JPH01229046A - Production of gel - Google Patents
Production of gelInfo
- Publication number
- JPH01229046A JPH01229046A JP5383288A JP5383288A JPH01229046A JP H01229046 A JPH01229046 A JP H01229046A JP 5383288 A JP5383288 A JP 5383288A JP 5383288 A JP5383288 A JP 5383288A JP H01229046 A JPH01229046 A JP H01229046A
- Authority
- JP
- Japan
- Prior art keywords
- gel
- solution
- cells
- sodium alginate
- animal cells
- 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.)
- Granted
Links
- 238000004519 manufacturing process Methods 0.000 title claims description 5
- 239000000499 gel Substances 0.000 claims abstract description 31
- 210000004027 cell Anatomy 0.000 claims abstract description 21
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims abstract description 17
- 239000000661 sodium alginate Substances 0.000 claims abstract description 17
- 235000010413 sodium alginate Nutrition 0.000 claims abstract description 17
- 229940005550 sodium alginate Drugs 0.000 claims abstract description 17
- 210000004102 animal cell Anatomy 0.000 claims abstract description 15
- 229910001427 strontium ion Inorganic materials 0.000 claims abstract description 9
- 239000010419 fine particle Substances 0.000 claims abstract description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 5
- 229920002472 Starch Polymers 0.000 claims abstract description 5
- 239000011324 bead Substances 0.000 claims abstract description 5
- 239000011521 glass Substances 0.000 claims abstract description 5
- 239000008107 starch Substances 0.000 claims abstract description 5
- 235000019698 starch Nutrition 0.000 claims abstract description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 3
- 239000003456 ion exchange resin Substances 0.000 claims abstract description 3
- 229920003303 ion-exchange polymer Polymers 0.000 claims abstract description 3
- 239000000741 silica gel Substances 0.000 claims abstract description 3
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 3
- 210000002257 embryonic structure Anatomy 0.000 claims description 4
- 230000000392 somatic effect Effects 0.000 claims description 4
- 210000005061 intracellular organelle Anatomy 0.000 claims description 3
- 239000000126 substance Substances 0.000 abstract description 6
- 238000000465 moulding Methods 0.000 abstract description 3
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 abstract description 2
- 239000000203 mixture Substances 0.000 abstract description 2
- PWYYWQHXAPXYMF-UHFFFAOYSA-N strontium(2+) Chemical compound [Sr+2] PWYYWQHXAPXYMF-UHFFFAOYSA-N 0.000 abstract 2
- 229910052799 carbon Inorganic materials 0.000 abstract 1
- 210000001161 mammalian embryo Anatomy 0.000 abstract 1
- 238000002156 mixing Methods 0.000 abstract 1
- 210000000056 organ Anatomy 0.000 abstract 1
- 235000013311 vegetables Nutrition 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 16
- 239000013543 active substance Substances 0.000 description 5
- 241000699800 Cricetinae Species 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- AEMOLEFTQBMNLQ-UHFFFAOYSA-N beta-D-galactopyranuronic acid Natural products OC1OC(C(O)=O)C(O)C(O)C1O AEMOLEFTQBMNLQ-UHFFFAOYSA-N 0.000 description 4
- 210000003292 kidney cell Anatomy 0.000 description 4
- 229910001631 strontium chloride Inorganic materials 0.000 description 4
- AHBGXTDRMVNFER-UHFFFAOYSA-L strontium dichloride Chemical compound [Cl-].[Cl-].[Sr+2] AHBGXTDRMVNFER-UHFFFAOYSA-L 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- 235000010443 alginic acid Nutrition 0.000 description 3
- 229920000615 alginic acid Polymers 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 230000000704 physical effect Effects 0.000 description 3
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 2
- AEMOLEFTQBMNLQ-BZINKQHNSA-N D-Guluronic Acid Chemical compound OC1O[C@H](C(O)=O)[C@H](O)[C@@H](O)[C@H]1O AEMOLEFTQBMNLQ-BZINKQHNSA-N 0.000 description 2
- 206010020649 Hyperkeratosis Diseases 0.000 description 2
- 229940072056 alginate Drugs 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- AEMOLEFTQBMNLQ-YBSDWZGDSA-N d-mannuronic acid Chemical compound O[C@@H]1O[C@@H](C(O)=O)[C@H](O)[C@@H](O)[C@H]1O AEMOLEFTQBMNLQ-YBSDWZGDSA-N 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000003349 gelling agent Substances 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 210000001938 protoplast Anatomy 0.000 description 2
- 229910052712 strontium Inorganic materials 0.000 description 2
- CIOAGBVUUVVLOB-UHFFFAOYSA-N strontium atom Chemical compound [Sr] CIOAGBVUUVVLOB-UHFFFAOYSA-N 0.000 description 2
- 229960001126 alginic acid Drugs 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 238000005266 casting Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 210000004408 hybridoma Anatomy 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 210000001589 microsome Anatomy 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- QPILZZVXGUNELN-UHFFFAOYSA-N sodium;4-amino-5-hydroxynaphthalene-2,7-disulfonic acid Chemical compound [Na+].OS(=O)(=O)C1=CC(O)=C2C(N)=CC(S(O)(=O)=O)=CC2=C1 QPILZZVXGUNELN-UHFFFAOYSA-N 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Manufacture Of Porous Articles, And Recovery And Treatment Of Waste Products (AREA)
- Compositions Of Macromolecular Compounds (AREA)
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は、アルギン酸ストロンチウムのゲルを製造する
方法に関するものである。DETAILED DESCRIPTION OF THE INVENTION (Industrial Field of Application) The present invention relates to a method for producing a strontium alginate gel.
