JP3177713B2 - How to save the blood for transfusion or blood products - Google Patents

How to save the blood for transfusion or blood products

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JP3177713B2
JP3177713B2 JP11159992A JP11159992A JP3177713B2 JP 3177713 B2 JP3177713 B2 JP 3177713B2 JP 11159992 A JP11159992 A JP 11159992A JP 11159992 A JP11159992 A JP 11159992A JP 3177713 B2 JP3177713 B2 JP 3177713B2
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blood
transfusion
oxygen
storage
stored
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JPH05305123A (en
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暢 佐藤
直人 岡崎
和徳 川村
克己 日吉
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科学技術振興事業団
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【発明の詳細な説明】 DETAILED DESCRIPTION OF THE INVENTION

【0001】 [0001]

【産業上の利用分野】本発明は、輸血用血液または赤血球製剤(以下、単に血液という)の保存方法に関する。 The present invention relates to blood for transfusion or red cell product (hereinafter, simply referred to as blood) on preserving.

【0002】 [0002]

【従来の技術】従来、輸血用血液は、血液抗凝固剤とブドウ糖等を含む保存液(たとえばCPD液)を収容し滅菌した密封容器に採血され、そのまま冷蔵庫内において大気中で4〜6℃の温度で保存されている。 Conventionally, blood for transfusion is bled sealed container containing sterilized preservation solution containing anticoagulant and glucose or the like (for example, CPD liquid), as 4 to 6 ° C. in air in a refrigerator It is stored in the temperature. この状態で保存された血液の使用期限は日本では採血後3週間と定められている。 Expiration date of stored blood in this state are defined as 3 weeks after blood collection in Japan. 現在ではその使用期限を延長させる工夫が種々成されている。 The currently devised to extend the expiration date has been variously made. その一つにアメリカ合衆国などで認可され利用されている従来の保存液に加えてアデニン等の薬剤を添加する保存方法がある。 There is stored a method of adding an agent such as adenine in addition to the conventional preservation solution One to authorized in such United States have been used. また、本邦でも最近MAP液による赤血球濃厚液保存が4〜6℃で6週間まで認可された。 In addition, red blood cell concentrates stored by the recent MAP solution also in Japan has been authorized up to 6 weeks at 4~6 ℃. しかし、これらの方法では長期にわたる保存のために生じる溶血、すなわち、赤血球中のヘモグロビンが赤血球の破壊ないしは赤血球膜の変化により赤血球より漏出してくることによって生じる血漿中遊離ヘモグロビン濃度の上昇を防ぐことはできない。 However, hemolysis occurs due to long-term preservation by these methods, i.e., preventing an increase in plasma free hemoglobin concentration caused by hemoglobin in red blood cells coming leaked from red blood cells by a change in fracture or erythrocyte membrane of red blood cells can not. また、 Also,
この方法は、アデニンなどの添加物の余剰分またはその代謝産物であるヒポキサンチンが、血液の生体注入の後に副作用を引き起こすこともあり、改良すべき点を残している。 This method, surplus or hypoxanthine its metabolites of additives such as adenine, sometimes cause side effects after the biological implantation blood, leaving points to be improved.

【0003】また、血液の凍結温度は約−2℃であり、 [0003] freezing temperature of the blood is about -2 ° C.,
血液の保存温度をこの血液の凍結前まで下げて保存する方法がある。 There is a method of saving by lowering the storage temperature of the blood before freezing of the blood. この方法は、生体注入時の副作用は少なく保存期間の延長を可能とするが、低温による障害に伴う血漿中の遊離ヘモグロビンの増加(溶血)が観察される。 This method, side effects during biological implantation allows for extension of less shelf life, but the increase in free hemoglobin in plasma due to the failure by low-temperature (hemolysis) is observed. この点に関して、本発明者らは、先に、現在臨床的に輸液点滴薬の形で多用されているグリセリンとマニト−ルを夫々少量づつ添加して、この低温による溶血障害ならびに赤血球内外のナトリウム、カリウム等の電解質の分配異常を防止、是正する方法を開発した(特公平2 In this regard, the present inventors have previously, glycerin and Manitowoc that is frequently used in the form of current clinical infusion drops - Le added respectively in small portions, sodium hemolysis disorders and erythrocytes out by the low-temperature , prevent the dispensing abnormality of electrolytes such as potassium, we developed a method to correct (KOKOKU 2
−62534)が、なかには稀にその効果が予想されたほどに期待できない低温障害に弱い血液も認められるために、その原因の追及ならびに改良を重ねてきた。 -62534) is to weak blood also found in cold disorders Nakaniwa rarely the effect can not be expected to more expected, it has been over pursuit and improvement of its cause.

