JPH058676B2 - - Google Patents
Info
- Publication number
- JPH058676B2 JPH058676B2 JP62263507A JP26350787A JPH058676B2 JP H058676 B2 JPH058676 B2 JP H058676B2 JP 62263507 A JP62263507 A JP 62263507A JP 26350787 A JP26350787 A JP 26350787A JP H058676 B2 JPH058676 B2 JP H058676B2
- Authority
- JP
- Japan
- Prior art keywords
- inner cylinder
- cells
- container
- cryopreservation
- cylinder
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 238000005138 cryopreservation Methods 0.000 claims description 29
- 210000004102 animal cell Anatomy 0.000 claims description 16
- OVARTBFNCCXQKS-UHFFFAOYSA-N propan-2-one;hydrate Chemical compound O.CC(C)=O OVARTBFNCCXQKS-UHFFFAOYSA-N 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 239000011796 hollow space material Substances 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 description 21
- 238000007710 freezing Methods 0.000 description 9
- 230000008014 freezing Effects 0.000 description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 238000001816 cooling Methods 0.000 description 6
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 239000004698 Polyethylene Substances 0.000 description 2
- 229910052782 aluminium Inorganic materials 0.000 description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 2
- 239000003708 ampul Substances 0.000 description 2
- 244000309466 calf Species 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 239000001569 carbon dioxide Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- -1 polyethylene Polymers 0.000 description 2
- 229920000573 polyethylene Polymers 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241000309219 Sium medium Species 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 239000012595 freezing medium Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 210000004408 hybridoma Anatomy 0.000 description 1
- 239000005457 ice water Substances 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000009877 rendering Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 239000002893 slag Substances 0.000 description 1
- 238000010583 slow cooling Methods 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Description
［産業上の利用分野］
本発明は、動物細胞の凍結保存容器に関するも
のである。
［従来の技術］
最近、動物細胞を利用して、種々の物質を生産
したり、病気の治療薬を開発する事が盛んに行な
われているため、利用する細胞の保存が必要とな
つている。ところで、細胞の寿命は極めて短いた
め、培地で培養しながら、細胞を継代維持しなけ
ればならない。
しかしながら、培養条件は細胞の種類によつて
相違するため、培養には非常に手間がかかり、ま
た、細胞相互に置換や侵入が生じたり、細胞が微
生物に汚染されたりする恐れがある。また継代有
限性の細胞は若い継代歴の時期に保存する必要が
あるし、特に、物質生産を目的とする遺伝子組換
え型の動物細胞や抗体等を産出するハイブリドー
マなどは培養中に性質変化を来すこともあり、貴
重な資源を損なうだけでなく性質変化の有無をチ
エツクしなくては使用できないという問題があ
る。
そこで、細胞を凍結用の培地とともにアンプル
やチユーブ中に封入しこれを凍結して保存する方
法が考えられている。この場合の凍結手段として
は、事業所や実験室で広く使用されている−80℃
程度の超低温槽が用いられることもあるが、−100
℃以下の温度で低ければ低いほど良好な結果が得
られるので最近は液体窒素が用いられるようにな
つている。この方法によれば細胞を培養する必要
がないため、細胞の継代変化を来さない等の長所
がある。
［発明が解決しようとする問題点］
しかしながら、細胞凍結には、急速凍結ではな
く緩徐な凍結が必要とされており、アンプル等を
超低温槽や液体窒素中にそのまま入れると冷却速
度が速すぎて細胞の死滅や失活を来すという問題
があつた。このように死滅や失活を来たした細胞
は利用不能になり凍結保存の意味がなくなつてし
まう。
本発明は、従来用いられている超低温槽をその
まま用いて細胞の緩徐な凍結を行える動物細胞凍
結容器を提供することを目的とする。
［問題点を解決するための手段］
本発明は、その目的を達成させるために、次の
ような構成としている。すなわち、本発明に係る
凍結保存容器は、有底の外筒と、この外筒の内部
に互いに離間した状態で配置された有底の内筒と
を有している。内筒と外筒との間の開口部には蓋
を設け、これら内筒と外筒との間の中空部に水で
希釈して20〜40容量％としたアセトン水を封入し