更に詳細には、本発明は、ゲル内に多数の空ゲキを有す
るアルギン酸ストロンチウムのゲルを製造する方法に関
するものである。More particularly, the present invention relates to a method for producing a strontium alginate gel having a large number of voids within the gel.
本発明によって製造されたゲルはゲル内に多数の空ゲキ
を有し、ここに動物細胞等の生理活性体を添加した場合
には、生理活性体が大量に増殖することができるために
、動物細胞等の生理活性体が生産する物質を大量に生産
させることが容易となり、発酵工業界、製薬界等に寄与
するところ大なるものがある。The gel produced according to the present invention has a large number of empty cells within the gel, and when bioactive substances such as animal cells are added thereto, the bioactive substances can proliferate in large quantities. It has become easier to mass-produce substances produced by physiologically active substances such as cells, and this will greatly contribute to the fermentation industry, pharmaceutical industry, etc.
（従来技術）
一般に動物細胞、植物細胞その他これらに関連した生理
活性体などを固定化するには、これらの１種もしくは２
種以上をアルギン酸ナトリウムなどのゲル化能を有する
物質の水溶液に懸濁し、Ｃａ”＋などのゲル化剤中に滴
下してゲル化して、固定化が行なわれている。(Prior art) Generally, in order to immobilize animal cells, plant cells, and other physiologically active substances related to these, one or two of these are used.
Immobilization is carried out by suspending a species or more in an aqueous solution of a substance with gelling ability such as sodium alginate, and dropping the suspension into a gelling agent such as Ca''+ to form a gel.
（発明が解決しようとする問題点）
従来技術によって、動物細胞等を固定化する場合、自由
に増殖するだけ多数の空ゲキが与えられることはなかっ
た。適当な空ゲキがないために一定の許容範囲まで増殖
すると、動物細胞等は増殖を停止してしまって、培養生
産物を生産することもなくなってしまうのであった。(Problems to be Solved by the Invention) When animal cells and the like are immobilized using conventional techniques, they are not provided with enough cells to proliferate freely. Due to the lack of adequate energy, animal cells, etc., once they proliferate to a certain permissible range, stop proliferating and no longer produce any cultured products.
（問題点を解決するための手段）
本発明者らは、ゲルの中に多数の空ゲキを作ることがで
きれば、動物細胞等を固定化して培養する際、著しるし
く物質生産性を高めることができるとの考えから鋭意研
究したところ、特定の物性をそなえた゛アルギン酸ナト
リウムを使用すれば、固定化したとき自然に多数の空ゲ
キを形成させることができることを知ったのである。(Means for Solving the Problems) The present inventors believe that if a large number of empty cells can be created in a gel, it will significantly increase material productivity when immobilizing and culturing animal cells, etc. After conducting intensive research with the idea that this could be possible, he discovered that if sodium alginate, which has specific physical properties, was used, it would be possible to naturally form a large number of air bubbles when immobilized.
本発明は、この知見から完成されたもので、Ｍ／Ｇ比が
０．１〜０．５で、１％溶液で１０〜１５００ｃｐ（２
５℃における）であるアルギン酸ナトリウムの０．１〜
２％の溶液に、動物細胞、細胞内小器官、植物細胞。The present invention was completed based on this knowledge, and has an M/G ratio of 0.1 to 0.5 and a 1% solution of 10 to 1500 cp (2
0.1~ of sodium alginate) at 5°C
Animal cells, organelles, and plant cells in a 2% solution.
