JP2008222640A

JP2008222640A - Collected tissue piece vitrification cryopreservation tool, tissue piece cryopreservation plate, and harvested tissue piece vitrification cryopreservation method

Info

Publication number: JP2008222640A
Application number: JP2007063978A
Authority: JP
Inventors: Noriko Kagawa; 則子香川; Masashige Kuwayama; 正成桑山; Futoshi Inoue; 太井上
Original assignee: KITAZATO SUPPLY CO Ltd
Current assignee: KITAZATO SUPPLY CO Ltd
Priority date: 2007-03-13
Filing date: 2007-03-13
Publication date: 2008-09-25
Anticipated expiration: 2027-03-13
Also published as: JP5224703B2

Abstract

【課題】組織片の採取が容易であり、採取した組織片を良好にガラス化凍結保存することが可能な採取組織片ガラス化凍結保存用具を提供するものである。
【解決手段】採取組織片ガラス化凍結保存用具１は、採取組織切取位置特定具１０と、採取組織切取位置特定具１０を用いて切り取られた組織片を凍結保存するための組織片凍結保存用プレート４とを備える。組織切取位置特定具１０は、平板状でありかつ所定幅の切取部特定用開口部２２と、裏面に設けられた切断具用係止部２３とを有する組織切取部特定部材２と、開口部２２を被包するように組織切取部特定部材２の表面側に押圧可能な開口部押圧部材３とを備える。凍結保存用プレート４は、切取部特定用開口部２２の所定幅と同じもしくは若干広い幅を有し、かつ所定長延び組織切取位置特定具１０を用いて切り取られた組織片を載置可能な金属性板状部材であり、多数の孔を備えている。
【選択図】図１An object of the present invention is to provide a vitrified cryopreservation tool for collecting tissue pieces that can be easily collected and vitrified and preserved well.
A harvested tissue piece vitrification cryopreservation tool 1 is a tissue slice cryopreservation tool 10 for cryopreserving a harvested tissue cutting position specifying tool 10 and a tissue piece cut using the collected tissue cutting position specifying tool 10. Plate 4. The tissue cutting position specifying tool 10 is a flat plate-shaped and has a predetermined width cutting part specifying opening 22 and a cutting tool specifying member 2 having a cutting tool locking part 23 provided on the back surface, and an opening. And an opening pressing member 3 that can be pressed on the surface side of the tissue cutout specifying member 2 so as to encapsulate 22. The cryopreservation plate 4 has a width that is the same as or slightly wider than the predetermined width of the cutout specifying opening 22, and can extend a predetermined length to mount a tissue piece cut out using the tissue cutout position specifying tool 10. It is a metallic plate-like member and has a large number of holes.
[Selection] Figure 1

Description

本発明は、哺乳動物の卵巣などの生体組織を凍結保存する際に使用する採取組織片ガラス化凍結保存用具、組織片凍結保存用プレートおよび採取組織片ガラス化凍結保存方法に関する。 The present invention relates to a harvested tissue piece vitrification cryopreservation tool, a tissue piece cryopreservation plate and a harvested tissue piece vitrification cryopreservation method used when cryopreserving a living tissue such as a mammal's ovary.

現在において、哺乳動物の生体細胞（例えば、精子、卵子など）の凍結保存が行われている。このような凍結保存は、特定の系統や品種の遺伝資源の保存を可能とする。また、絶滅の危機に瀕している動物種の維持にも有効である。さらに、ヒトの不妊治療においても有用である。
また、特開２００３−２８４５４６号公報（特許文献１）には、卵巣凍結法が開示されている。この卵巣凍結法は、非ヒト哺乳動物の雌個体から少なくとも卵巣を含む生殖組織を摘出し、これを１５〜０℃で保持した後にガラス化法により凍結することを特徴とするものである。
また、哺乳動物胚の凍結保存方法としては、特開２０００−１８９１５５公報（特許文献２）において、哺乳動物胚または卵子を滅菌処理した凍結ストロー、凍結バイアルまたは凍結チューブ等の凍結保存用容器の内面に、これらの胚または卵子を被包するに充分な最少量のガラス化液で貼り付け、この凍結保存用容器を密封し、そしてこの容器を液体窒素に接触させて急速に冷却することが提案されている。そして、融解方法では、前記の方法で保存した凍結保存用容器を前記液体窒素から取り出し、容器の一端部を開口し、この容器内に３３〜３９℃の希釈液を直接注入し、胚または卵子の凍結を融解希釈するものである。この方法によれば、哺乳動物胚または卵子をウイルスや細菌による感染のおそれがなく高い生存率で保存および融解希釈することのできるという優れた効果を備えている。
また、特開２００２−３１５５７３号公報（特許文献３）に示す卵凍結保存用具を本件出願人は提案している。この卵凍結保存用具は、耐寒性材料により形成された本体部と、本体部の一端に取り付けられ、可撓性かつ透明性かつ液体窒素耐性材料により形成された卵付着保持用ストリップと、該卵付着保持用ストリップを被包可能に本体部に着脱自在に取り付けられる一端が封鎖され、かつ耐寒性材料により形成された筒状部材とからなるものである。 Currently, cryopreservation of mammalian living cells (eg, sperm, eggs, etc.) is performed. Such cryopreservation enables the preservation of genetic resources of specific strains and varieties. It is also effective in maintaining endangered animal species. Furthermore, it is useful in the treatment of human infertility.
JP 2003-284546 A (Patent Document 1) discloses an ovary freezing method. This ovary freezing method is characterized in that a reproductive tissue including at least an ovary is extracted from a female individual of a non-human mammal, and this is kept at 15 to 0 ° C. and then frozen by vitrification.
In addition, as a cryopreservation method for mammalian embryos, in JP 2000-189155 A (Patent Document 2), the inner surface of a cryopreservation container such as a frozen straw, a cryovial, or a freezing tube obtained by sterilizing a mammalian embryo or egg. Proposed to paste with a minimum amount of vitrification solution sufficient to encapsulate these embryos or eggs, seal the cryopreservation container, and cool the container quickly by contacting it with liquid nitrogen Has been. In the thawing method, the cryopreservation container stored by the above method is taken out from the liquid nitrogen, one end of the container is opened, and a diluted solution of 33 to 39 ° C. is directly injected into the container, and an embryo or egg Thaw and dilute the freeze. This method has an excellent effect that mammalian embryos or ova can be stored and melt-diluted with a high survival rate without the risk of infection by viruses or bacteria.
In addition, the present applicant has proposed an egg cryopreservation tool disclosed in Japanese Patent Application Laid-Open No. 2002-315573 (Patent Document 3). The egg cryopreservation tool includes a main body portion formed of a cold-resistant material, an egg attachment holding strip attached to one end of the main body portion, formed of a flexible, transparent, and liquid nitrogen-resistant material, and the egg. One end removably attached to the main body so as to be able to enclose the adhesion holding strip is sealed, and is formed of a cylindrical member formed of a cold resistant material.

特開２００３−２８４５４６号公報JP 2003-284546 A 特開２０００−１８９１５５公報JP 2000-189155 A 特開２００２−３１５５７３号公報JP 2002-315573 A

上記の特許文献１のものでは、卵巣凍結法を開示するにすぎず、組織採取および組織凍結保存のための簡便な採取用具についての開示はない。また、上記の特許文献２および特許文献３のものも組織採取および組織凍結保存に利用できるものではない。 The thing of said patent document 1 only discloses the ovary freezing method, and there is no disclosure about a simple collection tool for tissue collection and tissue cryopreservation. Further, those described in Patent Document 2 and Patent Document 3 described above cannot be used for tissue collection and tissue cryopreservation.

そこで、本発明の目的は、組織片の採取が容易であり、かつ、採取した組織片を良好にガラス化凍結保存することが可能な採取組織片ガラス化凍結保存用具、組織片凍結保存用プレートおよび採取組織片ガラス化凍結保存方法を提供するものである。 Therefore, an object of the present invention is to collect a tissue piece vitrified cryopreservation tool, a tissue piece cryopreservation plate, which can easily collect the tissue piece and can be vitrified and preserved well. And a method for cryopreserving vitrified tissue pieces.

上記目的を達成するものは、以下のものである。
（１）採取組織切取位置特定具と、該採取組織切取位置特定具を用いて切り取られた組織片を凍結保存するための組織片凍結保存用プレートとを備える採取組織片ガラス化凍結保存用具であって、
前記採取組織切取位置特定具は、平板状でありかつ所定幅の切取部特定用開口部と、裏面に設けられ組織切断具の進行を係止するための切断具用係止部とを有する組織切取部特定部材と、前記切取部特定用開口部を被包するように前記組織切取部特定部材の表面側に押圧可能な開口部押圧部材とを備え、
前記組織片凍結保存用プレートは、前記切取部特定用開口部の所定幅と同じもしくは若干広い幅を有し、かつ所定長延び前記採取組織切取位置特定具を用いて切り取られた組織片を載置可能な金属性板状部材であるとともに、冷却媒体が侵入可能な多数の孔を備えている採取組織片ガラス化凍結保存用具。
（２）前記採取組織片ガラス化凍結保存用具は、卵巣組織片ガラス化凍結保存用具である上記（１）に記載の採取組織片ガラス化凍結保存用具。
（３）前記採取組織切取位置特定具は、前記組織切取部特定部材を対象生体組織に押圧した状態にて、前記切取部特定用開口部の内縁に沿って切り込みを入れた後、前記組織切取部特定部材の表面側に前記開口部押圧部材を前記切取部特定用開口部を被包するように押し当て前記切り込まれた組織部位を押圧した状態にて、前記組織切取部特定部材の裏面に沿って切断具を移動させることにより前記開口部の形状に対応した形状の組織片を切り取り可能である上記（１）または（２）に記載の採取組織片ガラス化凍結保存用具。 What achieves the above object is as follows.
(1) A harvested tissue piece vitrified cryopreservation tool comprising a harvested tissue cutout location specifying tool and a tissue piece cryopreservation plate for cryopreserving a tissue piece cut using the harvested tissue cutout location specifying tool. There,
The harvested tissue cutting position specifying tool has a flat plate shape and has a predetermined width cutting portion specifying opening and a cutting tool locking portion provided on the back surface for locking the progress of the tissue cutting tool. A cutout specifying member, and an opening pressing member that can be pressed on the surface side of the tissue cutout specifying member so as to encapsulate the cutout specifying opening,
The tissue piece cryopreservation plate has a width that is the same as or slightly wider than the predetermined width of the cutout specifying opening, and extends a predetermined length and mounts the cut tissue piece cut using the collected tissue cutout position specifying tool. A harvested tissue piece vitrified cryopreservation tool which is a metal plate-like member that can be placed and has a large number of holes through which a cooling medium can enter.
(2) The harvested tissue piece vitrified cryopreservation tool according to (1) above, wherein the harvested tissue piece vitrified cryopreservation tool is an ovarian tissue vitrification cryopreservation tool.
(3) The tissue cut-out position specifying tool is configured to cut the tissue cut-out along the inner edge of the cut-out specifying opening in a state where the tissue cut-out specifying member is pressed against the target biological tissue. The back surface of the tissue cutout specifying member is pressed against the cutout portion specifying member by pressing the opening pressing member against the front side of the cutout specifying member and enclosing the cutout specifying opening. The harvested tissue piece vitrified cryopreservation device according to (1) or (2) above, wherein a tissue piece having a shape corresponding to the shape of the opening can be cut by moving the cutting tool along the cutting edge.

