JPH0646840A - Solution for freezing and storing cell - Google Patents
Solution for freezing and storing cellInfo
- Publication number
- JPH0646840A JPH0646840A JP4228001A JP22800192A JPH0646840A JP H0646840 A JPH0646840 A JP H0646840A JP 4228001 A JP4228001 A JP 4228001A JP 22800192 A JP22800192 A JP 22800192A JP H0646840 A JPH0646840 A JP H0646840A
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- Prior art keywords
- polymer compound
- solution
- cells
- cell
- cell cryopreservation
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Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は培養動物細胞を簡易な操
作で長期間凍結保存可能な細胞凍結保存液に関する。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a cell cryopreservation liquid capable of cryopreserving cultured animal cells for a long period of time by a simple operation.
【０００２】[0002]
【従来の技術】培養細胞凍結保存は、継代による細胞変
質の防止や雑菌による汚染の防止並びに継代維持の繁雑
さの解消のために行なわれている。これまでの凍結保存
技術は、５〜１５％ＤＭＳＯやグリセリンを含む培養液
に細胞を懸濁しクライオチューブやアンプルにつめ、プ
ログラムフリーザーを用い−１℃／ｍｉｎ前後の速度で
冷却し、最終的に液体窒素中（−１９６℃）での保存方
法が一般的である。2. Description of the Related Art Cryopreservation of cultured cells is carried out in order to prevent alteration of cells due to passage, prevention of contamination by miscellaneous bacteria, and elimination of complexity of passage maintenance. The conventional cryopreservation technology has been to suspend cells in a culture medium containing 5 to 15% DMSO and glycerin, pack them in a cryotube or ampoule, cool them at a rate of around -1 ° C / min using a program freezer, and finally The method of storage in liquid nitrogen (-196 ° C) is common.
【０００３】[0003]
【発明が解決しようとする課題】しかし、この方法で
は、時間の経過と共に細胞の生存率が低下してくるた
め、適当な時期に保存している細胞を融解し大量の増殖
させ、また凍結保存するという操作を繰り返さねばなら
ない。保存細胞種が多ければ細胞の維持管理は、膨大な
労力を要する作業となるため、長期にわたって細胞を凍
結しておける保存液が必要になって来る。However, in this method, the survival rate of cells decreases with the passage of time. Therefore, cells stored at an appropriate time are thawed to proliferate in a large amount, and also cryopreserved. You have to repeat the operation of doing. Since the maintenance of cells is a laborious task if there are many preserved cell types, a preservation solution that can freeze the cells for a long period of time is required.
【０００４】そこで、本発明者等は、上述の問題をでき
るかぎり解決する方法を鋭意検討した結果、細胞培養用
培地に、蛋白質、糖あるいは糖蛋白質からなる天然高分
子化合物やポリエチレングリコールなどの有機合成高分
子化合物、ジメチルスルホキシド、糖及び緩衝液を含む
保存液が、プログラムフリーザーを用いず、短時間に簡
単な操作で各種細胞を保存できる保存液を開発すること
ができた。Therefore, the inventors of the present invention have made extensive studies as to how to solve the above-mentioned problems as much as possible, and as a result, as a result, in a cell culture medium, organic substances such as proteins, sugars, natural polymer compounds composed of glycoproteins, polyethylene glycol, etc. A preservative solution containing a synthetic polymer compound, dimethylsulfoxide, sugar, and a buffer solution could be developed to save various cells by a simple operation in a short time without using a program freezer.