ている。内筒内には凍結保存液とともに動物細胞
を封入した封入容器を収納し、前記内筒の上部開
口面にはこの開口面を密閉する断熱性の蓋を設け
たものとしている。
［実施例］
以下、本発明の凍結保存容器の一実施例を第１
図を参照しながら説明する。この図において１は
本実施例の凍結保存容器である。この凍結保存容
器１は、有底円筒状の外筒２と、この外筒２の内
部に互いに離間した状態で配置されたやはり有底
円筒状の内筒３とを有している。これら外筒２と
内筒３とは、ともに断熱性の高い材料からなつて
おり、本実施例ではポリエチレン製である。
前記内筒３と外筒２との間の開口部には、この
開口部を塞ぐ蓋４が設けられている。この蓋４
は、内筒３・外筒２と一体に形成されているとと
もに、内筒３と外筒２との間の中空部５と外部と
を連通する孔６が対向する位置に２箇所設けられ
ている。前記中空部５には蒸溜水で希釈して20〜
40容量％、好ましくは30容量％としたアセトン水
７が前記蓋４近傍まで封入されている。
前記内筒３内には、凍結保存液８とともに動物
細胞（図示せず）を封入しかつ中空角柱状のアル
ミニウム製のチユーブから形成された封入容器９
が収納されている。前記凍結保存液８としては、
本実施例では、ジメチルスルフオキサイド
（Dimethyl sulfoxide）10容量％、カーフ フエ
ータルボビン セルム（Calf，Fetal Bovine
Serum）15容量％、イーグルズ エム．イー．エ
ム；イー．アールズベース（Eagle′sM.E.M；
Earle′s−base）75容量％（但し、炭酸ガスおよ
び炭酸水素ナトリウムを使用して凍結保存液８の
PH値をPH6.8〜7.0に調整した）の組成のものが用
いられている。そして、この凍結保存液８に対す
る細胞の密度は１×107〜５×106個／ml程度とす
るのが好ましい。また前記封入容器９としては、
本実施例では中空角柱状のアルミニウム製のチユ
ーブを用いたが、これに限らず中空円柱状でも良
く、また、ガラス・メタル・スラグ等でできたア
ンプルでも良い。
前記内筒３の上部開口面３ａには、この開口面
３ａを密閉する断熱性の蓋１０が載置されてい
る。この蓋１０は、外筒２と等しい径の円形板状
に形成されたもので、その下側面１０ａに、前記
内筒３の内径と同じかごくわずかに小さい径の円
柱状の嵌合部１１が設けられている。この嵌合部
１１は前記内筒３の内側に嵌合して内筒３内部を
気密に保つものである。また、このとき蓋１０の
下側面１０ａによつて蓋４の孔６が塞がれること
になる。
つぎに、本実施例の凍結保存容器１の使用方法
を説明する。
まず、保存の対象となる動物細胞を凍結保存液
８に浮遊させ、封入容器９に充填したのち封入容
器９の開口部を熔封する。つぎに封入容器９を内
筒３の底面上に載置して収納してから、蓋１０の
嵌合部１１を内筒３に嵌合させて内筒３の開口面
３ａを密閉する。
ついで、凍結保存容器１を−80℃の超低温槽に
収納する。すると凍結保存容器１内の動物細胞
は、緩徐な冷却速度で冷却され凍結されて、その
温度が−80℃に到達し、この温度で保存される。
なお、さらに低温で保存する必要のあるとき
は、細胞を凍結保存容器１に収納してから約２〜
４時間後に液体窒素タンクに移入する。
また、上記のように凍結された細胞を解凍して
再培養するには、次のようにする。まず超低温槽
中の凍結保存容器１から封入容器９を取り出し、
温水のなかで激しくゆすりながら凍結保存液８を
解凍し、その後直ちに氷水中にいれて冷却する。
ついで、封入容器９中の細胞を取り出した後、遠
心分離機で洗浄し、冷却していない増殖用培養液
を細胞に加え、適当な細胞数に調整した後培養す
る。
前記の冷却速度としては、−１℃／min程度が
望ましいことが実験的に確かめられており、本実
施例の保存容器１では、第２図に示すようにこの
冷却速度にきわめて近い値が得られる。第２図の
データは次のような条件下で得られた。
［条件］
外筒の高さ 72mm
外筒の幅 62mm
内筒の高さ 62mm
内筒の幅 47mm
内筒と外筒の素材 ポリエチレン
内筒と外筒の中空部の容量 100ml
アセトン水の容量 100ml
超低温槽の温度 −80℃（一定）
凍結保存板（エフ．エス．培地）の組成
ジメチルスルフオキサイド10容量％
カーフ フエータル ボビン セルム15容量％
イーグルズ エム．イー．エム（アールズベー
ス）75容量％
PH6.8〜7.0（炭酸ガスおよび炭酸水素ナトリウム
の添加によりPH値を調整した）
つぎに、上記実験例の凍結保存容器１を使用し
て保存した動物細胞の生存率を第１表〜第３表に
示す。
[Industrial Application Field] The present invention relates to a cryopreservation container for animal cells. [Prior art] Recently, animal cells have been actively used to produce various substances and to develop therapeutic drugs for diseases, so it has become necessary to preserve the cells to be used. . By the way, since the lifespan of cells is extremely short, cells must be maintained for passage while being cultured in a medium. However, since the culture conditions differ depending on the type of cells, culturing is extremely time-consuming and there is a risk that cells may be replaced or invaded by each other, or cells may be contaminated with microorganisms. In addition, cells with limited passage need to be preserved at a young passage history, and in particular, genetically modified animal cells for the purpose of producing substances and hybridomas that produce antibodies etc. There is a problem that not only does it damage valuable resources, but also that it cannot be used without checking for changes in properties. Therefore, methods have been considered in which cells are sealed in ampoules or tubes together with a freezing medium and then frozen and preserved. In this case, the freezing method is -80℃, which is widely used in business offices and laboratories.