（４）前記切取部特定用開口部は、矩形状であり、前記組織片凍結保存用プレートは、前記矩形状の前記開口部の幅と同じもしくは若干広い幅を有し、かつ、前記矩形状の前記開口部の長さより十分に長い長さを有するものである上記（１）ないし（３）のいずれかに記載の採取組織片ガラス化凍結保存用具。
（５）前記切取部特定用開口部は、円もしくは楕円状であり、前記組織片凍結保存用プレートは、前記開口部の直径または短軸と同じもしくは若干広い幅を有し、かつ、前記開口部の直径および長軸の長さより十分に長い長さを有するものである上記（１）ないし（３）のいずれかに記載の採取組織片ガラス化凍結保存用具。
（６）前記切断具用係止部は、前記切取部特定用開口部より前記組織切取部特定部材の端部側に設けられている上記（１）ないし（５）のいずれかに記載の採取組織片ガラス化凍結保存用具。
（７）前記切断具用係止部は、前記切取部特定用開口部付近に設けられている上記（１）ないし（６）のいずれかに記載の採取組織片ガラス化凍結保存用具。
（８）前記開口部押圧部材は、透明平板状部材である上記（１）ないし（７）のいずれかに記載の採取組織片ガラス化凍結保存用具。
（９）前記組織片凍結保存用プレートは、略矩形状の薄板プレートであり、全体にわたり冷却媒体が侵入可能な多数の孔を有している上記（１）ないし（８）のいずれかに記載の採取組織片ガラス化凍結保存用具。
（１０）前記組織片凍結保存用プレートは、銅、銅合金、アルミ、アルミ合金、金、金合金、銀、銀合金から選択された金属により形成されている上記（１）ないし（９）のいずれかに記載の採取組織片ガラス化凍結保存用具。
（１１）前記組織切取部特定部材は、表面の両側部に一端側より他端側に延びるように設けられるとともに、前記開口部押圧部材の両側部をスライド可能に収納する開口部押圧部材誘導部を備えている上記（１）ないし（１０）のいずれかに記載の採取組織片ガラス化凍結保存用具。
（１２）前記組織切取部特定部材は、裏面の少なくとも一方の側部に一端側より他端側に延びるように設けられるとともに、前記切断具がスライド可能な切断具誘導部を備えている上記（１）ないし（１１）のいずれかに記載の採取組織片ガラス化凍結保存用具。
（１３）前記切断具誘導部の先端部は、前記組織切取部特定部材の一端より突出している上記（１２）に記載の採取組織片ガラス化凍結保存用具。 (4) The cutout specifying opening has a rectangular shape, and the tissue piece cryopreservation plate has a width the same as or slightly wider than the rectangular opening, and the rectangular shape. The collected tissue piece vitrified cryopreservation device according to any one of (1) to (3), which has a length sufficiently longer than the length of the opening.
(5) The cutout specifying opening is a circle or an ellipse, and the tissue piece cryopreservation plate has the same or slightly wider width than the diameter or short axis of the opening, and the opening The harvested tissue piece vitrified cryopreservation device according to any one of the above (1) to (3), which has a length sufficiently longer than the diameter of the part and the length of the long axis.
(6) The sampling according to any one of (1) to (5), wherein the cutting tool locking portion is provided on an end side of the tissue cutout specifying member from the cutout specifying opening. Tissue piece vitrification cryopreservation tool.
(7) The harvested tissue piece vitrified cryopreservation device according to any one of (1) to (6), wherein the cutting tool locking portion is provided in the vicinity of the cutout specifying opening.
(8) The harvested tissue piece vitrified cryopreservation device according to any one of (1) to (7), wherein the opening pressing member is a transparent flat plate member.
(9) The tissue piece cryopreservation plate is a substantially rectangular thin plate, and has a large number of holes through which a cooling medium can invade, as described in any one of (1) to (8). Collected tissue piece vitrification cryopreservation tool.
(10) The tissue piece cryopreservation plate is formed of a metal selected from copper, copper alloy, aluminum, aluminum alloy, gold, gold alloy, silver, and silver alloy. The harvested tissue piece vitrified cryopreservation device according to any one of the above.
(11) The tissue cutting part specifying member is provided on both sides of the surface so as to extend from one end side to the other end side, and the opening pressing member guiding part that slidably accommodates both side parts of the opening pressing member. The harvested tissue piece vitrified cryopreservation device according to any one of (1) to (10), comprising:
(12) The tissue cut portion specifying member is provided on at least one side portion of the back surface so as to extend from one end side to the other end side, and further includes a cutting tool guide portion on which the cutting tool can slide. 1) The harvested tissue piece vitrified cryopreservation device according to any one of 1) to (11).
(13) The collected tissue piece vitrified cryopreservation device according to (12), wherein a distal end portion of the cutting tool guiding portion protrudes from one end of the tissue cutting portion specifying member.

また、上記目的を達成するものは、以下のものである。
（１４）所定長延びる金属性板状部材であり、組織片載置部を有するとともに、少なくとも組織片載置部には、冷却媒体が侵入可能な多数の孔を有する組織片凍結保存用プレート。
（１５）前記組織片凍結保存用プレートは、銅、銅合金、アルミ、アルミ合金、金、金合金、銀、銀合金から選択された金属により形成されている上記（１４）に記載の組織片凍結保存用プレート。
（１６）前記組織片凍結保存用プレートは、略矩形状の薄板プレートであり、全体にわたり冷却媒体が侵入可能な多数の孔を有している上記（１４）または（１５）に記載のの組織片凍結保存用プレート。 Moreover, what achieves the said objective is as follows.
(14) A tissue piece cryopreservation plate which is a metallic plate-like member extending for a predetermined length, has a tissue piece placement section, and has a plurality of holes through which a cooling medium can enter at least in the tissue piece placement section.
(15) The tissue piece according to (14), wherein the tissue piece cryopreservation plate is formed of a metal selected from copper, copper alloy, aluminum, aluminum alloy, gold, gold alloy, silver, and silver alloy. Plate for cryopreservation.
(16) The tissue piece cryopreservation plate is a substantially rectangular thin plate, and the tissue according to (14) or (15), which has a large number of holes through which a cooling medium can enter. Single cryopreservation plate.

また、上記目的を達成するものは、以下のものである。
（１７）上記（１）ないし（１３）のいずれかに記載の採取組織片ガラス化凍結保存用具を用いる採取組織片ガラス化凍結保存方法であって、
前記組織切取部特定部材を対象生体組織に押圧した状態にて、前記切取部特定用開口部の内縁に沿って切り込みを入れた後、前記組織切取部特定部材の表面側に前記開口部押圧部材を前記切取部特定用開口部を被包するように押し当て前記切り込まれた組織部位を押圧した状態にて、前記組織切取部特定部材の裏面に沿って切断具を移動させることにより前記開口部の形状に対応した形状の組織片を切り取る組織片採取工程と、採取した組織片をガラス化液に浸漬する組織片ガラス化液浸漬工程と、ガラス化液より前記組織片を取り出し、前記組織片凍結保存用プレート上に載置する組織片載置工程と、組織片が載置された組織片凍結保存用プレートを冷却媒体タンク内に投入する投入工程とを行う採取組織片ガラス化凍結保存方法。
（１８）前記採取組織片ガラス化凍結保存方法は、卵巣組織片ガラス化凍結保存方法もしくは肝臓組織片ガラス化凍結保存方法である上記（１７）に記載の採取組織片ガラス化凍結保存方法。 Moreover, what achieves the said objective is as follows.
(17) A harvested tissue piece vitrified cryopreservation method using the harvested tissue piece vitrified cryopreservation device according to any of (1) to (13) above,
After making a cut along the inner edge of the opening for specifying the cut-out portion in a state where the tissue cut-out specifying member is pressed against the target living tissue, the opening pressing member on the surface side of the tissue cut-out specifying member The opening is moved by moving the cutting tool along the back surface of the tissue cutout specifying member in a state where the cut tissue specifying portion is pressed against the cutout specifying opening. A tissue piece collecting step of cutting out a tissue piece having a shape corresponding to the shape of the part, a tissue piece vitrifying solution immersing step of immersing the collected tissue piece in a vitrification solution, and taking out the tissue piece from the vitrification solution, the tissue Collected tissue piece vitrified cryopreservation that performs a tissue piece placement step of placing on a piece cryopreservation plate and a loading step of placing the tissue piece cryopreservation plate on which the tissue piece is placed into a cooling medium tank Method.
(18) The collected tissue piece vitrified cryopreservation method according to (17), wherein the collected tissue piece vitrified cryopreservation method is an ovarian tissue vitrification cryopreservation method or a liver tissue vitrification cryopreservation method.