【0004】 [0004]

【発明が解決しようとする課題】本発明者等は、上述の低温障害の主な原因を低温にするほど多くなる血漿中、 The present inventors have [0005] during so many made plasma to a main cause of low-temperature failure described above in a low temperature,
血液中の溶解酸素、特に活性酸素による障害との知見を得た。 Dissolved oxygen in the blood, in particular to obtain a knowledge that damage by active oxygen. すなわち、生体内を循環している血液は、肺で酸素濃度を高められ活性酸素の曝露を受けるが、血液は自身で保持している活性酸素に対する生理的防御機能により、また、組織では酸素濃度自体が低下することによって酸素による障害が通常(健康状態)では余り起こらないようになっている。 That is, blood that circulates in vivo is subjected to exposure of active oxygen is increased oxygen concentration in the lungs, the physiological defense against active oxygen blood held by itself, also, the oxygen concentration in the tissue failure by oxygen is prevented occur less in normal (healthy state) by itself decreases. しかし、低温状態で気体の溶解度が増加する現象は保存血液に関しても例外ではなく、血液中の酸素濃度は低温に成るほど増加する。 However, the phenomenon that the solubility of the gas at low temperature is increased no exception with respect stored blood, the oxygen concentration in the blood increases as temperature. しかし、酸素に曝露される赤血球などの組織には、常に、SOD However, organizations such as red blood cells from exposure to oxygen, always, SOD
(superoxide dismutase)やカタラ−ゼなどの活性酸素除去酵素が多量に含まれており、 (Superoxide Dismutase) and catalase - active oxygen removing enzyme such as peptidase is included in a large amount,
保存血液もこれらの働きを得るものとして酸素による障害はほとんど無視されたいた。 Stored blood is also oxygen-failure as to obtain these works were should be largely ignored.

【0005】ところが、本発明者等は、保存温度を従来よりも下げた凍結前領域(0±2℃)での保存を研究、 [0005] However, the present inventors have studied the storage at freezing front region having a reduced storage temperature than the conventional (0 ± 2 ℃),
具体的には0±1℃の領域で実験を実施した過程で、生体内血液と異なり酸素を含んでいる必要のない保存血液が保存期間中常に酸素を異常に多く含んだ状態に置かれていることを見出し、SODなどの酵素の働きも低温になるほど低下することから従来の保存方法では酸素による害を防止しきれないのではないかと考えた。 In specific process Experiments were performed in the region of 0 ± 1 ° C. The, it is placed do not need stored blood is always oxygen during storage that contains oxygen unlike vivo blood abnormally laden state It found that there was thought that it would be not be prevented harm by oxygen in the conventional preservation method since it decreases as it becomes cold working of enzymes, such as SOD. そこで、 there,
本発明者は、この酸素による障害を防ぐ手段として原因物質である保存中の酸素の濃度を他のガスを用いて積極的に低下させたところ、長期の保存に耐えることを見出し、本発明を完成したもので、本発明の目的は輸血用血液または赤血球製剤の長期にわたる液状保存方法を提供するにある。 The present inventors have revealed that actively reduce the concentration of oxygen in the causative agent is being saved as a means of preventing damage by oxygen with other gases, found to withstand prolonged storage, the present invention those completed, object of the present invention to provide a long-term liquid storage method of blood for transfusion or red blood cell products.

【0006】 [0006]

【課題を解決するための手段】本発明の要旨は、輸血用血液または血液製剤を亜酸化窒素ガス、若しくは、 亜酸 Gist of the present invention SUMMARY OF THE INVENTION may, nitrous oxide gas blood for transfusion or blood products, or, Asan
化窒素ガスと窒素との混合ガスの雰囲気下で保存することを特徴とする輸血用血液または血液製剤の保存方法であり、その際、輸血用血液または血液製剤をガス透過性の良い容器内に収容し、亜酸化窒素ガス若しくは亜酸化 Of a method of storing blood for transfusion or blood products, characterized in that stored in an atmosphere of a gas mixture of nitrogen gas and nitrogen, in which the blood for transfusion or blood products in the gas permeable good vessel accommodated, nitrous oxide, nitrogen gas or nitrous oxide
窒素ガスと窒素との混合ガスの雰囲気下で保存することが好ましく、更に、採血後、早期に輸血用血液または血液製剤に含まれている酸素を下げるために雰囲気(環境) を亜酸化窒素ガス若しくは亜酸化窒素ガスと窒素と It is preferable to store in an atmosphere of a gas mixture of nitrogen gas and nitrogen, further blood after early blood for transfusion or atmosphere to reduce the oxygen contained in the blood product (environment) the nitrous oxide gas or nitrous oxide gas and nitrogen
のガスでパ−ジすることが好ましい。 It is di preferred - path in the gas.