【０００５】[0005]
【課題を解決するための手段】本発明は、細胞培養用培
地に、天然高分子化合物または有機合成高分子化合物、
ジメチルスルホキシド、単糖または２糖及び緩衝液を含
むことを特徴とする細胞凍結保存液、及び天然高分子化
合物として、哺乳類由来血清５０〜９０％を含むことを
特徴とする細胞凍結保存液、及び天然高分子化合物とし
て、デキストラン０．１〜１．０％を含むことを特徴と
する細胞凍結保存液、及び天然高分子化合物として、グ
リコーゲン０．１〜１．０％を含むことを特徴とする細
胞凍結保存液、及び天然高分子化合物として、メチルセ
ルロースまたはカルボキシメチルセルロース０．１〜
１．０％を含むことを特徴とする細胞凍結保存液、及び
有機合成高分子化合物として、ポリエチレングリコール
１〜１０％を含むことを特徴とする細胞凍結保存液、及
び有機合成高分子化合物として、ポリビニルピロリドン
１〜１０％を含むことを特徴とする細胞凍結保存液、及
び単糖として、グルコース１〜１０％を含むことを特徴
とする細胞凍結保存液、及び２糖として、シュークロー
ス１〜１０％を含むことを特徴とする細胞凍結保存液、
に係るものである。The present invention provides a cell culture medium containing a natural polymer compound or an organic synthetic polymer compound,
A cell cryopreservation liquid containing dimethyl sulfoxide, a monosaccharide or a disaccharide, and a buffer, and a cell cryopreservation liquid containing 50 to 90% of mammalian-derived serum as a natural polymer compound, and A cell cryopreservation liquid containing 0.1 to 1.0% of dextran as a natural polymer compound, and 0.1 to 1.0% of glycogen as a natural polymer compound. As a cell cryopreservation solution and a natural polymer compound, methylcellulose or carboxymethylcellulose 0.1 to 0.1
As a cell cryopreservation liquid containing 1.0% and an organic synthetic polymer compound, as a cell cryopreservation liquid containing 1 to 10% polyethylene glycol, and an organic synthetic polymer compound, Cell cryopreservation liquid containing 1 to 10% of polyvinylpyrrolidone, and cell cryopreservation liquid containing 1 to 10% of glucose as a monosaccharide, and sucrose 1 to 10 as a disaccharide % Cell cryopreservation solution,
It is related to.
【０００６】[0006]
【作用】本保存液は、高分子化合物、ジメチルスルホキ
シド、細胞培養用培地、糖及び緩衝液の５成分からな
る。以下成分ごとに説明する。The preservative solution consists of five components: polymer compound, dimethyl sulfoxide, cell culture medium, sugar and buffer solution. Each component will be described below.
【０００７】１ 細胞培養用培地 基本的には、細胞培養が可能な基本培地であれば、どの
培地でも良く、オートクレーブができる培地の方が操作
及び滅菌という観点からより好適である。1 Medium for Cell Culture Basically, any medium can be used as long as it is a basic medium capable of culturing cells, and a medium capable of autoclaving is more preferable from the viewpoint of operation and sterilization.
【０００８】２ 高分子化合物 本保存液に適用される高分子化合物には、蛋白質、糖、
糖蛋白質等からなる天然高分子化合物と、エチレングリ
コールなどの有機合成化合物が考えられる。前者には各
種動物由来の血清、ゼラチン、アルブミン、肉エキス、
ミルク蛋白質、植物蛋白質、デンプン、デキストラン、
グリコーゲン、メチルまたはカルボキシメチルセルロー
ス、イヌリン、イノシトール及びソルビトール等が考え
られ、使用濃度は蛋白質や糖蛋白質では５０〜９０％、
糖の場合は０．１〜１０％で使用すると良好な結果が得
られる。また、後者には、ジエチレングリコール、プロ
ピレングリコール、ポリビニルピロリドン及びポリエチ
レングリコール等が上げられ、１〜１０％の濃度で使用
すると良い結果が得られる。2 Polymer Compound The polymer compound applied to the preservation solution includes proteins, sugars,
Natural polymer compounds such as glycoproteins and organic synthetic compounds such as ethylene glycol are considered. The former includes serum derived from various animals, gelatin, albumin, meat extract,
Milk protein, vegetable protein, starch, dextran,
Glycogen, methyl or carboxymethyl cellulose, inulin, inositol, sorbitol, etc. are considered, and the concentration used is 50 to 90% for proteins and glycoproteins,
In the case of sugar, good results are obtained when used at 0.1-10%. The latter includes diethylene glycol, propylene glycol, polyvinylpyrrolidone, polyethylene glycol and the like, and good results are obtained when used at a concentration of 1 to 10%.