Although ultra-low temperature chambers with a temperature of -100
Recently, liquid nitrogen has come to be used because the lower the temperature (below ℃), the better the results. According to this method, there is no need to culture the cells, so there are advantages such as no change in cell passage. [Problems to be solved by the invention] However, cell freezing requires slow freezing rather than rapid freezing, and if the ampoule or the like is placed directly into a cryostat or liquid nitrogen, the cooling rate will be too fast. There was a problem with cell death and deactivation. Cells that die or become inactivated in this way become unusable, rendering cryopreservation meaningless. An object of the present invention is to provide an animal cell freezing container that can slowly freeze cells using a conventionally used cryogenic chamber as is. [Means for Solving the Problems] In order to achieve the object, the present invention has the following configuration. That is, the cryopreservation container according to the present invention has an outer cylinder with a bottom and an inner cylinder with a bottom which is arranged inside the outer cylinder so as to be spaced apart from each other. A lid is provided at the opening between the inner cylinder and the outer cylinder, and acetone water diluted with water to a concentration of 20 to 40% by volume is sealed in the hollow space between the inner cylinder and the outer cylinder. The inner cylinder contains an enclosure containing animal cells together with a cryopreservation solution, and the upper opening of the inner cylinder is provided with an insulating lid for sealing the opening. [Example] Hereinafter, a first example of the cryopreservation container of the present invention will be described.
This will be explained with reference to the figures. In this figure, 1 is the cryopreservation container of this example. This cryopreservation container 1 has an outer tube 2 having a cylindrical shape with a bottom, and an inner tube 3 which also has a cylindrical shape and a bottom and is arranged inside the outer tube 2 so as to be spaced apart from each other. Both the outer cylinder 2 and the inner cylinder 3 are made of a material with high heat insulation properties, and in this embodiment, they are made of polyethylene. A lid 4 is provided at the opening between the inner cylinder 3 and the outer cylinder 2 to close this opening. This lid 4
is formed integrally with the inner cylinder 3 and the outer cylinder 2, and has two holes 6 at opposing positions that communicate the hollow part 5 between the inner cylinder 3 and the outer cylinder 2 with the outside. There is. In the hollow part 5, diluted with distilled water, 20~
Acetone water 7 of 40% by volume, preferably 30% by volume is filled up to the vicinity of the lid 4. Inside the inner cylinder 3, there is an enclosure container 9 formed from a hollow prismatic aluminum tube, in which animal cells (not shown) are enclosed together with a cryopreservation solution 8.