本発明の採取組織片ガラス化凍結保存用具は、採取組織切取位置特定具と、該採取組織切取位置特定具を用いて切り取られた組織片を凍結保存するための組織片凍結保存用プレートとを備える。そして、採取組織切取位置特定具は、平板状でありかつ所定幅の切取部特定用開口部と裏面に設けられた切断具用係止部とを有する組織切取部特定部材と、前記切取部特定用開口部を被包するように前記組織切取部特定部材の表面側に押圧可能な開口部押圧部材とを備えるので、的確な大きさおよび厚さの組織片を採取できる。さらに、組織片凍結保存用プレートは、切取部特定用開口部の所定幅と同じもしくは若干広い幅を有し、かつ所定長延び前記採取組織切取位置特定具を用いて切り取られた組織片を載置可能な金属性板状部材であるとともに、冷却媒体が侵入可能な多数の孔を備えているので、採取した組織片をガラス化液に浸漬した後、この組織片凍結保存用プレート上に載置し、冷却媒体タンク内に投入することにより、良好にガラス化状態にて凍結させることができる。 The harvested tissue piece vitrification cryopreservation tool of the present invention comprises a harvested tissue cutting position specifying tool, and a tissue piece cryopreservation plate for cryopreserving a tissue piece cut using the harvested tissue cutting position specifying tool. Prepare. Then, the collected tissue cutting position specifying tool has a flat plate shape and has a predetermined width cutting part specifying opening and a cutting tool locking member provided on the back surface, and the cutting part specifying member Since the opening pressing member that can be pressed on the surface side of the tissue cutout specifying member is provided so as to encapsulate the opening for use, a tissue piece of an accurate size and thickness can be collected. Furthermore, the tissue piece cryopreservation plate has a width that is the same as or slightly wider than the predetermined width of the opening for specifying the cut-out portion, and extends a predetermined length to mount the tissue piece cut out using the collected tissue cut-out position specifying tool. Since it is a metal plate-like member that can be placed and has many holes through which the cooling medium can enter, the collected tissue piece is immersed in the vitrification solution and then placed on this tissue piece cryopreservation plate. It can be frozen in a vitrified state by placing it in the cooling medium tank.

また、本発明の組織片凍結保存用プレートは、所定長延びる金属性板状部材であり、組織片載置部を有するとともに、少なくとも組織片載置部には、冷却媒体が侵入可能な多数の孔を有するものである。このため、採取した組織片をガラス化液に浸漬した後、この組織片凍結保存用プレート上に載置し、冷却媒体タンク内に投入することにより、良好にガラス化状態にて凍結させることができる。 The tissue piece cryopreservation plate of the present invention is a metal plate-like member extending a predetermined length, and has a tissue piece placement section, and at least a large number of cooling media can enter the tissue piece placement section. It has a hole. For this reason, after immersing the collected tissue piece in a vitrification solution, it is placed on this tissue piece cryopreservation plate and put into a cooling medium tank, so that it can be frozen well in a vitrified state. it can.

また、本発明の採取組織片ガラス化凍結保存方法は、上記の本発明の採取組織片ガラス化凍結保存用具を用いるものであり、さらに、組織切取部特定部材を対象生体組織に押圧した状態にて、切取部特定用開口部の内縁に沿って切り込みを入れた後、組織切取部特定部材の表面側に開口部押圧部材を押圧して切り込まれた組織部位を押圧した状態にて、組織切取部特定部材の裏面に沿って切断具を移動させることにより開口部の形状に対応した形状の組織片を切り取る組織片採取工程と、採取した組織片をガラス化液に浸漬する組織片ガラス化液浸漬工程と、ガラス化液より前記組織片を取り出し、組織片凍結保存用プレート上に載置する組織片載置工程と、組織片が載置された組織片凍結保存用プレートを冷却媒体タンク内に投入する投入工程とを行うものであるので、的確な大きさおよび厚さの組織片を、良好にガラス化状態にて凍結させることができる。 The harvested tissue piece vitrification cryopreservation method of the present invention uses the above-described harvested tissue piece vitrification cryopreservation tool of the present invention, and further presses the tissue cutout part specifying member against the target biological tissue. Then, after cutting along the inner edge of the cutout specifying opening, pressing the opening pressing member on the surface side of the tissue cutout specifying member, pressing the cut tissue portion, the tissue A tissue piece collecting step of cutting a tissue piece having a shape corresponding to the shape of the opening by moving the cutting tool along the back surface of the cutout specific member, and vitrification of the tissue piece in which the collected tissue piece is immersed in a vitrification solution A liquid immersion step, a tissue piece placing step of taking out the tissue piece from the vitrification solution and placing it on the tissue piece cryopreservation plate, and a tissue piece cryopreservation plate on which the tissue piece is placed on the cooling medium tank Input Since it performs a degree, the tissue pieces of exact size and thickness, good can be frozen at vitrified state.

本発明の採取組織片ガラス化凍結保存用具および採取組織片ガラス化凍結保存方法を図面に示した実施例を用いて説明する。
図１は、本発明の採取組織片ガラス化凍結保存用具の一実施例の正面図である。
図２は、図１に示した採取組織片ガラス化凍結保存用具の採取組織切取位置特定具の背面図である。図３は、図１のＡ−Ａ線断面図である。図４は、図１のＢ−Ｂ線断面図である。図５は、図１のＣ−Ｃ線断面図である。 The harvested tissue piece vitrified cryopreservation tool and the harvested tissue piece vitrified cryopreservation method of the present invention will be described with reference to the embodiments shown in the drawings.
FIG. 1 is a front view of one embodiment of a harvested tissue piece vitrification cryopreservation device of the present invention.
FIG. 2 is a rear view of the collected tissue cutting position specifying tool of the collected tissue piece vitrification cryopreservation tool shown in FIG. 3 is a cross-sectional view taken along line AA in FIG. 4 is a cross-sectional view taken along line BB in FIG. 5 is a cross-sectional view taken along the line CC of FIG.

本発明の採取組織片ガラス化凍結保存用具１は、採取組織切取位置特定具１０と、採取組織切取位置特定具１０を用いて切り取られた組織片を凍結保存するための組織片凍結保存用プレート４とを備える。
そして、採取組織切取位置特定具１０は、平板状でありかつ所定幅の切取部特定用開口部２２と、裏面に設けられた切断具用係止部２３とを有する組織切取部特定部材２と、切取部特定用開口部２２を被包するように組織切取部特定部材２の表面側に押圧可能な開口部押圧部材３とを備える。そして、組織片凍結保存用プレート４は、切取部特定用開口部２２の所定幅と同じもしくは若干広い幅を有し、かつ所定長延び採取組織切取位置特定具１０を用いて切り取られた組織片を載置可能な金属性板状部材であるとともに、冷却媒体が侵入可能な多数の孔を備えている。 The harvested tissue piece vitrification cryopreservation tool 1 of the present invention includes a harvested tissue cutting position specifying tool 10 and a tissue piece cryopreservation plate for cryopreserving a tissue piece cut using the harvested tissue cutting position specifying tool 10. 4.
And the extract | collecting tissue cutting position specific tool 10 is flat shape, and the structure cutting part specific member 2 which has the opening part 22 for cutting part specification of predetermined width, and the latching | locking part 23 for cutting tools provided in the back surface, And an opening pressing member 3 that can be pressed on the surface side of the tissue cutout specifying member 2 so as to encapsulate the cutout specifying opening 22. The tissue piece cryopreservation plate 4 has a width that is the same as or slightly wider than the predetermined width of the cutout specifying opening 22 and extends a predetermined length, and is cut out using the collected tissue cutout position specifying tool 10. And a large number of holes through which the cooling medium can enter.

また、本発明の組織片凍結保存用プレート４は、所定長延びる金属性板状部材であり、組織片載置部を有するとともに、少なくとも組織片載置部には、冷却媒体が侵入可能な多数の孔を有するものである。
そして、本発明の採取組織片ガラス化凍結保存用具および組織片凍結保存用プレートは、牛、ヒトなどの各種の生体組織の採取組織片ガラス化凍結保存に使用できる。特に、卵巣組織片をガラス化凍結保存するのに好適である。
また、本発明の採取組織片ガラス化凍結保存用具１では、組織切取部特定部材２を対象生体組織に押圧した状態にて、切取部特定用開口部２２の内縁に沿って切り込みを入れた後、組織切取部特定部材２の表面側に開口部押圧部材３を切取部特定用開口部２２を被包するように押し当て切り込まれた組織部位を押圧した状態にて、組織切取部特定部材２の裏面に沿って切断具を移動させることにより開口部２２の形状に対応した形状の組織片を切り取ることが可能なものとなっている。 Further, the tissue piece cryopreservation plate 4 of the present invention is a metal plate-like member extending a predetermined length, and has a tissue piece placement portion, and a large number of cooling media can enter at least the tissue piece placement portion. It has a hole.
The harvested tissue piece vitrified cryopreservation tool and the tissue piece cryopreservation plate of the present invention can be used for harvested tissue piece vitrified cryopreservation of various biological tissues such as cattle and humans. In particular, it is suitable for preserving vitrified ovarian tissue fragments.
Further, in the harvested tissue piece vitrification cryopreservation tool 1 of the present invention, after cutting the tissue cutout specifying member 2 along the inner edge of the cutout specifying opening 22 in a state where the tissue cutout specifying member 2 is pressed against the target biological tissue. The tissue cutting part specifying member is pressed in a state in which the opening pressing member 3 is pressed against the surface side of the tissue cutting part specifying member 2 so as to enclose the opening 22 for specifying the cutting part and the tissue part pressed is cut. The tissue piece having a shape corresponding to the shape of the opening 22 can be cut out by moving the cutting tool along the back surface of 2.

本発明の採取組織片ガラス化凍結保存用具１は、図１に示すように、採取組織切取位置特定具１０と、採取組織切取位置特定具１０を用いて切り取られた組織片を凍結保存するための組織片凍結保存用プレート４とを備える。なお、この実施例では、採取組織片ガラス化凍結保存用具１は、採取組織切取位置特定具１０と、組織片凍結保存用プレート４とのセットとなっているが、本発明の組織片凍結保存用プレート４は、採取組織切取位置特定具１０を用いることなく採取された組織片にも使用できる。
採取組織切取位置特定具１０は、組織切取部特定部材２と、開口部押圧部材３とを備える。 As shown in FIG. 1, the harvested tissue piece vitrification cryopreservation tool 1 of the present invention is for cryopreserving a harvested tissue cutting position specifying tool 10 and a tissue piece cut using the harvested tissue cutting position specifying tool 10. The tissue piece cryopreservation plate 4 is provided. In this embodiment, the harvested tissue piece vitrification cryopreservation tool 1 is a set of the harvested tissue cutting position specifying tool 10 and the tissue piece cryopreservation plate 4, but the tissue piece cryopreservation of the present invention. The plate 4 can be used for a tissue piece collected without using the collected tissue cutting position specifying tool 10.
The collected tissue cutting position specifying tool 10 includes a tissue cutting portion specifying member 2 and an opening pressing member 3.