植物の不定胚、デンプン、ガラスビーズ、活性炭、シリ
カゲル、イオン交換樹脂などの微粒子の１種以上を添加
したものをストロンチウムイオンで適宜形状に成型する
ことを特徴とする空ゲキを有するゲルの製法である。A method for producing a gel with air bubbles, which is characterized by molding a gel containing one or more of fine particles such as plant somatic embryos, starch, glass beads, activated carbon, silica gel, and ion exchange resin into an appropriate shape using strontium ions. be.
微粒子の大きさとしては０．１〜２００μｍ程度がよし
為。The size of the fine particles is preferably about 0.1 to 200 μm.
本発明の空ゲキとは、ゲル中に微粒子が点在しており、
その微粒子からゲルの中心に向って空ゲキができている
ものであり、空ゲキの中は、水や培地などの液体が充満
しているものである。The dry gel of the present invention has fine particles scattered in the gel,
An air bubble is formed from the fine particles toward the center of the gel, and the air bubble is filled with liquid such as water or a culture medium.
本発明においては、特定の物性をそなえたアルギン酸ナ
トリウムが使用される。In the present invention, sodium alginate with specific physical properties is used.
一般に、アルギン酸はＤ−マンヌロン酸（Ｍ）とし−グ
ルロン酸（Ｇ）とから構成されていて、両者の成分比（
Ｍ／Ｇ比）が物性に大きな影響を与えている。Generally, alginic acid is composed of D-mannuronic acid (M) and guluronic acid (G), and the ratio of the two components is (
M/G ratio) has a great influence on physical properties.
そして、Ｍのみからなる部分、Ｇのみからなる部分、Ｍ
とＧが混在する部分などがあるがゲル化剤の金属イオン
と結合して強固なゲルを形成するのは主としてＧブロッ
クの働きによることが明らかとなっている。Then, a part consisting only of M, a part consisting only of G, M
Although there are parts where G and G are mixed, it is clear that it is mainly the function of the G block that forms a strong gel by combining with the metal ions of the gelling agent.
本発明では、アルギン酸を構成するＤ−マンヌロン酸（
Ｍ）とし−グルロン酸（Ｇ）の比、すなわちＭ／Ｇ比が
０．１〜０．５、好ましくは０．１〜０．４のものを選
んで用いる。In the present invention, D-mannuronic acid (
M) Those having a ratio of ash and guluronic acid (G), that is, an M/G ratio of 0.1 to 0.5, preferably 0.1 to 0.4, are selected and used.
また、本発明に用いるアルギン酸ナトリウムは１％溶液
で、２５℃で測定した粘度が１０〜１５００ｃｐ、好ま
しくは５０〜５００ｃｐのものでなければならない。Furthermore, the sodium alginate used in the present invention is a 1% solution and must have a viscosity of 10 to 1500 cp, preferably 50 to 500 cp, as measured at 25°C.
また、成型時のアルギン酸ナトリウムの溶液の濃度は０
．１〜２％、好ましくは０．５〜１．０％でなければな
らない。Also, the concentration of sodium alginate solution during molding is 0.
．． It should be between 1 and 2%, preferably between 0.5 and 1.0%.
以上のＭ／Ｇ比、粘度及び溶液の濃度すべてが上記特定
の範囲にあるとき、動物細胞等を添加混合して、ストロ
ンチウムイオンによって成型して、はじめてゲル内に多
数の空ゲキを形成させることができる。多数の空ゲキを
形成させるための条件は多くの実験を重ねて確認された
もので、これらの条件をはずれれば多数の空ゲキは形成
されないことも実験によって確認されたのである。When the above M/G ratio, viscosity, and concentration of the solution are all within the above specific ranges, animal cells, etc. are added and mixed, and formed by strontium ions to form a large number of empty cells in the gel. Can be done. The conditions for forming a large number of air bubbles have been confirmed through many experiments, and it has also been confirmed through experiments that if these conditions are violated, large numbers of air bubbles will not form.
固定化する生理活性体としては、動物細胞、細胞内の小
器官、植物細胞、植物の不定胚がある。Physiologically active substances to be immobilized include animal cells, intracellular organelles, plant cells, and somatic embryos of plants.