【０００９】３ 糖 本保存液に使用される糖としては、グルコース、キシロ
ース、アラビノース、ガラクトース、フルクトース、ラ
クトース、シュークロース、マルトース、ラフィノース
等の単糖、２糖が細胞凍結保存液として良好な結果が得
られる。使用濃度は１０〜５０％が最も良い結果を示
す。Trisaccharides Monosaccharides such as glucose, xylose, arabinose, galactose, fructose, lactose, sucrose, maltose, and raffinose, and disaccharides, have good results as a cell cryopreservation solution. Is obtained. The best result is that the concentration used is 10 to 50%.
【００１０】４ ジメチルスルホキシド ジメチルスルホキシドは、一般的に使用されている凍結
保護剤で、使用濃度は５〜１５％が適当である。この他
に、グリセロールが使用できるが、本細胞凍結保存剤と
してはＤＭＳＯより効果は劣る。4 Dimethylsulfoxide Dimethylsulfoxide is a commonly used cryoprotective agent, and it is suitable to use it at a concentration of 5 to 15%. In addition to this, glycerol can be used, but it is less effective than DMSO as the present cell cryopreservative.
【００１１】５ 緩衝液 本保存液の水素イオン濃度は、室温において、７．０〜
８．５の範囲が適当である。これに使用されるものとし
ては基本的にはｐＨが上記の範囲内に収るものであれば
いずれの緩衝液でも良い。本保存液には、リン酸系、Ｂ
ＥＳ、ＴＥＳ、アセトアミドグリシン、ＴＲＩＣＩＮ
Ｅ、グリシンアミド、グリシルグリシン、ベロナール、
トリスエタノールアミン及びＨＥＰＥＳが使用可能で、
濃度は１００ｍＭ〜２００ｍＭが適当である。5 Buffer Solution The hydrogen ion concentration of this preservation solution is 7.0 to 7.0 at room temperature.
A range of 8.5 is suitable. Basically, any buffer solution may be used as long as it has a pH within the above range. This preservation solution contains phosphoric acid, B
ES, TES, acetamidoglycine, TRICIN
E, glycinamide, glycylglycine, veronal,
Trisethanolamine and HEPES can be used,
A suitable concentration is 100 mM to 200 mM.
【００１２】[0012]
【実施例及び試験例】以下、本発明の実施例及び試験例
を示して説明する。[Examples and Test Examples] Examples and test examples of the present invention will be described below.
【００１３】実施例１ 細胞凍結保存液の各成分を以下のように調製した。Example 1 Each component of the cell cryopreservation solution was prepared as follows.
【００１４】１）成牛血清（ヤガイ社製）７５０ｍｌ 原料試験（無菌試験、迷入ウイルス否定試験、エンドト
キシン測定）に合格した成牛血清を、５６℃、３０分温
水処理し、０．４５μＭプレフィルター及び０．２２μ
Ｍ無菌フィルター（ミリポア社製）を用いて滅菌した血
清を用いた。1) Adult bovine serum (manufactured by Yaigai Co., Ltd.) 750 ml Adult bovine serum that passed the raw material test (sterility test, stray virus negative test, endotoxin measurement) was treated with warm water at 56 ° C. for 30 minutes, and 0.45 μM prefilter And 0.22μ
Serum sterilized using an M sterilizing filter (Millipore) was used.