組織切取部特定部材２は、図１ないし図３に示すように、略矩形状に形成された平板状部材であり、表面および裏面は、平坦となっている。そして、組織切取部特定部材２の平面部２１の中央部には、所定幅かつ所定長延びる矩形状の切取部特定用開口部２２が設けられている。
この実施例では、切取部特定用開口部２２は、矩形状のものとなっており、後述する組織片凍結保存用プレート４は、矩形状の開口部２２の幅と同じもしくは若干広い幅を有し、かつ、矩形状の開口部２２の長さより十分に長い長さを有している。 As shown in FIGS. 1 to 3, the tissue cutout specifying member 2 is a flat plate member formed in a substantially rectangular shape, and the front surface and the back surface are flat. A rectangular cutout specifying opening 22 having a predetermined width and a predetermined length is provided at the center of the flat portion 21 of the tissue cutout specifying member 2.
In this embodiment, the cutout specifying opening 22 has a rectangular shape, and the later-described tissue piece cryopreservation plate 4 has a width that is the same as or slightly wider than the width of the rectangular opening 22. In addition, the length is sufficiently longer than the length of the rectangular opening 22.

また、組織切取部特定部材２は、裏面側であり、切取部特定用開口部２２付近でありかつ切取部特定用開口部２２よりどちらかの端部側に設けられた切断具用係止部２３を有している。このため、組織切取部特定部材２の裏面は、切断具用係止部２３部分が突出した状態となっており、その他の部分は平坦面となっている。特に、この実施例では、切断具用係止部２３は、組織切取部特定部材２の他端と切取部特定用開口部２２間であり、かつ切取部特定用開口部２２と近接する位置に設けられている。切断具用係止部２３は、切取部特定用開口部２２付近に設けられていることが好ましい。
組織切取部特定部材２の大きさとしては、矩形状の場合、長片が３０〜５０ｍｍ程度、短辺が、２０〜４０ｍｍ程度であることが好ましい。また、組織切取部特定部材２の厚さとしては、０．５〜２ｍｍ程度が好ましい。また、切断具用係止部２３の厚さとしては、０．５〜２ｍｍ程度が好ましい。また、切取部特定用開口部２２の大きさとしては、２５〜２５０ｍｍ^２程度が好ましく、矩形状の場合、各辺の長さが、５〜１５ｍｍ程度であることが好ましい。 Further, the tissue cutout part specifying member 2 is on the back surface side, near the cutout part specifying opening 22 and on either end side of the cutout part specifying opening 22. 23. For this reason, the back surface of the tissue cut-out part specifying member 2 is in a state in which the cutting tool locking part 23 portion protrudes, and the other part is a flat surface. In particular, in this embodiment, the cutting tool locking portion 23 is located between the other end of the tissue cutout specifying member 2 and the cutout specifying opening 22 and close to the cutout specifying opening 22. Is provided. The cutting tool locking portion 23 is preferably provided in the vicinity of the cut portion specifying opening 22.
In the case of a rectangular shape, the size of the tissue cutout specifying member 2 is preferably about 30 to 50 mm for the long piece and about 20 to 40 mm for the short side. Moreover, as thickness of the structure | tissue cutting part specific member 2, about 0.5-2 mm is preferable. Moreover, as thickness of the latching | locking part 23 for cutting tools, about 0.5-2 mm is preferable. Moreover, as a magnitude | size of the opening part 22 for cutting part specification, about 25-250 mm < ² > is preferable, and when it is rectangular shape, it is preferable that the length of each side is about 5-15 mm.

また、組織切取部特定部材２の形成材料としては、透明もしくは半透明であることが好ましく、さらに、生体組織付着性を持たないものが望ましい。組織切取部特定部材２の形成材料としては、ポリオレフィン（例えば、ポリエチレンポリプロピレン、エチレン−プロピレン共重合体、エチレン−酢酸ビニル共重合体）、スチレン系樹脂（例えば、ポリスチレン、メタクリレート−スチレン共重合体、メタクリレート−ブチレン−スチレン共重合体）、ポリエステル（例えば、ポリエチレンテレフタレート、ポリブチレンテレフタレート）、ポリアミド（例えば、６ナイロン，６６ナイロン）、ポリテトラフロロエチレン、エチレン−テトラフロロエチレン共重合体、テトラフロロエチレン−ヘキサフロロプロピレン共重合体、塩化三フッ化エチレン樹脂などのフッ素樹脂等が好適に使用される。 In addition, the material for forming the tissue cutout specifying member 2 is preferably transparent or translucent, and more preferably has no biological tissue adhesion. As a material for forming the tissue cut-out part specifying member 2, polyolefin (for example, polyethylene polypropylene, ethylene-propylene copolymer, ethylene-vinyl acetate copolymer), styrene resin (for example, polystyrene, methacrylate-styrene copolymer, (Methacrylate-butylene-styrene copolymer), polyester (for example, polyethylene terephthalate, polybutylene terephthalate), polyamide (for example, 6 nylon, 66 nylon), polytetrafluoroethylene, ethylene-tetrafluoroethylene copolymer, tetrafluoroethylene -A fluororesin such as a hexafluoropropylene copolymer or a chloroethylene trifluoride resin is preferably used.

開口部押圧部材３は、透明平板状部材が用いられており、この実施例では、組織切取部特定部材２の形状に対応した矩形状のものとなっている。そして、開口部押圧部材３は、切取部特定用開口部２２を被包するように組織切取部特定部材２の表面側に押圧可能である。この実施例のように、開口部押圧部材３は、組織切取部特定部材２の外形形状に対応した形状とすることが好ましい。なお、開口部押圧部材３は、組織切取部特定部材２より若干小さいものまた若干大きいものであってもよい。
開口部押圧部材３の大きさとしては、矩形状の場合、長片が３０〜５０ｍｍ程度、短辺が、２０〜４０ｍｍ程度であることが好ましい。また、開口部押圧部材３の厚さとしては、０．５〜２ｍｍ程度が好ましい。また、開口部押圧部材３の形成材料としては、上述した組織切取部特定部材２において説明したものが好適に使用できる。 The opening pressing member 3 is a transparent flat plate member. In this embodiment, the opening pressing member 3 has a rectangular shape corresponding to the shape of the tissue cutout specifying member 2. And the opening part press member 3 can be pressed to the surface side of the structure | tissue cut part specific member 2 so that the opening part 22 for cut part specific | specification may be covered. As in this embodiment, it is preferable that the opening pressing member 3 has a shape corresponding to the outer shape of the tissue cutout specifying member 2. Note that the opening pressing member 3 may be slightly smaller or slightly larger than the tissue cutout specifying member 2.
In the case of a rectangular shape, the size of the opening pressing member 3 is preferably about 30 to 50 mm for the long piece and about 20 to 40 mm for the short side. Moreover, as thickness of the opening part press member 3, about 0.5-2 mm is preferable. Moreover, as a forming material of the opening part pressing member 3, what was demonstrated in the structure | tissue cutting part specific member 2 mentioned above can be used conveniently.

そして、組織片凍結保存用プレート４は、切取部特定用開口部２２の所定幅と同じもしくは若干広い幅を有し、かつ所定長延び採取組織切取位置特定具１０を用いて切り取られた組織片を載置可能な平板状のものとなっている。
この実施例では、組織片凍結保存用プレート４は、切取部特定用開口部２２の一辺の長さと実質的に同じ幅を有する細長い矩形状のものとなっている。組織片凍結保存用プレート４の大きさとしては、矩形状の場合、長片が５０〜１５０ｍｍ程度、短辺が、５〜５０ｍｍ程度であることが好ましい。
そして、組織片凍結保存用プレート４は、冷却媒体が侵入可能な多数の孔４２を備えている。このように、組織片凍結保存用プレート４が、冷却媒体が侵入可能な多数の孔４２を有することにより、プレート自体の冷却が早いとともに、組織片のプレート載置側にも冷却媒体が接触可能であるため、組織片のプレート載置側からの冷却も良好となる。この両者の効果により、組織片の冷却が極めて迅速に行われる。また、組織切取部特定部材２および開口部押圧部材３を用いることにより、組織片凍結保存用プレート４の形状に対応するとともに、的確な厚さの組織片を採取することができる。これらの作用が複合することにより、ガラス化凍結保存後の解凍時における細胞の高い生存率を得ることができ、自家移植に極めて有効である。 The tissue piece cryopreservation plate 4 has a width that is the same as or slightly wider than the predetermined width of the cutout specifying opening 22 and extends a predetermined length, and is cut out using the collected tissue cutout position specifying tool 10. Is a flat plate that can be mounted.
In this embodiment, the tissue piece cryopreservation plate 4 has an elongated rectangular shape having a width substantially the same as the length of one side of the cutout specifying opening 22. In the case of a rectangular shape, the size of the tissue piece cryopreservation plate 4 is preferably about 50 to 150 mm for the long piece and about 5 to 50 mm for the short side.
The tissue piece cryopreservation plate 4 includes a large number of holes 42 into which the cooling medium can enter. As described above, the tissue piece cryopreservation plate 4 has a large number of holes 42 through which the cooling medium can enter, so that the plate itself can be cooled quickly and the cooling medium can also contact the plate mounting side of the tissue piece. Therefore, the cooling of the tissue piece from the plate placement side is also good. Due to both effects, the tissue piece is cooled very quickly. In addition, by using the tissue cutout specifying member 2 and the opening pressing member 3, it is possible to collect a tissue piece having an appropriate thickness while corresponding to the shape of the tissue piece cryopreservation plate 4. By combining these actions, a high cell survival rate upon thawing after vitrification cryopreservation can be obtained, which is extremely effective for autologous transplantation.

この実施例では、組織片凍結保存用プレート４は、図１に示すように、略矩形状の薄板プレートであり、全体にわたり冷却媒体が侵入可能な多数の孔を有している。孔の形状としては、図１に示すような円形のものが好ましい。孔の大きさとしては、２〜８ｍｍ^２程度が好ましく、特に、４〜７ｍｍ^２程度が好ましい。また、組織片凍結保存用プレート４の全体面積に対する孔の総面積は、３０〜７０％程度が好ましい。組織片凍結保存用プレート４は、金属性板状部材により形成されている。用いられる金属としては、特に制限されないが、銅、銅合金、アルミ、アルミ合金、金、金合金、銀、銀合金などが好適である。組織片凍結保存用プレート４は、液体窒素浸漬時における１分間換算冷却速度が−２万℃を越えるものであることが好ましい。このような冷却速度を有することにより、組織片を良好にガラス化状態にて凍結することができる。 In this embodiment, the tissue piece cryopreservation plate 4 is a substantially rectangular thin plate as shown in FIG. 1, and has a large number of holes through which the cooling medium can invade. As the shape of the hole, a circular shape as shown in FIG. 1 is preferable. The size of the pores is preferably about 2 to 8 mm ^2, in particular, about 4 to 7 mm ² is preferred. The total area of the holes with respect to the entire area of the tissue piece cryopreservation plate 4 is preferably about 30 to 70%. The tissue piece cryopreservation plate 4 is formed of a metal plate member. Although it does not restrict | limit especially as a metal used, Copper, a copper alloy, aluminum, an aluminum alloy, gold | metal | money, a gold alloy, silver, a silver alloy etc. are suitable. The tissue piece cryopreservation plate 4 preferably has a cooling rate equivalent to 1 minute when immersed in liquid nitrogen exceeds −20,000 ° C. By having such a cooling rate, the tissue piece can be frozen in a vitrified state well.