動物細胞としては、各種生理活性物質を生産する細胞株
、抗体を生産するハイブリドーマなどがあり、単細胞で
もよく、細胞集合体でもよい、細胞内小器官としてはミ
トコンドリア、ミクロソームなどがある。また、植物細
胞としては各種物質を生産する細胞やカルスがあり、植
物の不定胚としてはカルス細胞、プロトプラスト、巧、
花粉、珠心、子房、胚珠などから誘導したものがある。Animal cells include cell lines that produce various physiologically active substances, hybridomas that produce antibodies, etc., and may be single cells or cell aggregates, and intracellular organelles include mitochondria, microsomes, etc. In addition, plant cells include cells that produce various substances and callus, and somatic embryos of plants include callus cells, protoplasts, protoplasts,
There are those derived from pollen, nucule, ovary, ovule, etc.
動物細胞等はアルギン酸ナトリウム溶液に添加され、混
合してストロンチウムイオンによって適宜形状に成型さ
れる。使用する液としては０．０２５〜１．０モル程度
の塩化ストロンチウム溶液などがよい。Animal cells and the like are added to a sodium alginate solution, mixed, and shaped into an appropriate shape using strontium ions. The liquid to be used is preferably a 0.025 to 1.0 mol strontium chloride solution.
１、 注射器やピペット等のノズルからストロンチウム
イオンを含有する水溶液中に滴下することによりビーズ
状のゲルが得られる。1. A bead-shaped gel can be obtained by dropping it into an aqueous solution containing strontium ions from a nozzle such as a syringe or pipette.
２、 ストロンチウムイオンを含有する水溶液中で注射
器、ピペット等のノズルから連続的に吐出させることに
より繊維状のゲルが得られる。2. A fibrous gel can be obtained by continuously discharging an aqueous solution containing strontium ions from a nozzle of a syringe, pipette, etc.
３、平板上にキャストするか濾紙もしくはガーゼなどに
含浸させた後、水溶液中のストロンチウムイオンと接触
させることにより膜状のゲルを得ることができる。3. After casting on a flat plate or impregnating filter paper or gauze, a membrane-like gel can be obtained by contacting with strontium ions in an aqueous solution.
ここに得られた固定化ゲルはゲル内に多数の空ゲキを有
しており、動物細胞等をかなり長期間にわたって自由に
増殖させることができ、各種生理活性物質等を大量生産
することができるものである。The immobilized gel obtained here has a large number of air cells within the gel, allowing animal cells to grow freely over a fairly long period of time, and making it possible to mass-produce various physiologically active substances. It is something.
次に本発明の実施例を示す。Next, examples of the present invention will be shown.
実施例１
アルギン酸ナトリウム（Ｍ／Ｇ比０．２．１％粘度５０
ｃｐ／２５℃）溶液１００ｍＱに市販の幼ハムスター腎
細胞（大日本製薬に、Ｋ１１）を４．７Ｘ１０’ケ／ｇ
ゲルになるよう添加、混合し最終的に幼ハムスター腎細
胞を懸濁させた１％アルギン酸ナトリウム溶液を得た。Example 1 Sodium alginate (M/G ratio 0.2.1% viscosity 50
Commercially available young hamster kidney cells (Dainippon Pharmaceutical Co., Ltd., K11) were added at 4.7 x 10' cells/g in 100 mQ of solution (cp/25°C).
The mixture was added and mixed to form a gel, and finally a 1% sodium alginate solution in which young hamster kidney cells were suspended was obtained.
この１％アルギン酸ナトリウム溶液を注射器につめ、氷
水中の０．１Ｍ塩化ストロンチウム溶液に滴下し、６０
分放置し、空ゲキが多数できた粒状化ゲルを得た。Fill a syringe with this 1% sodium alginate solution, drop it into a 0.1M strontium chloride solution in ice water, and
After standing for a minute, a granulated gel with many empty particles was obtained.
得られたゲルをカラムにつめ、ＢＭＥ培地３００＋ａＱ
づつを毎日とりかえながら、３０℃で循環培養したとこ
ろ、はとんど変化なく２０日間８０％のグルコースを消
費しているのが認められた。The obtained gel was packed in a column and BME medium 300+aQ
When the cells were cultured in circulation at 30°C while changing the amount of glucose every day, it was observed that the cells consumed 80% of the glucose for 20 days with almost no change.