【００１５】２）ジメチルスルホキド１００ｍｌ ジメチルスルホキシド（和光純薬社製）１００ｍｌを
０．２２μＭ無菌フィルター（ミリポア社テフロンフィ
ルター）を用いて滅菌した。2) 100 ml of dimethyl sulfoxide 100 ml of dimethyl sulfoxide (manufactured by Wako Pure Chemical Industries, Ltd.) was sterilized using a 0.22 μM sterile filter (Teflon filter manufactured by Millipore).
【００１６】３）Ｂａｓｉｃ Ｓｔｏｃｋ Ｓｏｌｕｔ
ｉｏｎ（ＢＳＳ）１５０ｍｌ ＲＰＭＩ１６４０（Ｆｌｏｗ社製）１．５７５ｇ、グル
コース３０．０ｇ、炭酸水素ナトリウム０．８ｇ、ＨＥ
ＰＥＳ（和光純薬社製）０．３６ｇを二段蒸留水に加え
全量を１５０ｍｌとし、０．４５μＭプレフィルター
（ミリポア社製）０．２２μＭ無菌フィルター（ミリポ
ア社製）を用いて滅菌した。3) Basic Stock Solut
ion (BSS) 150 ml RPMI1640 (manufactured by Flow) 1.575 g, glucose 30.0 g, sodium hydrogen carbonate 0.8 g, HE
0.36 g of PES (manufactured by Wako Pure Chemical Industries, Ltd.) was added to double distilled water so that the total amount was 150 ml, and sterilized using a 0.45 μM prefilter (manufactured by Millipore) and a 0.22 μM sterile filter (manufactured by Millipore).
【００１７】４）調合 １）、２）、３）によって得た成牛血清７５０ｍｌ、ジ
メチルスルホキシド１００ｍｌ、Ｂａｓｉｃ Ｓｔｏｃ
ｋ Ｓｏｌｕｔｉｏｎ１５０ｍｌをクリーンベンチ内で
無菌的に調製した（ＣＢ−１）。4) Formulation 1) 750 ml of adult bovine serum obtained by 2), 3), 100 ml of dimethyl sulfoxide, Basic Stoc
150 ml of k Solution was aseptically prepared in a clean bench (CB-1).
【００１８】実施例２ 実施例１で調製した保存成分中で、ＲＰＭＩ１６４０に
代えて、１９９培地を用いて、実施例１と同様に保存液
を調製した（ＣＢ−２）。Example 2 A preservative solution was prepared in the same manner as in Example 1 by using 199 medium instead of RPMI1640 in the preservative components prepared in Example 1 (CB-2).
【００１９】実施例３ 実施例１の調製成分中、牛血清の代わりに、０．５％カ
ルボキシメチルセルロースを用いて、実施例１と同様に
保存液を調製した（ＣＢ−３）。Example 3 A preservative solution was prepared in the same manner as in Example 1 except that 0.5% carboxymethylcellulose was used instead of bovine serum in the preparation components of Example 1 (CB-3).
【００２０】実施例４ 実施例１の調製成分中、グルコースの代わりに、シュー
クロースを用いて、実施例１と同様の操作で保存液を調
製した（ＣＢ−４）。Example 4 A stock solution was prepared in the same manner as in Example 1 except that sucrose was used instead of glucose in the preparation components of Example 1 (CB-4).
【００２１】試験例１ あらかじめ、ＧＩＴ培地（日本製薬）で培養したＪｕｒ
ｋａｔ細胞（Ｈｕｍａｎ Ｔ Ｃｅｌｌ Ｌｅｕｋａｅ
ｍｉａ）及びＳＫ−００７細胞（ＨｕｍａｎＭｙｅｌｏ
ｍａ）を遠心で集め細胞数５×１０5〜５×１０6個／ｍ
ｌになるように、実施例１〜４で調製した各種保存液１
ｍｌを加えて懸濁し、２ｍｌ容のクライオチューブ（ヌ
ンク社製）に小分けした。２０〜３０分間予備凍結後、
−８０℃または−１９６℃で一定期間保存した。６ヶ月
間凍結保存したクライオチューブ入りの細胞液を３０〜
３７℃の微温湯中で軽く振りながら迅速に融解し、直ち
にそれぞれの細胞の至適増殖用培地（１０ｍｌ）で１回
洗浄後、生細胞をＥＫＤＳヘマサイトメーター（萱垣製
作所製）で計測した。結果は下記表１に示した。Test Example 1 Jur previously cultured in GIT medium (Nippon Pharmaceutical Co., Ltd.)