なお、組織片凍結保存用プレートに形成される孔の形状は、図１に示すような円形タイプのものに限定されるものではなく、図２２に示す組織片凍結保存用プレート４ａのような、多角形形状（この実施例では、四角形）の孔４２ａであってもよい。
さらに、組織片凍結保存用プレートに形成される孔は、図２３に示すように、組織片が載置される領域にのみ形成されていてもよい。この実施例の組織片凍結保存用プレート４ｂでは、一端側の所定領域にのみ孔４２ｂが設けられている。孔４２ｂが形成される領域のプレートの長手方向の長さとしては、切取部特定用開口部２２の長さの２〜５倍程度であることが好ましい。孔４２ｂの大きさとしては、上述したものと同様のものが好ましく、さらには、図２２に示したような、多角形形状の孔であってもよい。 In addition, the shape of the hole formed in the tissue piece cryopreservation plate is not limited to the circular type as shown in FIG. 1, but like the tissue piece cryopreservation plate 4a shown in FIG. The hole 42a having a polygonal shape (in this embodiment, a quadrangle) may be used.
Further, the hole formed in the tissue piece cryopreservation plate may be formed only in a region where the tissue piece is placed, as shown in FIG. In the tissue piece cryopreservation plate 4b of this embodiment, a hole 42b is provided only in a predetermined region on one end side. The length of the plate in the longitudinal direction of the region in which the hole 42b is formed is preferably about 2 to 5 times the length of the cutout specifying opening 22. The size of the hole 42b is preferably the same as described above, and may be a polygonal hole as shown in FIG.

次に、本発明の他の実施例について説明する。
図６ないし図８は、本発明の他の実施例の採取組織片ガラス化凍結保存用具に使用される組織切取部特定部材の背面図である。
組織切取部特定部材２ａの角部は、図６の実施例の組織切取部特定部材２ａに示すように、面取りされた状態となっていてもよい。また、切断具用係止部の形態は、図１ないし図３に示したものに限定されるものではなく、図６に示す切断具用係止部２３ａ、２３ｂのように、組織切取部特定部材２ａの他端と切取部特定用開口部２２間であり、かつ、組織切取部特定部材２ａの両側部に設けてもよい。また、切断具用係止部２３の形状は、図６に示すように、円形状であってもよく、また、楕円形状、多角形状などであってもよい。 Next, another embodiment of the present invention will be described.
6 to 8 are rear views of the tissue cutout specifying member used in the harvested tissue piece vitrification cryopreservation device of another embodiment of the present invention.
The corners of the tissue cutout specifying member 2a may be chamfered as shown in the tissue cutout specifying member 2a in the embodiment of FIG. Further, the form of the cutting tool locking portion is not limited to that shown in FIGS. 1 to 3, and the tissue cutting portion specifying portion is specified like the cutting tool locking portions 23 a and 23 b shown in FIG. 6. It may be provided between the other end of the member 2a and the cutout specifying opening 22 and on both sides of the tissue cutout specifying member 2a. Further, the shape of the cutting tool locking portion 23 may be circular as shown in FIG. 6, or may be elliptical, polygonal, or the like.

また、採取組織切取位置特定具の切取部特定用開口部の形状は、図１ないし図３に示したものに限定されるものではなく、図７の実施例の組織切取部特定部材２ｂの開口部２２ａのように、円形状もしく楕円形状であってもよい。そして、この実施例においても、図７に示すように、組織切取部特定部材２ｂの他端と切取部特定用開口部２２ａ間に切断具用係止部２３ｃが設けられている。
そして、このようなタイプの組織切取部特定部材２ｂが用いられる場合、組織片凍結保存用プレートとしては、開口部２２ａの直径または短軸と同じもしくは若干広い幅を有し、かつ、開口部２２ａの直径または長軸の長さより十分に長い長さを有するものが用いられる。 Further, the shape of the cutout specifying opening of the harvested tissue cutout position specifying tool is not limited to that shown in FIGS. 1 to 3, but the opening of the tissue cutout specifying member 2b in the embodiment of FIG. Like the part 22a, it may be circular or elliptical. Also in this embodiment, as shown in FIG. 7, a cutting tool locking portion 23c is provided between the other end of the tissue cut portion specifying member 2b and the cut portion specifying opening 22a.
When such a type of tissue cutout specifying member 2b is used, the tissue piece cryopreservation plate has the same or slightly wider width than the diameter or short axis of the opening 22a, and the opening 22a. Those having a length sufficiently longer than the diameter or the length of the major axis are used.

さらに、図８の実施例の組織切取部特定部材２ｃに示すように、切取部特定用開口部２２ａの形状が、円形状もしくは楕円形状であるとともに、切断具用係止部２３ａ、２３ｂが、組織切取部特定部材２ａの他端と切取部特定用開口部２２ａ間であり、かつ、組織切取部特定部材２ｃの両側部に設けられたものであってもよい。そして、切断具用係止部２３ａ，２３ｂの形状は、円形状、楕円形状、多角形状などであってもよい。 Furthermore, as shown in the tissue cutting part specifying member 2c of the embodiment of FIG. 8, the shape of the cutting part specifying opening 22a is circular or elliptical, and the locking parts 23a and 23b for the cutting tool are It may be provided between the other end of the tissue cutout specifying member 2a and the cutout specifying opening 22a and on both sides of the tissue cutout specifying member 2c. And the shape of the latching | locking parts 23a, 23b for cutting tools may be circular shape, elliptical shape, polygonal shape, etc.

また、上述した全ての実施例の採取組織切取位置特定具は、図９ないし図１１に示すような開口部押圧部材誘導部２４ａ，２４ｂを備えていてもよい。
図９は、本発明の他の実施例の採取組織片ガラス化凍結保存用具に使用される組織切取部特定部材の正面図である。図１０は、図９のＤ−Ｄ線断面図である。図１１は、図９に示した実施例の採取組織片ガラス化凍結保存用具の作用を説明するための説明図である。
この実施例における組織切取部特定部材２ｄは、表面の両側部に一端側より他端側に延びるように設けられるとともに、開口部押圧部材３の両側部をスライド可能に収納する開口部押圧部材誘導部２４ａ，２４ｂを備えている。開口部押圧部材誘導部２４ａ，２４ｂは、組織切取部特定部材２ｄの一端側より他端側に延びている。特に、この実施例では、開口部押圧部材誘導部２４ａ，２４ｂは、組織切取部特定部材２ｄの一端より他端までの全長に渡り形成されている。開口部押圧部材誘導部２４ａ，２４ｂは、図１０に示すように、開口部押圧部材３の両側部を収納する凹部２５ａ，２５ｂを備えており、この凹部２５ａ，２５ｂ内に開口部押圧部材３の両側部が収納されるとともに、摺動可能となっている。また、開口部押圧部材誘導部２４ａ，２４ｂは、凹部２５ａ，２５ｂの上方を被包しており、組織切取部特定部材２ｄからの開口部押圧部材３の離脱を規制している。 In addition, the collected tissue cutting position specifying tools of all the embodiments described above may include opening portion pressing member guiding portions 24a and 24b as shown in FIGS.
FIG. 9 is a front view of a tissue cutout specifying member used for a harvested tissue piece vitrification cryopreservation tool according to another embodiment of the present invention. 10 is a cross-sectional view taken along the line DD of FIG. FIG. 11 is an explanatory diagram for explaining the operation of the harvested tissue piece vitrification cryopreservation tool of the embodiment shown in FIG. 9.
The tissue cutout specifying member 2d in this embodiment is provided on both sides of the surface so as to extend from one end side to the other end side, and the opening pressing member guide for slidably storing both side portions of the opening pressing member 3 Parts 24a and 24b are provided. The opening pressing member guiding portions 24a and 24b extend from the one end side of the tissue cutout specifying member 2d to the other end side. In particular, in this embodiment, the opening pressing member guiding portions 24a and 24b are formed over the entire length from one end to the other end of the tissue cut portion specifying member 2d. As shown in FIG. 10, the opening portion pressing member guiding portions 24a and 24b are provided with concave portions 25a and 25b for accommodating both side portions of the opening portion pressing member 3, and the opening portion pressing member 3 is provided in the concave portions 25a and 25b. The both side portions are accommodated and slidable. In addition, the opening pressing member guiding portions 24a and 24b enclose the recesses 25a and 25b, and restrict the detachment of the opening pressing member 3 from the tissue cutout specifying member 2d.

この実施例の採取組織切取位置特定具では、図１１に示すように、開口部押圧部材３の端部が、組織切取部特定部材２ｄの開口部押圧部材誘導部２４ａ，２４ｂに侵入し、かつ、切取部特定用開口部２２を開口部押圧部材３が被包しない状態とする。そして、この状態にて、摘出された生体器官（例えば、卵巣、肝臓）に、採取組織切取位置特定具を押しつけ、切取部特定用開口部２２の内縁に沿って、切断具にて切り込みを入れる。これにより、切り込みにより囲まれた生体器官部位が、開口部内に侵入する。続いて、開口部押圧部材３をスライドさせて、切取部特定用開口部２２が開口部押圧部材３にて被包された状態とする。この実施例のように、開口部押圧部材誘導部２４ａ，２４ｂを設けることにより、開口部押圧部材３の切取部特定用開口部２２上への配置作業が容易なものとなる。
そして、開口部押圧部材３を組織切取部特定部材２ｄに押圧し、その状態を維持したまま、組織切取部特定部材２ｄの下面に沿って、切断具を切断具用係止部２３方向に移動させる。これにより、切取部特定用開口部２２の形状に対応した組織片が切断される。 In the harvested tissue cutting position specifying tool of this embodiment, as shown in FIG. 11, the end of the opening pressing member 3 enters the opening pressing member guiding portions 24a and 24b of the tissue cutting portion specifying member 2d, and The opening specifying member 22 is not covered by the opening pressing member 3. In this state, the extracted tissue cutting position specifying tool is pressed against the extracted living organ (for example, ovary, liver), and the cutting tool is cut along the inner edge of the cutting portion specifying opening 22. . As a result, the living organ part surrounded by the cut enters the opening. Subsequently, the opening pressing member 3 is slid so that the cutout specifying opening 22 is encapsulated by the opening pressing member 3. By providing the opening pressing member guiding portions 24a and 24b as in this embodiment, the arrangement work of the opening pressing member 3 on the cutout specifying opening 22 is facilitated.
Then, the opening pressing member 3 is pressed against the tissue cutting part specifying member 2d, and the cutting tool is moved in the direction of the cutting tool locking part 23 along the lower surface of the tissue cutting part specifying member 2d while maintaining the state. Let Thereby, the tissue piece corresponding to the shape of the cutout specifying opening 22 is cut.

