフタレート（ＧＦ－ＰＥＴ）、環状ポリオレフィン（ＣＯＰ）等のエンジニリングプラスチック；ポリフェニレンスルファイド（ＰＰＳ）、ポリテトラフルオロエチレン（ＰＴＦＥ）、ポリスルホン（ＰＳＦ）、ポリエーテルサルフォン（ＰＥＳ）、非晶ポリアリレート（ＰＡＲ）、液晶ポリマー（ＬＣＰ）、ポリエーテルエーテルケトン（ＰＥＥＫ）、熱可塑性ポリイミド（ＰＩ）、ポリアミドイミド（ＰＡＩ）等のスーパーエンジニアリングプラスチックが挙げられる。 The material of the housing part main body 2 and the arm part 3 is not particularly limited, but a thermoplastic resin is preferable from the viewpoint of moldability. Examples of the thermoplastic resin include polyethylene (PE) such as high density polyethylene (HDPE), medium density polyethylene (MDPE), and low density polyethylene (LDPE), polypropylene (PP), polyvinylidene chloride, and polyvinyl chloride (PVC). Polytetrafluoroethylene (PTFE) known as polystyrene (PS), polyvinyl acetate (PVAc), Teflon (registered trademark), acrylonitrile butadiene styrene resin (ABS resin), acrylonitrile styrene resin (AS resin), PMMA, etc. General purpose plastics such as (meth) acrylic resin, polyethylene terephthalate (PET); polyamide (PA), polyacetal (POM), polycarbonate (PC), modified polyphenylene ether (modified PPE), polybutylene terephthalate (PBT), glass fiber reinforced polyethylene terephthalate (GF-PET), engineering plastics such as cyclic polyolefin (COP); polyphenylene sulfide (PPS), polytetrafluoroethylene (PTFE), polysulfone (PSF), polyether Examples include super engineering plastics such as sulfone (PES), amorphous polyarylate (PAR), liquid crystal polymer (LCP), polyether ether ketone (PEEK), thermoplastic polyimide (PI), and polyamideimide (PAI).
　次に、本実施例の細胞取扱用具１の使用方法について説明する。図２および図３は、本実施例の細胞取扱用具１の使用状態を示す概略図である。細胞取扱用具１は、汎用のシャーレに代表される底部を有する円筒状容器１０１に載置され、円筒状容器１０１の内周面に弾性変形可能な腕部３の一部を当接させて固定されている。腕部３が収容部本体２の底面から離間して形成されているので、図示例のように円筒状容器１０１の内側底面の外周に沿って段差１０１ａが設けられていても、腕部３が段差１０１ａに当たることはない。このように固定された状態で、細胞の分離（選別）、培養、保存等を行う。 Next, a method of using the cell handling tool 1 of this example will be described. 2 and 3 are schematic views showing the usage state of the cell handling tool 1 of this embodiment. The cell handling tool 1 is placed on a cylindrical container 101 having a bottom typified by a general-purpose petri dish, and is fixed by bringing a part of an arm 3 that can be elastically deformed into contact with the inner peripheral surface of the cylindrical container 101. Has been. Since the arm portion 3 is formed away from the bottom surface of the housing portion main body 2, even if the step 101 a is provided along the outer periphery of the inner bottom surface of the cylindrical container 101 as in the illustrated example, the arm portion 3 is not It does not hit the step 101a. In such a fixed state, the cells are separated (selected), cultured, stored, and the like.
　円筒状容器１０１の蓋１０２をすれば、収容部本体２への菌の混入を防止できる。また、収容部本体２に収容される細胞への悪影響（例えば、溶媒が蒸発することによる影響）を抑制できる。 If the lid 102 of the cylindrical container 101 is used, it is possible to prevent the bacteria from being mixed into the housing body 2. Moreover, the bad influence (for example, influence by a solvent evaporating) to the cell accommodated in the accommodating part main body 2 can be suppressed.
　サンプルを管理する際、ラベルの貼付やＩＣチップの埋め込み等の処置が必要となるが、このような処置を細胞取扱用具１自体に施さず、収納容器１００に施せばよい。このように、サンプル管理機能を収納容器１００に付与することで、収容部本体２のサイズ変更、機能付与等のためのコストを削減することができる。細胞取扱用具１の廃棄は、容器１００に収納した状態で行えばよい。 When managing the sample, treatments such as labeling and IC chip embedding are required, but such treatments may be applied to the storage container 100 without being applied to the cell handling tool 1 itself. Thus, by providing the sample management function to the storage container 100, it is possible to reduce costs for changing the size of the storage unit main body 2 and for providing the function. What is necessary is just to perform the disposal of the cell handling tool 1 in the state accommodated in the container 100. FIG.
　上記実施例では、***分離用具として用いる場合について説明したが、例えば、収容部本体に培地を設けることにより細胞培養用具として適用可能である。 In the above-described embodiment, the case of using as a sperm separation tool has been described. However, for example, it can be applied as a cell culture tool by providing a medium in the container body.
　本発明の細胞取扱用具は、細胞の分離（選別）、培養、保存を行う器具として好適に用いられ得る。 The cell handling device of the present invention can be suitably used as a device for separating (selecting), culturing and storing cells.
　　　１　細胞取扱用具
　　　２　収容部本体
　　　３　腕部 1 Cell handling tool 2 Container body 3 Arm