【0007】すなわち、本発明においては、保存中血液の酸素障害を低減させるために、血液中の活性酸素を減らすように空気を亜酸化窒素ガス若しくは亜酸化窒素ガ [0007] That is, in the present invention, in order to reduce the oxygen fault storage in the blood, nitrous oxide gas air to reduce the active oxygen in the blood or nitrous oxide gas
スと窒素との混合ガスで置換し、血液の酸素濃度を低下させて血液を保存するものであり、また、採血時または採血直後に亜酸化窒素若しくは亜酸化窒素と窒素との混合ガスを血液保存容器中あるいは血液保存容器の雰囲気中に急速に大量に供給して、容器中あるいはその雰囲気中の空気をパ−ジし、ついで、持続的に亜酸化窒素若しくは亜酸化窒素と窒素との混合ガスを添加して大気中の酸素が保存血液容器内に拡散浸透してくるのを防ぐ方法を提供するものである。 It was replaced with a mixed gas of scan and nitrogen, to lower the oxygen concentration of the blood is intended to store the blood and a gas mixture of nitrous oxide or nitrous oxide and nitrogen immediately after blood collection or during blood collection Blood rapidly large amount supplied to the atmosphere in the storage vessel or the blood storage container, the air or in the atmosphere in the vessel path - mixing and di, then the persistently nitrous oxide or nitrous oxide and nitrogen was added to the gas is to provide a method to prevent the oxygen in the air come to cementation in stored blood vessel.

【0008】次に、本発明について詳細に説明する。 [0008] Next, the present invention will be described in detail. 本発明において輸血とは単に他人からの血液の供給に止まらず、手術前の自己血液の貯血からの供給をも言うのであり、本発明において使用する亜酸化窒素とは笑気ガスとも称されるものであって、該笑気ガスを用いる場合には、その麻酔作用による細胞膜の安定化、水分子との親和性により低温での氷晶形成による障害を予防するなどの作用も期待でき、特に血液の低温保存に関して効果がよい。 Without stopping to simply supply of blood from another person and the blood transfusion in the present invention, it is to say also the supply from blood of preoperative autologous blood, also called laughing gas and nitrous oxide for use in the present invention be those, in the case of using a 該笑 air gas can be expected effects such as the stabilization of cell membranes by anesthetic action, the affinity for water molecules to prevent damage due to ice crystal formation at low temperatures, in particular good effect with respect to low-temperature preservation of blood. また、本発明において血液を保存する容器としては、ガス透過性の良い容器が好ましく、該容器中に血液を収容し、これを亜酸化窒素若しくは亜酸化窒素と窒素 As the container for storing blood in the present invention preferably has a good vessel gas permeability, accommodates the blood in the vessel, which nitrous oxide or nitrous oxide and nitrogen
の混合ガスで充満している雰囲気容器内に設置することによって、血液を収容した容器外から亜酸化窒素を拡散浸透させ、保存中の血液酸素濃度の増加、及び、大気中より血液内に浸透してくる酸素を抑制することができる。 By placing the filling to that atmosphere within the container with a gas mixture of blood is diffused penetrated the vessel or outside et nitrous oxide accommodate an increase in blood oxygen concentration in the saved, and the blood from the atmosphere it is possible to suppress the oxygen coming to penetrate within.

【0009】本発明において血液を保存するに適したガス透過性の良い容器とは0.02〜0.4mm程度の膜厚を有するポリ塩化ビニ−ル、ポリオレフィン、ポリシリコン等のプラスチックフイルムまたはシ−トで構成されているのであって、例えば、特公昭63−44374 [0009] polyvinyl chloride is a gas permeable good container suitable for storing blood in the present invention have a thickness of about 0.02~0.4Mm - le, polyolefin, or a plastic film such as polysilicon sheet - a than is constituted by preparative, for example, JP-B-63-44374
号や特公平1−42214号に記載されている容器が好適である。 Is suitable containers described in JP and Kokoku No. 1-42214. そして、これらの容器のガス透過率は、約6 Then, the gas permeability of these containers, about 6
500ml/m 2・24hr・atm前後である。 It is 500ml / m 2 · 24hr · atm before and after. 保存血液の保存温度としては、−2℃〜+6℃程度の範囲、 The storage temperature of the stored blood, -2 ℃ ~ + 6 ℃ range of about,
好ましくは、0℃±1℃の範囲である。 Preferably, in the range of 0 ℃ ± 1 ℃. また、保存環境中の血液の炭酸ガス濃度を調整、コントロ−ルすることによって保存中の血液pH指数を、血液の代謝にとって好ましい状態に保つことができる。 Further, the carbon dioxide concentration of the blood storage environment adjustment, control - Blood pH index during storage by Le, can be kept in a preferable state for blood metabolism. 通常、保存中の血液の炭酸ガス濃度としては、保存1週間以後85mmHg Normally, the carbon dioxide concentration in the blood during storage, storage for one week after 85mmHg
以下であり、保存中の血液pH指数としては、6.8〜 Or less, as the blood pH index during storage, 6.8
7.5の範囲にあることが好ましい。 It is preferably in the range of 7.5.