さらに、上述した全ての実施例の採取組織切取位置特定具は、図１２ないし図１７に示すような構成を有するものであってもよい。
図１２は、本発明の他の実施例の採取組織片ガラス化凍結保存用具に使用される採取組織切取位置特定具の組織切取部特定部材の正面図である。図１３は、図１２に示した採取組織切取位置特定具に使用される開口部押圧部材の正面図である。図１４は、図１２のＥ−Ｅ線断面図である。図１５は、図１２のＦ−Ｆ線断面図である。図１６および図１７は、図１２ないし図１５に示した実施例の採取組織片ガラス化凍結保存用具の作用を説明するための説明図である。 Furthermore, the collected tissue cutting position specifying tools of all the embodiments described above may have a configuration as shown in FIGS.
FIG. 12 is a front view of a tissue cutting portion specifying member of a sampling tissue cutting position specifying tool used in a sampling tissue vitrification cryopreservation tool according to another embodiment of the present invention. FIG. 13 is a front view of an opening pressing member used in the collected tissue cutting position specifying tool shown in FIG. 14 is a cross-sectional view taken along line EE in FIG. 15 is a cross-sectional view taken along line FF in FIG. 16 and 17 are explanatory views for explaining the operation of the harvested tissue piece vitrification cryopreservation device of the embodiment shown in FIGS. 12 to 15.

この実施例における組織切取部特定部材２ｅは、上述した組織切取部特定部材２ｄと同様に、表面の両側部に一端側より他端側に延びるように設けられるとともに、開口部押圧部材３の両側部をスライド可能に収納する開口部押圧部材誘導部２４ａ，２４ｂを備えている。開口部押圧部材誘導部２４ａ，２４ｂは、組織切取部特定部材２ｄの一端側より他端側に延びている。特に、この実施例では、開口部押圧部材誘導部２４ａ，２４ｂは、組織切取部特定部材２ｅの一端より他端までの全長に渡り形成されている。
開口部押圧部材誘導部２４ａ，２４ｂは、図１２，図１４および図１５に示すように、開口部押圧部材３ａの両側部を収納する凹部２５ａ，２５ｂと、組織切取部特定部材２ｅの表面側より凹部２５ａ，２５ｂに延びる段部２６ａ、２６ｂを備えている。さらに、各段部２６ａ、２６ｂの一方の端部には、所定長延びる段部欠損部２７ａ，２７ｂが形成されている。また、図１３に示すように、開口部押圧部材３ａの一方の端部には、両側部に突出部３１ａ，３１ｂを有することにより、組織切取部特定部材２ｅの段部２６ａと段部２６ｂ間の距離より広い幅を持ちかつ段部欠損部２７ａと段部欠損部２７ｂ間の距離より狭い幅を有し、所定長延びる幅広部３１と、この幅広部３１と連続するとともに、両側部に切欠部３２ａ，３２ｂを有することにより、幅が狭くなった幅狭部３２を備えている。幅狭部３２の幅は、組織切取部特定部材２ｅの段部２６ａと段部２６ｂ間の距離より短いものとなっている。そして、凹部２５ａ，２５ｂ内に開口部押圧部材３の幅広部３１の端部が収納されるとともに、凹部２５ａ，２５ｂ内かつ段部２６ａ、２６ｂ上を摺動可能となっている。また、開口部押圧部材誘導部２４ａ，２４ｂは、凹部２５ａ，２５ｂの上方を被包しており、組織切取部特定部材２ｅからの開口部押圧部材３ａの離脱を規制している。 The tissue cutout part specifying member 2e in this embodiment is provided on both sides of the surface so as to extend from one end side to the other end side as well as the tissue cutout part specifying member 2d described above, and both sides of the opening pressing member 3 Opening portion pressing member guiding portions 24a and 24b for slidably storing the portion. The opening pressing member guiding portions 24a and 24b extend from the one end side of the tissue cutout specifying member 2d to the other end side. In particular, in this embodiment, the opening pressing member guiding portions 24a and 24b are formed over the entire length from one end to the other end of the tissue cutout specifying member 2e.
As shown in FIGS. 12, 14, and 15, the opening pressing member guiding portions 24 a and 24 b include concave portions 25 a and 25 b that house both side portions of the opening pressing member 3 a and the surface side of the tissue cutout specifying member 2 e. Further, step portions 26a and 26b extending to the recesses 25a and 25b are provided. Furthermore, step missing portions 27a and 27b extending a predetermined length are formed at one end of each step 26a and 26b. Further, as shown in FIG. 13, one end portion of the opening pressing member 3 a has projecting portions 31 a and 31 b on both side portions, so that the step 26 a and the step portion 26 b of the tissue cut portion specifying member 2 e are provided. And a width that is narrower than the distance between the stepped portion 27a and the stepped portion 27b. The wide portion 31 extends for a predetermined length, is continuous with the wide portion 31, and is notched on both sides. By having the portions 32a and 32b, a narrow portion 32 having a narrow width is provided. The width of the narrow portion 32 is shorter than the distance between the step portion 26a and the step portion 26b of the tissue cutout specifying member 2e. And the edge part of the wide part 31 of the opening part press member 3 is accommodated in recessed part 25a, 25b, and it can slide in the recessed part 25a, 25b and on the step part 26a, 26b. In addition, the opening pressing member guiding portions 24a and 24b enclose the concave portions 25a and 25b, and restrict the detachment of the opening pressing member 3a from the tissue cutout specifying member 2e.

この実施例の採取組織切取位置特定具では、図１６に示すように、開口部押圧部材３ａの幅広部３１が、組織切取部特定部材２ｅの開口部押圧部材誘導部２４ａ，２４ｂに侵入し、かつ、切取部特定用開口部２２を開口部押圧部材３ａが被包しない状態とする。そして、この状態にて、摘出された生体器官（例えば、卵巣、肝臓）に、採取組織切取位置特定具を押しつけ、切取部特定用開口部２２の内縁に沿って、切断具にて切り込みを入れる。これにより、切り込みにより囲まれた生体器官部位が、開口部内に侵入する。続いて、開口部押圧部材３ａをスライドさせて、切取部特定用開口部２２が開口部押圧部材３ａにて被包された状態とする。この実施例では、開口部押圧部材３ａは、組織切取部特定部材２ｅの表面に実質的に接触することなく、段部２６ａ、２６ｂ上をスライドする。このため、開口部押圧部材３ａは、開口部２２内に侵入した生体器官部位にも接触することなくスライドする。 In the harvested tissue cutting position specifying tool of this embodiment, as shown in FIG. 16, the wide portion 31 of the opening pressing member 3a enters the opening pressing member guiding portions 24a and 24b of the tissue cutting portion specifying member 2e, And let the opening part 22a for cutting part specification not be encapsulated by the opening part press member 3a. In this state, the extracted tissue cutting position specifying tool is pressed against the extracted living organ (for example, ovary, liver), and the cutting tool is cut along the inner edge of the cutting portion specifying opening 22. . As a result, the living organ part surrounded by the cut enters the opening. Subsequently, the opening pressing member 3a is slid so that the cutout specifying opening 22 is encapsulated by the opening pressing member 3a. In this embodiment, the opening pressing member 3a slides on the stepped portions 26a and 26b without substantially contacting the surface of the tissue cutout specifying member 2e. Therefore, the opening pressing member 3 a slides without contacting the living organ part that has entered the opening 22.

そして、開口部押圧部材３ａのスライドを進行させ、開口部押圧部材３ａの幅広部３１の全体が、組織切取部特定部材２ｅの段部欠損部２７ａ，２７ｂに到達すると、図１７に示すように、幅広部３１は、組織切取部特定部材２ｅの段部欠損部２７ａ，２７ｂ内に侵入可能となり、開口部押圧部材３ａを組織切取部特定部材２ｅ側に押圧可能となる。そして、開口部押圧部材３ａを組織切取部特定部材２ｅに押圧し、その状態を維持したまま、組織切取部特定部材２ｅの下面に沿って、切断具を切断具用係止部２３方向に移動させる。これにより、切取部特定用開口部２２の形状に対応した組織片が切断される。 Then, when the sliding of the opening pressing member 3a is advanced and the entire wide portion 31 of the opening pressing member 3a reaches the stepped portions 27a and 27b of the tissue cutout specifying member 2e, as shown in FIG. The wide portion 31 can enter the stepped portion 27a, 27b of the tissue cutout specifying member 2e, and can press the opening pressing member 3a toward the tissue cutout specifying member 2e. Then, the opening pressing member 3a is pressed against the tissue cutting part specifying member 2e, and the cutting tool is moved in the direction of the cutting tool locking part 23 along the lower surface of the tissue cutting part specifying member 2e while maintaining the state. Let Thereby, the tissue piece corresponding to the shape of the cutout specifying opening 22 is cut.

