JPS5995051A - Apparatus for fractionating and purifying blood - Google Patents
Apparatus for fractionating and purifying bloodInfo
- Publication number
- JPS5995051A JPS5995051A JP58204243A JP20424383A JPS5995051A JP S5995051 A JPS5995051 A JP S5995051A JP 58204243 A JP58204243 A JP 58204243A JP 20424383 A JP20424383 A JP 20424383A JP S5995051 A JPS5995051 A JP S5995051A
- Authority
- JP
- Japan
- Prior art keywords
- blood
- adsorbent
- column
- red blood
- blood cells
- Prior art date
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Abstract
(57)【要約】本公報は電子出願前の出願データであるた
め要約のデータは記録されません。(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.
Description
【発明の詳細な説明】
本発明は血液中から白血球・白小板を除去して赤血球を
分画・精製する装置に関するもので、特に成分輸血ある
いは生化学検査において、白血球・血小板を含まない赤
血球を得るための血液の分画・精製装置を提供すること
を目的とするものである。Detailed Description of the Invention The present invention relates to an apparatus for separating and purifying red blood cells by removing white blood cells and platelets from blood. In particular, the present invention relates to a device for separating and purifying red blood cells by removing white blood cells and platelets from blood. The purpose of this invention is to provide a blood fractionation/purification device for obtaining blood.
従来、輸血においては全血を用いる場合が多かったが、
最近は医療の進歩に併っていわゆる成分輸血と呼ばれる
必要な成分だけを輸血する治療法が国内外において進め
られている。然るに赤血球の成分輸血において、HL−
A型抗原を有する白血球および血小板抗原を有する血小
板を完全罠除去する優れた方法がないために、必ずしも
赤血球輸血が頻繁に行なわれているとは限らない。また
、赤血球の生化学的機能試験においても他の血球成分の
混在は良い結果をもたらさない。Traditionally, whole blood was often used in blood transfusions, but
Recently, with advances in medical care, a therapeutic method called component transfusion, in which only the necessary components are transfused, is being promoted both domestically and internationally. However, in red blood cell component transfusion, HL-
Red blood cell transfusions are not always performed frequently because there is no good way to completely remove leukocytes with type A antigens and platelets with platelet antigens. Also, in a biochemical function test of red blood cells, the presence of other blood cell components does not yield good results.
輸血を目的とした赤血球の分画法の公知技術としては次
のような方法がある。すなわち、(1)到立遠心分離法
、(2)生理食塩水洗浄法、(3)デキストランまたは
HESによる赤血球沈降法、(4)ナイロンカラム通過
法、(5)解凍赤血球浮遊液の調製〔ブラッド コンポ
ーネント テエラピー、アメリカンアソシエーション
オプ ブラッド バンクス刊。Known techniques for fractionating red blood cells for the purpose of blood transfusion include the following methods. Namely, (1) centrifugation method, (2) saline washing method, (3) erythrocyte sedimentation method using dextran or HES, (4) nylon column passage method, (5) preparation of thawed red blood cell suspension [blood cell suspension method]. Component Therapy, American Association
Published by Op Blood Banks.
ツウエンティース センチェリーフレスU、 8. A
(Blood Component Therapy
、 AmericanAs5ociation of
Blood Banks 、 Twentyth Ce
nturyPress U、 S、 A ) 、 9−
13頁1969年;日本赤十字崩液センター業務規準
昭和50年;血液成分輸血時代要覧 大阪赤十字センタ
ー刊 昭和50年〕があり、この中で(3)は白血球除
去赤血球「日歩」として実際に行なわれているが、いず
れも赤血球の回収および白血球・血小板の除去に関して
は満足されるものではない。また上記(4)の如き吸着
体をカラムにつめて行なう方法として木綿および木綿糸
を用いる方法〔フレミング(Fleming ) +ブ
リティッシュ ジャーナル オプ エキスペリメンタル
パソロジー(13ritish Journal of
ExperimentalPathalog)’)、
7巻281頁、 1926年;デイペンホルスト(pi
epenhorst )ら、ボックスサイクニス(Vo
x 3aug、) 、 23巻 30B −320頁、
1972年〕。Twentie's Century Fres U, 8. A
(Blood Component Therapy
, AmericanAs5organization of
Blood Banks, Twentyth Ce
nturyPress U, S, A), 9-
Page 13 1969; Japanese Red Cross Collapse Center Business Standards
1975; Handbook of the Era of Blood Component Transfusion, published by Osaka Red Cross Center, 1975], in which (3) is actually performed as leukocyte-removed red blood cell "Nippo", both of which involve collection of red blood cells and white blood cell collection. - Removal of platelets is not satisfactory. In addition, as a method of packing an adsorbent in a column as described in (4) above, there is a method using cotton and cotton thread [Fleming + British Journal of Experimental Pathology (13ritish Journal of Experimental Pathology)].
Experimental Pathalog)'),
Vol. 7, p. 281, 1926; Diepenhorst (pi
epenhorst) et al., Boxcycnis (Vo
x 3aug, ), vol. 23, p. 30B-320,
1972].