kat cells (Human T Cell Leukae
Mia) and SK-007 cells (HumanMyelo)
ma) is collected by centrifugation and the number of cells is 5 × 10 5 to 5 × 10 6 cells / m
1 of various preservatives prepared in Examples 1 to 4
ml was added and suspended, and the mixture was divided into 2 ml cryotubes (manufactured by Nunc). After pre-freezing for 20-30 minutes,
It was stored at -80 ° C or -196 ° C for a certain period. Cryotube-containing cell fluid stored frozen for 6 months
The cells were thawed quickly in 37 ° C. warm water with gentle shaking, immediately washed once with the optimal growth medium (10 ml) for each cell, and the viable cells were measured with an EKDS hemacytometer (Kayagaki Seisakusho). The results are shown in Table 1 below.
【００２２】[0022]
【表１】 [Table 1]
【００２３】試験例２ ＣＢ−１を用い、あらかじめ、供試細胞をその増殖に適
した培地を用いて培養し、遠心で集めた各種細胞を試験
例１と同様の操作によって１〜３０ヶ月間保存し、その
生存率を計測した。結果は下記表２に示した。Test Example 2 Using CB-1, test cells were cultured in advance in a medium suitable for their growth, and various cells collected by centrifugation were subjected to the same operation as in Test Example 1 for 1 to 30 months. It preserve | saved and the survival rate was measured. The results are shown in Table 2 below.
【００２４】[0024]
【表２】 [Table 2]
【００２５】試験例３ 本保存液、ＣＢ−１及び従来法を用い、あらかじめ供試
細胞をその増殖に適した培地を用いて培養し、遠心で集
めた各種細胞を試験例１と同様の操作によって６ヶ月間
保存し、その生存率を計測した。結果は下記表３に示し
た。Test Example 3 Using the preservative solution, CB-1 and the conventional method, test cells were previously cultured in a medium suitable for their growth, and various cells collected by centrifugation were subjected to the same operation as in Test Example 1. Was stored for 6 months, and the survival rate was measured. The results are shown in Table 3 below.
【００２６】[0026]
【表３】 [Table 3]
【００２７】[0027]
【発明の効果】本発明の細胞凍結保存液によれば、プロ
グラムフリーザーを用いることなく、短時間に簡単な操
作で細胞の保存を行なうことができ、しかも長期間凍結
保存しても高い生存率を保持する効果がある。According to the cell cryopreservation solution of the present invention, cells can be preserved by a simple operation in a short time without using a program freezer, and the survival rate is high even when cryopreserved for a long period of time. Has the effect of holding.
Claims (9)
たは有機合成高分子化合物、ジメチルスルホキシド、単
糖または２糖及び緩衝液を含むことを特徴とする細胞凍
結保存液。1. A cryopreservation solution for cells, characterized in that the cell culture medium contains a natural polymer compound or an organic synthetic polymer compound, dimethyl sulfoxide, a monosaccharide or a disaccharide, and a buffer solution.
清５０〜９０％を含む請求項１記載の細胞凍結保存液。2. The cell cryopreservation liquid according to claim 1, which contains 50 to 90% of mammalian-derived serum as a natural polymer compound.
０．１〜１．０％を含む請求項１記載の細胞凍結保存
液。3. The cell cryopreservation solution according to claim 1, which contains 0.1 to 1.0% of dextran as a natural polymer compound.