さらに、上述した全ての実施例の採取組織切取位置特定具は、図１８ないし図２１に示すような構成を有するものであってもよい。
図１８は、本発明の他の実施例の採取組織片ガラス化凍結保存用具に使用される採取組織切取位置特定具の組織切取部特定部材の背面図である。図１９は、図１８に示した組織切取部特定部材の拡大平面図である。図２０は、図１８のＧ−Ｇ線拡大断面図である。図２１は、図１８および図１９に示した実施例の採取組織片ガラス化凍結保存用具の作用を説明するための説明図である。
この実施例の組織切取部特定部材２ｆは、裏面の少なくとも一方の側部に一端側より他端側に延びるように設けられるとともに、切断具がスライド可能な切断具誘導部を備えている。このような誘導部を備えることにより、摘出された生体器官より組織片の切断がより容易なものとなる。特に、この実施例では、裏面の両側部に一端側より他端側に延びるように設けられるとともに、切断具がスライド可能な切断具誘導部５１ａ、５１ｂを備えている。そして、この実施例では、切断具誘導部５１ａ、５１ｂの先端部は、組織切取部特定部材２ｆの一端より突出し、切断具の誘導を容易なものとしている。 Furthermore, the collected tissue cutting position specifying tools of all the embodiments described above may have a configuration as shown in FIGS.
FIG. 18 is a rear view of a tissue cutting portion specifying member of a sampling tissue cutting position specifying tool used in a sampling tissue vitrification cryopreservation tool according to another embodiment of the present invention. FIG. 19 is an enlarged plan view of the tissue cutout specifying member shown in FIG. 20 is an enlarged cross-sectional view taken along line GG in FIG. FIG. 21 is an explanatory diagram for explaining the operation of the harvested tissue piece vitrification cryopreservation device of the example shown in FIGS. 18 and 19.
The tissue cut portion specifying member 2f of this embodiment is provided on at least one side portion of the back surface so as to extend from one end side to the other end side, and includes a cutting tool guiding portion on which the cutting tool can slide. By providing such a guide part, it becomes easier to cut the tissue piece than the extracted living organ. In particular, in this embodiment, cutting tool guiding portions 51a and 51b are provided on both sides of the back surface so as to extend from one end side to the other end side, and the cutting tool can slide. And in this Example, the front-end | tip part of the cutting tool guidance | induction part 51a, 51b protrudes from the end of the tissue cutting part specific member 2f, and makes it easy to guide | invade a cutting tool.

この実施例の組織切取部特定部材２ｆでは、切断具誘導部５１ａ、５１ｂは、所定長延びる平板状のものとなっており、図１８ないし図２０に示すように、先端部が自由端となっており、基端部５２ａ，５２ｂが、組織切取部特定部材２ｆの切断具用係止部２３の形成部位に対応する側部に固定されている。さらに、切断具誘導部５１ａ、５１ｂは、組織切取部特定部材２ｆの裏面に実質的に接触せず、両者間には、間隙５３ａ，５３ｂが形成されている。そして、切断具誘導部５１ａ、５１ｂの先端部は、組織切取部特定部材２ｆの一端より突出している。
この実施例の採取組織切取位置特定部材２ｆを摘出された生体器官（例えば、卵巣、肝臓）に、採取組織切取位置特定具を押しつけ、切取部特定用開口部２２の内縁に沿って、切断具にて切り込みを入れる。これにより、切り込みにより囲まれた生体器官部位が、開口部内に侵入する。続いて、開口部押圧部材３を組織切取部特定部材２ｆに押圧し、その状態を維持したまま、組織切取部特定部材２ｆの一端より突出している切断具誘導部５１ａ、５１ｂ上に切断具７を載置する。そして、切断具７を切断具誘導部５１ａ、５１ｂを切断具用係止部２３方向に移動させると、図２１（裏面側から見た図）に示すように、切断具７は、切断具誘導部５１ａ、５１ｂと組織切取部特定部材２ｆの裏面間に挟まれた状態にて、両者間を進行することにより、切取部特定用開口部２２の形状に対応した組織片が切断される。 In the tissue cut portion specifying member 2f of this embodiment, the cutting tool guide portions 51a and 51b are flat plate-like shapes extending a predetermined length, and the tip portion is a free end as shown in FIGS. The base end portions 52a and 52b are fixed to the side portions corresponding to the formation site of the cutting tool locking portion 23 of the tissue cutout portion specifying member 2f. Further, the cutting tool guiding portions 51a and 51b do not substantially contact the back surface of the tissue cut portion specifying member 2f, and gaps 53a and 53b are formed between them. And the front-end | tip part of the cutting tool induction | guidance | derivation part 51a, 51b protrudes from the end of the tissue cutting part specific member 2f.
The collected tissue cutting position specifying member is pressed against a living organ (for example, ovary, liver) from which the collected tissue cutting position specifying member 2f of this embodiment is extracted, and the cutting tool is cut along the inner edge of the cut portion specifying opening 22 Make a cut at. As a result, the living organ part surrounded by the cut enters the opening. Subsequently, the opening pressing member 3 is pressed against the tissue cutting part specifying member 2f, and the cutting tool 7 is placed on the cutting tool guiding parts 51a and 51b protruding from one end of the tissue cutting part specifying member 2f while maintaining the state. Is placed. Then, when the cutting tool 7 is moved in the direction of the cutting tool locking portion 23, the cutting tool 7 is guided to the cutting tool 7 as shown in FIG. 21 (viewed from the back side). A tissue piece corresponding to the shape of the cutout specifying opening 22 is cut by advancing between the portions 51a and 51b and the backside of the tissue cutout specifying member 2f.

次に、本発明の採取組織片ガラス化凍結保存方法について説明する。
本発明の採取組織片ガラス化凍結保存方法は、上述した採取組織片ガラス化凍結保存用具を用いるものであり、使用する採取組織片ガラス化凍結保存用具としては、上述したいずれの実施例のものを用いてもよい。
本発明の採取組織片ガラス化凍結保存方法は、組織切取部特定部材を対象生体組織（例えば、摘出された生体器官）に押圧した状態にて、切取部特定用開口部の内縁に沿って切り込みを入れた後、組織切取部特定部材の表面側に開口部押圧部材を押圧して前記切り込まれた組織部位を押圧した状態にて、組織切取部特定部材の裏面に沿って切断具を移動させることにより、開口部の形状に対応した形状の組織片を切り取る組織片採取工程と、採取した組織片をガラス化液に浸漬する組織片ガラス化液浸漬工程と、ガラス化液より組織片を取り出し、組織片凍結保存用プレート上に載置する組織片載置工程と、組織片が載置された組織片凍結保存用プレートを冷却媒体タンク内に投入する投入工程とを行うものである。 Next, the method for cryopreserving the collected tissue piece vitrified according to the present invention will be described.
The harvested tissue piece vitrification cryopreservation method of the present invention uses the above-described harvested tissue piece vitrification cryopreservation tool, and the harvested tissue piece vitrification cryopreservation tool used is that of any of the above-described examples. May be used.
The method for cryopreserving a collected tissue piece vitrified according to the present invention cuts along the inner edge of the cutout specifying opening in a state where the tissue cutout specifying member is pressed against a target biological tissue (for example, a removed living organ). After cutting, move the cutting tool along the back surface of the tissue cutout specifying member while pressing the opening pressing member on the surface side of the tissue cutout specifying member and pressing the cut tissue part. A tissue piece collecting step of cutting a tissue piece having a shape corresponding to the shape of the opening, a tissue piece vitrifying solution immersing step of immersing the collected tissue piece in a vitrification solution, and a tissue piece from the vitrification solution The tissue piece placing step of taking out and placing on the tissue piece cryopreservation plate and the charging step of putting the tissue piece cryopreservation plate on which the tissue piece is placed into the cooling medium tank are performed.

そして、本発明の採取組織片ガラス化凍結保存方法は、卵巣組織片ガラス化凍結保存方法もしくは肝臓組織片ガラス化凍結保存方法に用いることが有効である。
組織片採取工程については、上述した採取組織片ガラス化凍結保存用具において説明した通りである。
組織片ガラス化液浸漬工程は、採取した組織片をガラス化液に浸漬することにより行われる。 The harvested tissue piece vitrification cryopreservation method of the present invention is effective for use in the ovarian tissue vitrification cryopreservation method or the liver tissue vitrification cryopreservation method.
The tissue piece collection step is as described in the above-described collected tissue piece vitrification cryopreservation tool.
The tissue piece vitrification solution immersion step is performed by immersing the collected tissue piece in a vitrification solution.

ガラス化液としては、従来より用いられているものを使用することができる。具体的には、グリセロール、プロピレングリコール、ジメチルスルホサイド（DMSO）、エチレングリコール、ブタンジオールなどから選択される１種または２種以上の細胞膜透過性凍結保護物質と、シュークロース、トレハロース、パーコール、ポリエチレングリコール、ポリヴィニルピロリドン、ウシ血清アルブミン、フィコールなどから選択される１種または２種以上の細胞膜非透過性凍結保護物質を含有するものが好適である。
また、ガラス化液に浸漬する前に、採取した組織片の平衡液浸漬処理を行うことが好ましい。平衡液としては、細胞膜透過性凍結保護物質の低濃度溶液が好ましい。細胞膜透過性凍結保護物質としては、上述したものが使用できる。
そして、組織片ガラス化液浸漬工程が終了した後、ガラス化液より組織片を取り出し、組織片凍結保存用プレート４上に載置する組織片載置工程が行われる。組織片は、凍結保存用プレート４の孔４２が存在する部分に載置する。
そして、組織片が載置された組織片凍結保存用プレートを冷却媒体タンク内に投入する投入工程が行われる。冷却媒体としては、液体窒素が好適である。 As the vitrification liquid, those conventionally used can be used. Specifically, one or more cell membrane-permeable cryoprotectants selected from glycerol, propylene glycol, dimethylsulfoside (DMSO), ethylene glycol, butanediol and the like, sucrose, trehalose, percoll, polyethylene Those containing one or more cell membrane-impermeable cryoprotectants selected from glycol, polyvinyl pyrrolidone, bovine serum albumin, Ficoll and the like are suitable.
Moreover, it is preferable to perform the equilibration liquid immersion process of the extract | collected tissue piece before immersing in vitrification liquid. The equilibrium solution is preferably a low concentration solution of a cell membrane-permeable cryoprotectant. As the cell membrane-permeable cryoprotectant, those described above can be used.
Then, after the tissue piece vitrification solution immersion step is completed, a tissue piece placement step is performed in which the tissue piece is taken out of the vitrification solution and placed on the tissue piece cryopreservation plate 4. The tissue piece is placed on the portion of the cryopreservation plate 4 where the holes 42 are present.
Then, a charging step of charging the tissue piece cryopreservation plate on which the tissue piece is placed into the cooling medium tank is performed. As the cooling medium, liquid nitrogen is suitable.