Claims (6)

  1. 　細胞を収容し、底部が平板状の収容部本体と、
    　該収容部本体の外周面から外方に延設され、弾性を有する腕部と
    　を備える、細胞取扱用具。     Containing cells, and a container body having a flat bottom,
    A cell handling tool comprising: an elastic arm portion extending outward from an outer peripheral surface of the housing main body.
  2. 　前記腕部が外方に凸の円弧形状に形成されている、請求項１に記載の細胞取扱用具。 The cell handling tool according to claim 1, wherein the arm is formed in an outwardly convex arc shape.
  3. 　前記腕部が、前記収容部本体の底面から離間して設けられている、請求項１または２に記載の細胞取扱用具。 The cell handling tool according to claim 1 or 2, wherein the arm portion is provided apart from the bottom surface of the housing main body.
  4. 　底部を有する円筒状容器の内周面に前記腕部の一部を当接させて、前記収容部本体を該円筒状容器に固定させる、請求項１から３のいずれかに記載の細胞取扱用具。 The cell handling tool according to any one of claims 1 to 3, wherein a part of the arm portion is brought into contact with an inner peripheral surface of a cylindrical container having a bottom portion, and the container main body is fixed to the cylindrical container. .
  5. 　***分離用具として用いられる、請求項１から４のいずれかに記載の細胞取扱用具。 The cell handling tool according to any one of claims 1 to 4, which is used as a sperm separation tool.
  6. 　前記収容部本体は、***サンプルを流通させる第１の流路と、該第１の流路に***サンプルを供給する第１の供給部と、該第１の流路から排出される***サンプルを貯留する第１の貯留部と、媒体を流通させる第２の流路と、該第２の流路に媒体を供給する第２の供給部と、該第２の流路から排出される流体を貯留する第２の貯留部とを備え、
    　該第２の流路の中流部が該第１の流路との合流流路とされている、請求項５に記載の細胞取扱用具。     The container body includes a first flow path for circulating a semen sample, a first supply section for supplying the semen sample to the first flow path, and a semen sample discharged from the first flow path. A first storage section for storing, a second flow path for circulating the medium, a second supply section for supplying the medium to the second flow path, and a fluid discharged from the second flow path A second storage section for storing,
    The cell handling tool according to claim 5, wherein a midstream portion of the second flow path is a merge flow path with the first flow path.
PCT/JP2012/054900 2012-02-28 2012-02-28 Cell manipulation implement WO2013128561A1 (en)

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Cited By (1)

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JP2017531440A (en) * 2014-10-20 2017-10-26 ザ ユニバーシティ オブ ユタ リサーチ ファウンデイション Tissue sample processing system and related method

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JP2005518794A (en) * 2002-02-27 2005-06-30 ミシガン大学リージェンツ Method for selecting moving particles from low-moving particles and apparatus used therefor
JP2007515936A (en) * 2003-09-04 2007-06-21 アリックス インコーポレイテッド Particle and cell separation based on multilayer flow using laser steering
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JP2005518794A (en) * 2002-02-27 2005-06-30 ミシガン大学リージェンツ Method for selecting moving particles from low-moving particles and apparatus used therefor
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JP2012024020A (en) * 2010-07-23 2012-02-09 Nippi:Kk Culture vessel for storing slide glass

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