is stored. As the cryopreservation solution 8,
In this example, 10% by volume of dimethyl sulfoxide, Calf, Fetal Bovine
Serum) 15% by volume, Eagles M. E. M;E. Eagle′sM.EM;
Earle's-base) 75% by volume (however, using carbon dioxide gas and sodium bicarbonate to prepare cryopreservation solution 8)
A composition with a pH value adjusted to 6.8 to 7.0) is used. The density of cells in this cryopreservation solution 8 is preferably about 1×10 7 to 5×10 6 cells/ml. Further, as the enclosure container 9,
In this embodiment, a hollow prismatic aluminum tube is used, but the tube is not limited to this, and may be a hollow cylinder, or an ampoule made of glass, metal, slag, or the like. A heat insulating lid 10 is placed on the upper opening surface 3a of the inner cylinder 3 to seal the opening surface 3a. This lid 10 is formed into a circular plate shape with a diameter equal to that of the outer cylinder 2, and a cylindrical fitting part 11 with a diameter equal to or slightly smaller than the inner diameter of the inner cylinder 3 is formed on the lower side 10a. is provided. This fitting part 11 fits inside the inner cylinder 3 to keep the inside of the inner cylinder 3 airtight. Further, at this time, the hole 6 of the lid 4 is closed by the lower surface 10a of the lid 10. Next, a method of using the cryopreservation container 1 of this example will be explained. First, animal cells to be preserved are suspended in a cryopreservation solution 8 and filled into an enclosure container 9, and then the opening of the enclosure container 9 is sealed. Next, the enclosure 9 is placed on the bottom surface of the inner tube 3 and stored, and then the fitting part 11 of the lid 10 is fitted into the inner tube 3 to seal the opening surface 3a of the inner tube 3. Then, the cryopreservation container 1 is stored in an ultra-low temperature chamber at -80°C. Then, the animal cells in the cryopreservation container 1 are cooled and frozen at a slow cooling rate until the temperature reaches -80°C and is stored at this temperature. If it is necessary to preserve the cells at an even lower temperature, store the cells in the cryopreservation container 1 for about 2 to 30 minutes.
Transfer to liquid nitrogen tank after 4 hours. Furthermore, to thaw and re-culture the frozen cells as described above, proceed as follows. First, take out the enclosure container 9 from the cryopreservation container 1 in the cryogenic chamber,
Thaw the cryopreservation solution 8 by shaking vigorously in warm water, and then immediately cool it in ice water.
Next, after the cells in the enclosure container 9 are taken out, they are washed with a centrifuge, and an uncooled growth culture medium is added to the cells to adjust the cell number to an appropriate number, followed by culturing. It has been experimentally confirmed that the above-mentioned cooling rate is desirably about -1°C/min, and the storage container 1 of this example has a cooling rate that is very close to this cooling rate, as shown in Fig. 2. It will be done. The data in Figure 2 were obtained under the following conditions. [Conditions] Height of outer cylinder 72mm Width of outer cylinder 62mm Height of inner cylinder 62mm Width of inner cylinder 47mm Material of inner cylinder and outer cylinder Polyethylene Capacity of hollow part of inner cylinder and outer cylinder 100ml Capacity of acetone water 100ml Ultra-low temperature Temperature of tank -80℃ (constant) Composition of cryopreservation plate (F.S. medium) Dimethyl sulfoxide 10% by volume Calf Fatal bobbin Cellum 15% by volume Eagles M. E. M (R's Base) 75% by volume PH6.8-7.0 (PH value was adjusted by adding carbon dioxide gas and sodium bicarbonate) Next, the survival of animal cells preserved using cryopreservation container 1 of the above experimental example The rates are shown in Tables 1 to 3.