０．１〜１．０％を含む請求項１記載の細胞凍結保存
液。4. The cell cryopreservation liquid according to claim 1, which contains 0.1 to 1.0% of glycogen as a natural polymer compound.
ースまたはカルボキシメチルセルロース０．１〜１．０
％を含む請求項１記載の細胞凍結保存液。5. Methyl cellulose or carboxymethyl cellulose 0.1 to 1.0 as a natural polymer compound.
The cell cryopreservation liquid according to claim 1, which contains 10%.
レングリコール１〜１０％を含む請求項１記載の細胞凍
結保存液。6. The cell cryopreservation liquid according to claim 1, which contains 1 to 10% of polyethylene glycol as the organic synthetic polymer compound.
ルピロリドン１〜１０％を含む請求項１記載の細胞凍結
保存液。7. The cryopreservation solution for cells according to claim 1, which contains 1 to 10% of polyvinylpyrrolidone as the organic synthetic polymer compound.
む請求項１記載の細胞凍結保存液。8. The cell cryopreservation liquid according to claim 1, which contains 1 to 10% glucose as a monosaccharide.
を含む請求項１記載の細胞凍結保存液。9. Sucrose 1-10% as disaccharide
The cell cryopreservation liquid according to claim 1, which comprises:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4228001A JPH0646840A (en) | 1992-08-04 | 1992-08-04 | Solution for freezing and storing cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4228001A JPH0646840A (en) | 1992-08-04 | 1992-08-04 | Solution for freezing and storing cell |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0646840A true JPH0646840A (en) | 1994-02-22 |
Family
ID=16869626
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP4228001A Pending JPH0646840A (en) | 1992-08-04 | 1992-08-04 | Solution for freezing and storing cell |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0646840A (en) |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002046368A1 (en) * | 2000-12-04 | 2002-06-13 | Humantec Ltd. | Cell-preservation liquid and method of preserving cells by using the liquid |
| WO2006033429A1 (en) | 2004-09-24 | 2006-03-30 | Seiren Kabushiki Kaisha | Composition for cell frozen-storage |
| WO2007058308A1 (en) * | 2005-11-17 | 2007-05-24 | Nippon Zenyaku Kogyo Co., Ltd. | Aqueous solution for cell preservation |
| JP2011507530A (en) * | 2007-12-21 | 2011-03-10 | ビン エム．アブドゥラー，ジャファー アリ | Protein-free gamete and embryo handling media products and culture media products |
| JP2012166084A (en) * | 2005-06-21 | 2012-09-06 | Pervasis Therapeutics Inc | Method and composition for enhancing vascular access |
| WO2017061392A1 (en) * | 2015-10-05 | 2017-04-13 | メビオール株式会社 | Method for preservation of cell |
| CN106719604A (en) * | 2017-02-04 | 2017-05-31 | 上海尚维生物科技有限公司 | Pet peripheral blood directly freezes reagent and method |
| WO2018084228A1 (en) * | 2016-11-04 | 2018-05-11 | 国立大学法人東京大学 | Solution for cryopreservation of animal cells or animal tissues, cryopreserved product, and cryopreservation method |
| WO2018110159A1 (en) | 2016-12-14 | 2018-06-21 | 株式会社大塚製薬工場 | Mammalian cell cryopreservation liquid |
| WO2019026910A1 (en) * | 2017-07-31 | 2019-02-07 | 株式会社ツーセル | Composition for cryopreservation, method for producing cryopreserved material, cell preparation, method for producing cell preparation, and kit for cryopreservation |
| JP2020072668A (en) * | 2014-07-09 | 2020-05-14 | ジェネンテック， インコーポレイテッド | Ph adjustment for improving thaw restoration of cell bank |