【0010】次に、実施例をもって更に具体的に本発明を説明する。 [0010] will now be described concretely present invention with examples.

【実施例】対象血液は6名の健康成人男子から、CPD [Example] target blood from a healthy adult male of six, CPD
液にグリセロ−ルとマニト−ル(最終濃度はそれぞれ0.5、0.125W/V%)を含んだ膜厚380μm Glycero liquid - le and Manitowoc - Le thickness containing (final concentrations are 0.5,0.125W / V%) 380μm
医療用のPVC製通常容器と、膜厚130μmの医療用PVC製薄膜容器とに、1名につき200mlずつ計4 And PVC made ordinary container for medical use, in the medical PVC thin film made of container with a thickness of 130μm, each 200ml per person a total of 4
00ml採血した。 00ml blood was drawn. なお通常容器と薄膜容器の採血順序は交互に替えて行った(クロスオ−バ−デザイン)。 Note bled order of normal containers and films vessel went instead alternating (cross over - bar - Design). 採血当日の検査を行った後、薄膜容器血液は全容9lの金属製容器に入れて笑気ガスで速やかに金属製容器内の空気を置換した。 After inspection of the day of the blood collection, the thin film vessel blood to replace the air in the rapidly metallic vessel with laughing gas placed in a metal container having an overall 9l. 薄膜容器血液はこの金属製容器を4〜6 Film container Blood the metal container 4-6
ml/分の笑気ガスで換気しながら、通常容器血液(対照群)と共に1±1℃に保たれた同じ冷蔵庫内で5週間保存して比較を行った。 While ventilation ml / min laughing gas, in normal vessels blood (control group) the same in a refrigerator kept at 1 ± 1 ° C. with 5 weeks of storage was performed comparison. なお換気に用いられた笑気ガスは冷蔵庫外に直接排出することにより対照群に影響を及ぼさないようにしてある。 Incidentally laughing gas used in ventilation are in order not to affect the control group by discharging directly to the outside of the refrigerator. 測定結果の一部を表1に示す。 Some results are shown in Table 1. なお、測定については、Student's pa It should be noted that, for the measurement, Student's pa
ired t−testの統計的検定を用いた。 Using a statistical test of ired t-test. 危険率;p<0.05として行った。 It was carried out with p <0.05; hazard ratio.

【0011】 [0011]

【表1】 [Table 1]

【0012】表1において、括弧内は単位、数値は平均値および標準誤差(mean±SD)を表す。 [0012] In Table 1, in parentheses units, numbers represent the mean and standard error (mean ± SD). Stud Stud
ent's paired t−testを用いた統計的検定の結果、5週間目の値は何れの項目についても対照群との間に危険率5%以下で(p<0.05)有意な差が認められた。 ent's paired t-test result of the statistical test using, the (p <0.05) significantly different risk rate of 5% or less between the control group for 5 weeks values ​​any item Admitted. すなわち、本発明による保存方法は、対照群と比べ酸素濃度の増加を抑制することにより溶血と赤血球細胞膜の透過性の異常を抑え、また、グルコ−ス値の低下や乳酸値の上昇といった変化は代謝がより盛んに行われたことを示し、本来なら血液pHが代謝量に相当して低下するべきところであるにも拘らず、現行の3週間保存血液の平均pH6.6よりもはるかに高く保たれており、さらに、電解質バランスもより好ましい状態で保存されたことを示しているもので、本発明の優位性を示すものである。 That is, the method storage according to the invention, to suppress the abnormal permeability of hemolysis and red blood cell membranes by inhibiting the increase in oxygen concentration as compared with the control group, also, gluco - changes such increase in reduction and lactic acid value of the scan values metabolism show that took place more actively, despite is where should the blood pH would normally decreases correspond to metabolic rate, much higher coercive than the average pH6.6 current three weeks stored blood sauce and, further, electrolyte balance be those indicating that stored in a more preferred state, illustrates the superiority of the present invention.