【表】【table】
【表】【table】
【表】
以上説明したように、本実施例の凍結保存容器
１によれば、従来から事業所や研究室等で広く用
いられている超低温槽を用いて、動物細胞の緩徐
な凍結を好ましい冷却速度下において行うことが
できるという利点がある。特に本実施例の凍結保
存容器１によれば、なんら機械的あるいは電気的
な制御手段を用いずに簡易な手段で好ましい冷却
速度を得ることができる。
［発明の効果］
本発明は、それぞれ有底の外筒と内筒との間の
開口部に蓋を設け、これら内筒と外筒との間の中
空部に水で希釈して20〜40容量％としたアセトン
水を封入し、内筒内に動物細胞を封入した封入容
器を収納し、前記内筒の上部開口面に断熱性の蓋
を設けた動物細胞の冷凍保存容器に構成されるの
で、簡単な構造でありながら超底温槽に容器を収
納するだけて動物細胞の理想的に緩徐な凍結を行
うことができ、凍結後の動物細胞の生存率を高め
ることができるだけでなく、加えて凍結年数や移
植性等にも好ましい結果が得られるというすぐれ
た効果を奏する。[Table] As explained above, according to the cryopreservation container 1 of the present example, slow freezing of animal cells can be achieved using a cryogenic chamber that has been widely used in business offices and laboratories. It has the advantage that it can be carried out at low speeds. In particular, according to the cryopreservation container 1 of this embodiment, a preferable cooling rate can be obtained by simple means without using any mechanical or electrical control means. [Effects of the Invention] The present invention provides a cover at the opening between the bottomed outer cylinder and the inner cylinder, and fills the hollow part between the inner cylinder and the outer cylinder with water diluted with A cryopreservation container for animal cells is constructed by enclosing acetone water (volume %), storing an enclosure containing animal cells in an inner cylinder, and providing an insulating lid on the upper opening surface of the inner cylinder. Therefore, although it has a simple structure, it is possible to perform ideal slow freezing of animal cells by simply storing the container in an ultrathermal bath, and it not only increases the survival rate of animal cells after freezing. In addition, it has excellent effects in that favorable results can be obtained in terms of freezing years, portability, etc.
第１図は、本発明の凍結保存容器の一実施例を
示す断面図、第２図は本実施例の凍結保存容器を
使用したときの冷却速度を示すグラフである。
１……凍結保存容器、２……外筒、３……内
筒、４……開口部に設けられた蓋、５……中空
部、７……アセトン水、８……凍結保存液、９…
…封入容器、１０……開口面を密閉する蓋。
FIG. 1 is a cross-sectional view showing one embodiment of the cryopreservation container of the present invention, and FIG. 2 is a graph showing the cooling rate when the cryopreservation container of this example is used. 1... Cryopreservation container, 2... Outer tube, 3... Inner tube, 4... Lid provided at the opening, 5... Hollow part, 7... Acetone water, 8... Cryopreservation solution, 9 …
...Enclosed container, 10...Lid that seals the opening surface.
Claims (1)
した状態で配置された有底の内筒とを有し、内筒
と外筒との間の開口部に蓋を設け、これら内筒と
外筒との間の中空部に水で希釈して20〜40容量％
としたアセトン水を封入し、内筒内に凍結保存液
とともに動物細胞を封入した封入容器を収納し、
前記内筒の上部開口面にこの開口面を密閉する断
熱性の蓋を設けたことを特徴とする動物細胞の凍
結保存容器。1. It has an outer cylinder with a bottom and an inner cylinder with a bottom which is arranged inside the outer cylinder in a spaced manner, and a cover is provided at the opening between the inner cylinder and the outer cylinder, and these inner cylinders are provided with a lid. Dilute with water to 20 to 40% by volume in the hollow space between the cylinder and the outer cylinder.
A container containing acetone water and animal cells sealed together with a cryopreservation solution is placed in the inner cylinder.
A cryopreservation container for animal cells, characterized in that an insulating lid is provided on the upper opening surface of the inner cylinder to seal the opening surface.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62263507A JPH01104163A (en) | 1987-10-19 | 1987-10-19 | Container for freezing and storing animal cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62263507A JPH01104163A (en) | 1987-10-19 | 1987-10-19 | Container for freezing and storing animal cell |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH01104163A JPH01104163A (en) | 1989-04-21 |
| JPH058676B2 true JPH058676B2 (en) | 1993-02-02 |
Family
ID=17390488
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP62263507A Granted JPH01104163A (en) | 1987-10-19 | 1987-10-19 | Container for freezing and storing animal cell |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH01104163A (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006059626A1 (en) * | 2004-12-03 | 2006-06-08 | Nipro Corporation | Device for biosample storage |
| JP5607896B2 (en) * | 2009-05-12 | 2014-10-15 | 株式会社日立製作所 | Cryopreservation method, freezing device, cryopreservation system |
| US12038133B2 (en) | 2020-07-23 | 2024-07-16 | Coopersurgical, Inc. | Canister caps for cryopreservation applications |
-
1987
- 1987-10-19 JP JP62263507A patent/JPH01104163A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPH01104163A (en) | 1989-04-21 |
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