実施例２
粒径８４μ論を有するデンプン粒０．１％Ｗ／Ｖを含む
１．０％アルギン酸ナトリウム（Ｍ／Ｇ比０．２．１％
粘度５００ｃｐ／２５℃）溶液５０ｔＱを０．１Ｍ塩化
ストロンチウム溶液に滴下し、ビーズ状のゲルを得た。Example 2 1.0% sodium alginate (M/G ratio 0.2.1%) containing 0.1% W/V starch granules with particle size 84 μm
50tQ of the solution (viscosity 500cp/25°C) was dropped into a 0.1M strontium chloride solution to obtain a bead-shaped gel.
得られたゲル内には、デンプン粒から生成した針状の空
ゲキが中心部に向って多数みられた。In the resulting gel, many needle-like cavities generated from starch granules were observed toward the center.
実施例３
アルギン酸ナトリウム（Ｍ／Ｇ比０．２．１％粘度５０
ｃｐ／２５℃）溶液１００ｍＱに市販の幼ハムスター腎
細胞（大日本製薬に、に製）を４．７ Ｘ １０’ケ／
ｇゲルになるよう添加し、更に平均粒径８０μｍを有す
るガラスビーズを０．１％Ｗ／Ｖになるよう添加、両者
を混合し、最終的に幼ハムスター腎細胞及びガラスビー
ズを懸濁させた１％アルギン酸ナトリウム溶液を得た。Example 3 Sodium alginate (M/G ratio 0.2.1% viscosity 50
Commercially available young hamster kidney cells (manufactured by Dainippon Pharmaceutical Co., Ltd.) were added to 4.7
Then, glass beads with an average particle size of 80 μm were added to give a concentration of 0.1% W/V, and both were mixed to finally suspend the young hamster kidney cells and glass beads. A 1% sodium alginate solution was obtained.
この１％アルギン酸ナトリウム溶液を注射器につめ、氷
水中の０．１Ｍ塩化ストロンチウム溶液に滴下し、６０
分放置し、大きな空ゲキを多数有する粒状化ゲルを得た
。Fill a syringe with this 1% sodium alginate solution, drop it into a 0.1M strontium chloride solution in ice water, and
A granulated gel having many large voids was obtained.
得られたゲルをカラムにつめ、ＤＭＥ培地３００社づつ
を毎日とりかえながら、３０℃で循環培養したところ、
はとんど変化なく２０日間８０％のグルコースを消費し
ているのが認められた。The obtained gel was packed in a column and cultured in a circulation at 30°C while replacing 300 DME medium every day.
It was observed that 80% of the glucose was consumed for 20 days with little change.
Claims (1)
１５００ｃｐ（２５℃における）であるアルギン酸ナト
リウムの０．１〜２％の溶液に、動物細胞、細胞内小器
官、植物細胞、植物の不定胚、デンプン、ガラスビーズ
、活性炭、シリカゲル、イオン交換樹脂などの微粒子の
１種以上を添加したものをストロンチウムイオンで適宜
形状に成型することを特徴とする空ゲキを有するゲルの
製法。(1) M/G ratio is 0.1~0.5, 1% solution is 10~
Animal cells, intracellular organelles, plant cells, plant somatic embryos, starch, glass beads, activated carbon, silica gel, ion exchange resins, etc. are added to a 0.1-2% solution of sodium alginate at 1500 cp (at 25°C). 1. A method for producing a gel having air bubbles, which is characterized in that a gel containing one or more types of fine particles is molded into an appropriate shape using strontium ions.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP5383288A JPH01229046A (en) | 1988-03-09 | 1988-03-09 | Production of gel |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP5383288A JPH01229046A (en) | 1988-03-09 | 1988-03-09 | Production of gel |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH01229046A true JPH01229046A (en) | 1989-09-12 |
| JPH0571618B2 JPH0571618B2 (en) | 1993-10-07 |
Family
ID=12953762
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP5383288A Granted JPH01229046A (en) | 1988-03-09 | 1988-03-09 | Production of gel |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH01229046A (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59206041A (en) * | 1983-04-15 | 1984-11-21 | デイモン・バイオテク・インコ−ポレイテツド | Lasting release system |
-
1988
- 1988-03-09 JP JP5383288A patent/JPH01229046A/en active Granted
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59206041A (en) * | 1983-04-15 | 1984-11-21 | デイモン・バイオテク・インコ−ポレイテツド | Lasting release system |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0571618B2 (en) | 1993-10-07 |
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| Date | Code | Title | Description |
|---|---|---|---|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| LAPS | Cancellation because of no payment of annual fees |