【0013】 [0013]

【発明の効果】以上述べたように、本発明によれば保存中の溶血にともなう遊離ヘモグロビンの増加を抑制することができ、従来よりも低い温度での血液保存を可能にするもので、現行保存法よりも保存血液の質の向上を長期間にわたって維持することができる。 As described above, according to the present invention, according to the present invention it is possible to suppress an increase in free hemoglobin due to hemolysis during storage, one that enables the blood storage at lower than conventional temperatures, current than storage method can be maintained over a long period of time to improve the quality of stored blood. したがって本発明による血液保存は貴重な保存血液を無駄にすることを少なくする。 Thus blood storage according to the invention to reduce the wasting valuable stored blood. また、自己血の術前貯血期間を長くすることにより、良質の自己血を貯えやすくして他人からの血液を使うに伴う輸血の副作用、合併症の減少に役立てることができる。 Further, by increasing the preoperative blood period autologous blood can serve transfusion side effects associated with with easily stored and autologous blood quality using blood from another person, the reduction of complications.

フロントページの続き (56)参考文献 特開 平2−74257(JP,A) 特開 昭59−33224(JP,A) 特開 昭63−60931(JP,A) (58)調査した分野(Int.Cl. 7 ,DB名) A61J 3/00 A61K 35/14 Of the front page Continued (56) Reference Patent flat 2-74257 (JP, A) JP Akira 59-33224 (JP, A) JP Akira 63-60931 (JP, A) (58) investigated the field (Int .Cl. 7, DB name) A61J 3/00 A61K 35/14

Claims (3)

    (57)【特許請求の範囲】 (57) [the claims]
  1. 【請求項1】 輸血用血液または血液製剤を亜酸化窒素ガス、若しくは、 亜酸化窒素ガスと窒素との混合ガスの雰囲気下で保存することを特徴とする輸血用血液または血液製剤の保存方法。 [Claim 1] The blood for transfusion or blood products nitrous oxide gas, or storage method blood for transfusion or blood products, characterized in that stored in an atmosphere of a gas mixture of nitrous oxide gas and nitrogen.
  2. 【請求項2】輸血用血液または血液製剤をガス透過性の良い容器内に収容し、亜酸化窒素ガス、若しくは、 亜酸 Wherein the blood for transfusion or blood products containing the gas permeable good vessel, nitrous oxide gas, or, Asan
    化窒素ガスと窒素との混合ガスの雰囲気下で保存することを特徴とする輸血用血液または血液製剤の保存方法。 The storage method blood for transfusion or blood products, characterized in that stored in an atmosphere of a gas mixture of of nitrogen gas and nitrogen.
  3. 【請求項3】採血後、早期に輸血用血液または血液製剤の雰囲気ガスを亜酸化窒素ガス若しくは亜酸化窒素ガス 3. A blood collection after early nitrous oxide gas ambient gas blood for transfusion or blood products or nitrous oxide gas
    と窒素との混合ガスで空気又は酸素をパ−ジすることを Pas air or oxygen in a mixed gas of nitrogen - and di child
    特徴とする輸血用血液または血液製剤の保存方法。 The storage method blood for transfusion or blood products, wherein.
JP11159992A 1992-04-30 1992-04-30 How to save the blood for transfusion or blood products Expired - Fee Related JP3177713B2 (en)

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CN102711865B (en) 2009-10-12 2015-08-19 新健康科学股份有限公司 Blood storage bag system and the consuming device having an oxygen and carbon dioxide consumption capacity
CA2781874A1 (en) 2009-10-12 2011-04-21 New Health Sciences, Inc. Oxygen depletion devices and methods for removing oxygen from red blood cells
US9199016B2 (en) 2009-10-12 2015-12-01 New Health Sciences, Inc. System for extended storage of red blood cells and methods of use
US9339025B2 (en) 2010-08-25 2016-05-17 New Health Sciences, Inc. Method for enhancing red blood cell quality and survival during storage
JP5859558B2 (en) * 2010-11-05 2016-02-10 ニュー・ヘルス・サイエンシーズ・インコーポレイテッドNew Health Sciences, Inc. Save irradiation and anaerobic erythrocytes
US9067004B2 (en) 2011-03-28 2015-06-30 New Health Sciences, Inc. Method and system for removing oxygen and carbon dioxide during red cell blood processing using an inert carrier gas and manifold assembly
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CA2978940A1 (en) 2015-03-10 2016-09-15 New Health Sciences, Inc. Oxygen reduction disposable kits, devices and methods of use thereof
EP3285711A4 (en) 2015-04-23 2018-11-07 New Health Sciences, Inc. Anaerobic blood storage containers

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