| EP4050097A1 (en) | 2021-02-26 | 2022-08-31 | Nikkiso Co., Ltd. | Method of producing frozen renal cells and frozen renal cell |
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Cited By (22)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002046368A1 (en) * | 2000-12-04 | 2002-06-13 | Humantec Ltd. | Cell-preservation liquid and method of preserving cells by using the liquid |
| WO2006033429A1 (en) | 2004-09-24 | 2006-03-30 | Seiren Kabushiki Kaisha | Composition for cell frozen-storage |
| JP2012166084A (en) * | 2005-06-21 | 2012-09-06 | Pervasis Therapeutics Inc | Method and composition for enhancing vascular access |
| WO2007058308A1 (en) * | 2005-11-17 | 2007-05-24 | Nippon Zenyaku Kogyo Co., Ltd. | Aqueous solution for cell preservation |
| JPWO2007058308A1 (en) * | 2005-11-17 | 2009-05-07 | 日本全薬工業株式会社 | Aqueous solution for cell preservation |
| US8460926B2 (en) | 2005-11-17 | 2013-06-11 | Nippon Zenyaku Kogyo Co., Ltd | Aqueous solution for cell preservation |
| JP2014113166A (en) * | 2005-11-17 | 2014-06-26 | Nippon Zenyaku Kogyo Kk | Aqueous solution for cell preservation |
| JP5763288B2 (en) * | 2005-11-17 | 2015-08-12 | 日本全薬工業株式会社 | Aqueous solution for cell preservation |
| JP2011507530A (en) * | 2007-12-21 | 2011-03-10 | ビン エム．アブドゥラー，ジャファー アリ | Protein-free gamete and embryo handling media products and culture media products |
| JP2020072668A (en) * | 2014-07-09 | 2020-05-14 | ジェネンテック， インコーポレイテッド | Ph adjustment for improving thaw restoration of cell bank |
| JP2017070221A (en) * | 2015-10-05 | 2017-04-13 | メビオール株式会社 | Method for preserving cells |
| WO2017061392A1 (en) * | 2015-10-05 | 2017-04-13 | メビオール株式会社 | Method for preservation of cell |
| WO2018084228A1 (en) * | 2016-11-04 | 2018-05-11 | 国立大学法人東京大学 | Solution for cryopreservation of animal cells or animal tissues, cryopreserved product, and cryopreservation method |
| JPWO2018084228A1 (en) * | 2016-11-04 | 2019-09-26 | 国立大学法人 東京大学 | Solution for cryopreservation of animal cell or animal tissue, frozen product, and cryopreservation method |
| US11540507B2 (en) | 2016-11-04 | 2023-01-03 | The University Of Tokyo | Solution for cryopreservation of animal cells or animal tissues, cryopreserved product, and cryopreservation method |
| WO2018110159A1 (en) | 2016-12-14 | 2018-06-21 | 株式会社大塚製薬工場 | Mammalian cell cryopreservation liquid |
| KR20190087611A (en) | 2016-12-14 | 2019-07-24 | 가부시기가이샤오오쓰까세이야꾸고오죠오 | Mammalian cell freezing solution |
| US11889829B2 (en) | 2016-12-14 | 2024-02-06 | Otsuka Pharmaceutical Factory, Inc. | Mammalian cell cryopreservation liquid |
| CN106719604A (en) * | 2017-02-04 | 2017-05-31 | 上海尚维生物科技有限公司 | Pet peripheral blood directly freezes reagent and method |
| WO2019026910A1 (en) * | 2017-07-31 | 2019-02-07 | 株式会社ツーセル | Composition for cryopreservation, method for producing cryopreserved material, cell preparation, method for producing cell preparation, and kit for cryopreservation |
| JPWO2019026910A1 (en) * | 2017-07-31 | 2020-07-27 | 株式会社ツーセル | Cryopreservation composition, cryopreservation production method, cell preparation, cell preparation production method, cryopreservation kit |
| EP4050097A1 (en) | 2021-02-26 | 2022-08-31 | Nikkiso Co., Ltd. | Method of producing frozen renal cells and frozen renal cell |
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