図１は、本発明の採取組織片ガラス化凍結保存用具の一実施例の正面図である。FIG. 1 is a front view of one embodiment of a harvested tissue piece vitrification cryopreservation device of the present invention. 図２は、図１に示した採取組織片ガラス化凍結保存用具の組織切取部特定部材の背面図である。FIG. 2 is a rear view of the tissue cutout specifying member of the harvested tissue piece vitrification cryopreservation tool shown in FIG. 1. 図３は、図１のＡ−Ａ線断面図である。3 is a cross-sectional view taken along line AA in FIG. 図４は、図１のＢ−Ｂ線断面図である。4 is a cross-sectional view taken along line BB in FIG. 図５は、図１のＣ−Ｃ線断面図である。5 is a cross-sectional view taken along the line CC of FIG. 図６は、本発明の他の実施例の採取組織片ガラス化凍結保存用具に使用される組織切取部特定部材の背面図である。FIG. 6 is a rear view of the tissue cutout specifying member used in the harvested tissue piece vitrification cryopreservation tool of another embodiment of the present invention. 図７は、本発明の他の実施例の採取組織片ガラス化凍結保存用具に使用される組織切取部特定部材の背面図である。FIG. 7 is a rear view of the tissue cutout specifying member used in the harvested tissue piece vitrification cryopreservation tool of another embodiment of the present invention. 図８は、本発明の他の実施例の採取組織片ガラス化凍結保存用具に使用される組織切取部特定部材の背面図である。FIG. 8 is a rear view of the tissue cutout specifying member used in the harvested tissue piece vitrification cryopreservation device of another embodiment of the present invention. 図９は、本発明の他の実施例の採取組織片ガラス化凍結保存用具に使用される組織切取部特定部材の正面図である。FIG. 9 is a front view of a tissue cutout specifying member used for a harvested tissue piece vitrification cryopreservation tool according to another embodiment of the present invention. 図１０は、図９のＤ−Ｄ線断面図である。10 is a cross-sectional view taken along the line DD of FIG. 図１１は、図９に示した実施例の採取組織片ガラス化凍結保存用具の作用を説明するための説明図である。FIG. 11 is an explanatory diagram for explaining the operation of the harvested tissue piece vitrification cryopreservation tool of the embodiment shown in FIG. 9. 図１２は、本発明の他の実施例の採取組織片ガラス化凍結保存用具に使用される組織切取部特定部材の正面図である。FIG. 12 is a front view of a tissue cutout specifying member used in a harvested tissue piece vitrification cryopreservation tool according to another embodiment of the present invention. 図１３は、本発明の他の実施例の採取組織片ガラス化凍結保存用具に使用される開口部押圧部材の正面図である。FIG. 13: is a front view of the opening part press member used for the extraction | collection tissue piece vitrification cryopreservation tool of the other Example of this invention. 図１４は、図１２のＥ−Ｅ線断面図である。14 is a cross-sectional view taken along line EE in FIG. 図１５は、図１２のＦ−Ｆ線断面図である。15 is a cross-sectional view taken along line FF in FIG. 図１６は、図１２ないし図１４に示した実施例の採取組織片ガラス化凍結保存用具の作用を説明するための説明図である。FIG. 16 is an explanatory diagram for explaining the operation of the harvested tissue piece vitrification cryopreservation device of the embodiment shown in FIGS. 12 to 14. 図１７は、図１２ないし図１４に示した実施例の採取組織片ガラス化凍結保存用具の作用を説明するための説明図である。FIG. 17 is an explanatory diagram for explaining the operation of the harvested tissue piece vitrification cryopreservation device of the embodiment shown in FIGS. 12 to 14. 図１８は、本発明の他の実施例の採取組織片ガラス化凍結保存用具に使用される採組織切取部特定部材の背面図である。FIG. 18 is a rear view of a tissue-collecting-cutting-part specifying member used for a harvested tissue piece vitrification cryopreservation device according to another embodiment of the present invention. 図１９は、図１８に示した採取組織切取位置特定具の拡大平面図である。FIG. 19 is an enlarged plan view of the collected tissue cutting position specifying tool shown in FIG. 図２０は、図１８のＧ−Ｇ線拡大断面図である。20 is an enlarged cross-sectional view taken along line GG in FIG. 図２１は、図１８および図１９に示した実施例の採取組織片ガラス化凍結保存用具の作用を説明するための説明図である。FIG. 21 is an explanatory diagram for explaining the operation of the harvested tissue piece vitrification cryopreservation device of the example shown in FIGS. 18 and 19. 図２２は、本発明の他の実施例の採取組織片ガラス化凍結保存用具に使用される組織片凍結保存用プレートの正面図である。FIG. 22 is a front view of a tissue piece cryopreservation plate used in a harvested tissue piece vitrification cryopreservation device according to another embodiment of the present invention. 図２３は、本発明の他の実施例の採取組織片ガラス化凍結保存用具に使用される組織片凍結保存用プレートの正面図である。FIG. 23 is a front view of a tissue piece cryopreservation plate used in a harvested tissue piece vitrification cryopreservation device according to another embodiment of the present invention.

Explanation of symbols

１採取組織片ガラス化凍結保存用具
２組織切取部特定部材
３開口部押圧部材
４組織片凍結保存用プレート
１０採取組織切取位置特定具 DESCRIPTION OF SYMBOLS 1 Collected tissue piece vitrification cryopreservation tool 2 Tissue cut part specifying member 3 Opening part pressing member 4 Tissue piece cryopreservation plate 10 Collected tissue cut position specifying tool

Claims

A harvested tissue piece vitrified cryopreservation tool comprising a harvested tissue cutting position specifying tool and a tissue piece cryopreservation plate for cryopreserving a tissue piece cut using the collected tissue cutting position specifying tool,
The harvested tissue cutting position specifying tool has a flat plate shape and has a predetermined width cutting portion specifying opening and a cutting tool locking portion provided on the back surface for locking the progress of the tissue cutting tool. A cutout specifying member, and an opening pressing member that can be pressed on the surface side of the tissue cutout specifying member so as to encapsulate the cutout specifying opening,
The tissue piece cryopreservation plate has a width that is the same as or slightly wider than the predetermined width of the cutout specifying opening, and extends a predetermined length and mounts the cut tissue piece cut using the collected tissue cutout position specifying tool. A harvested tissue piece vitrified cryopreservation tool characterized in that it is a metal plate-like member that can be placed and has a number of holes through which a cooling medium can enter.

The harvested tissue piece vitrification cryopreservation tool according to claim 1, wherein the harvested tissue piece vitrification cryopreservation tool is an ovarian tissue vitrification cryopreservation tool.

The harvested tissue cutting position specifying tool, after pressing the tissue cutting portion specifying member against the target biological tissue, making a cut along the inner edge of the opening for specifying the cutting portion, then the tissue cutting portion specifying member The opening pressing member is pressed against the surface side of the cutting portion so as to enclose the opening for specifying the cutting portion, and the cut tissue portion is pressed along the back surface of the tissue cutting portion specifying member. The harvested tissue piece vitrified cryopreservation tool according to claim 1 or 2, wherein a tissue piece having a shape corresponding to the shape of the opening can be cut by moving the cutting tool.

The cutout specifying opening has a rectangular shape, and the tissue piece cryopreservation plate has a width the same as or slightly wider than the rectangular opening, and the rectangular opening. The harvested tissue piece vitrified cryopreservation device according to any one of claims 1 to 3, which has a length sufficiently longer than the length of the portion.

The cutout specifying opening is circular or elliptical, and the tissue piece cryopreservation plate has a width the same as or slightly wider than the diameter or short axis of the opening, and the diameter of the opening The collected tissue piece vitrified cryopreservation device according to any one of claims 1 to 3, which has a length sufficiently longer than the length of the major axis.

The harvested tissue piece vitrified cryopreservation according to any one of claims 1 to 5, wherein the locking portion for a cutting tool is provided on an end side of the tissue cut portion specifying member from the cut portion specifying opening. Tools.

The harvested tissue piece vitrified cryopreservation device according to any one of claims 1 to 6, wherein the cutting tool locking portion is provided in the vicinity of the cutout specifying opening.

The harvested tissue piece vitrified cryopreservation tool according to any one of claims 1 to 7, wherein the opening pressing member is a transparent flat plate member.

The harvested tissue piece vitrification according to any one of claims 1 to 8, wherein the tissue piece cryopreservation plate is a substantially rectangular thin plate plate, and has a large number of holes through which a cooling medium can enter. Cryopreservation tool.

The sampling according to any one of claims 1 to 9, wherein the tissue piece cryopreservation plate is formed of a metal selected from copper, copper alloy, aluminum, aluminum alloy, gold, gold alloy, silver, and silver alloy. Tissue piece vitrification cryopreservation tool.

The tissue cutout specifying member is provided on both sides of the surface so as to extend from one end to the other end, and includes an opening pressing member guiding portion that slidably accommodates both sides of the opening pressing member. The harvested tissue piece vitrified cryopreservation device according to any one of claims 1 to 10.

The tissue cutout specifying member is provided on at least one side of the back surface so as to extend from one end side to the other end side, and further includes a cutting tool guide section on which the cutting tool can slide. The collected tissue piece vitrified cryopreservation device according to any one of the above.

The harvested tissue piece vitrification cryopreservation tool according to claim 12, wherein a distal end portion of the cutting tool guiding portion protrudes from one end of the tissue cutting portion specifying member.

It is a metallic plate-like member extending for a predetermined length, and has a tissue piece placement section, and at least the tissue piece placement section has a number of holes through which a cooling medium can enter, for tissue piece cryopreservation plate.

The tissue piece cryopreservation plate according to claim 14, wherein the tissue piece cryopreservation plate is formed of a metal selected from copper, copper alloy, aluminum, aluminum alloy, gold, gold alloy, silver, and silver alloy. .

The tissue piece cryopreservation plate according to claim 14 or 15, wherein the tissue piece cryopreservation plate is a thin plate having a substantially rectangular shape, and has a large number of holes through which a cooling medium can enter.

A harvested tissue piece vitrification cryopreservation method using the harvested tissue piece vitrification cryopreservation device according to any one of claims 1 to 13,
After making a cut along the inner edge of the opening for specifying the cut-out portion in a state where the tissue cut-out specifying member is pressed against the target living tissue, the opening pressing member on the surface side of the tissue cut-out specifying member The opening is moved by moving the cutting tool along the back surface of the tissue cutout specifying member in a state where the cut tissue specifying portion is pressed against the cutout specifying opening. A tissue piece collecting step of cutting out a tissue piece having a shape corresponding to the shape of the part, a tissue piece vitrifying solution immersing step of immersing the collected tissue piece in a vitrification solution, and taking out the tissue piece from the vitrification solution, the tissue Tissue piece placing step for placing on a piece cryopreservation plate, and an input step for putting the tissue piece cryopreservation plate on which the tissue piece is placed into a cooling medium tank Single glass freezing Exist method.

The harvested tissue piece vitrification cryopreservation method according to claim 17, wherein the harvested tissue piece vitrification cryopreservation method is an ovarian tissue vitrification cryopreservation method or a liver tissue vitrification cryopreservation method.