ダヌロン繊維を用いる方法〔ラング7エルダー(Lan
gfelder )ら、ボックス サイグイニス(Vo
x3ang、) + 19巻57頁、 1970年〕が
あるが、血小板や白血球の中のリンパ球の分離が良好で
ないし、血液量に対し吸着体の量が多く赤血球の回収が
充分行なわれない。Method using Danelon fiber [Lang 7 Elder (Lan
gfelder) et al., Box Saiguinis (Vo
x3ang, ) + Vol. 19, p. 57, 1970], but the separation of lymphocytes from platelets and white blood cells is not good, and the amount of adsorbent is large compared to the amount of blood, so red blood cells cannot be recovered sufficiently.
中量ら〔ネイチャー ニュー バイオロジー(Natu
re New Biology ) 、 246巻 9
4頁、 1973年〕はヘパリン加全通から陽イオン交
換セルロースや陽イオン交換セファデックスを用いて赤
血球を純化しているが、イオン交換体が高価であり又マ
グネシウムイオンが不可決であること、そして血液量に
対し吸着体の量が多い等の欠点があり実用的でない。一
方ライト(Wright ) [ザランセット(The
Lancet ) + 1巻4頁、 1926年〕は
ろ紙に白血球が吸着することをみとめているが、ガルビ
ン(Garvin ) (ジャーナル オプ エキスベ
リメンタル メデイシy (Journal of E
xperlmentalMediaine ) s 1
14巻51頁、 1961年〕はろ紙に対する吸着はさ
ほど大きくないとしている0本発明者は上記の事情を鑑
み鋭意研究した結果、ヒト全血、赤血球流層および哺乳
動物の血液からセルロース粉末、結晶セルロース、陰イ
オン交換セルロースに殆んど完全に白血球・血小板を付
着せしめて赤血球を高純度、かつ高収率で回収すること
に成功し本発明を完成するに至った〇そして本発明の目
的を達する構成は、カラムと該カラム内に吸着体を有す
る血液の分画・精製装置において、前記カラムは血液流
入口と血液流出口を有し、カラム内にはセルロース粉末
、結晶セルロース、陰イオン交換セルロースのうち少な
くともひとつからなる吸着体を充填し、さらに、該吸着
体を支持するメツシュが設けられていることを特徴とす
る血液の分画・精製装着。Nakamura et al. [Nature New Biology (Natsu)
re New Biology), Volume 246, 9
4, 1973] used cation-exchanged cellulose or cation-exchanged Sephadex to purify red blood cells from a heparinized solution, but ion exchangers were expensive and magnesium ions were not available. Moreover, it has drawbacks such as the amount of adsorbent being large relative to the amount of blood, making it impractical. On the other hand, Wright [The Lancet]
Lancet) + Vol. 1, p. 4, 1926] found that white blood cells adsorbed to filter paper, but Garvin (Journal of Experimental Medicine)
xperlmentalMediaine) s 1
Vol. 14, p. 51, 1961] states that adsorption to filter paper is not very large. In view of the above circumstances, the present inventor conducted intensive research and found that cellulose powder and crystals can be extracted from human whole blood, red blood cell flow layer, and mammalian blood. They succeeded in recovering red blood cells with high purity and high yield by almost completely adhering white blood cells and platelets to cellulose and anion-exchanged cellulose, and completed the present invention. The configuration achieved is a blood fractionation/purification device having a column and an adsorbent in the column, the column has a blood inlet and a blood outlet, and the column contains cellulose powder, crystalline cellulose, anion exchanger, etc. A device for fractionating and purifying blood, characterized in that it is filled with an adsorbent made of at least one of cellulose and is further provided with a mesh that supports the adsorbent.
本発明の実施において用いる血液としては、ヒト新鮮血
にヘパリン、 ACD 、 CPD等の適当な抗凝固剤
を加えたものおよび赤血球流層(日歩)であるが、必ず
しもこれに限定されるものではなく赤血球が浮遊した製
剤ならなんでもよい。赤血球流層を用いる場合は、生理
食塩水又は等張のバッファーを1/3〜1/2容加えて
用いる。The blood used in the practice of the present invention includes human fresh blood to which an appropriate anticoagulant such as heparin, ACD, CPD, etc. has been added, and red blood cell flow layer (Nippo), but is not necessarily limited thereto. Any preparation that has red blood cells suspended in it is fine. When using a red blood cell flow bed, add 1/3 to 1/2 volume of physiological saline or isotonic buffer.
また、本発明の実施において用いる吸着体としては、セ
ルロース粉末、結晶セルロース、陰イオン交換セルロー
スであシ、例えば山陽国策パルプ製KCフロック、東洋
ろ紙製ろ紙粉末、旭化成工業製アビセル、ジエチルアミ
ノエチルセルロースなどを挙げることができる。ジエチ
ルアミノエチルセルロースの如き陰イオン交換体はOH
−型で用いてもよく、予じめCI−型として用いてもよ
いが安定性を考慮して塩型の方が好ましい。In addition, adsorbents used in the practice of the present invention include cellulose powder, crystalline cellulose, anion exchange cellulose, such as KC floc manufactured by Sanyo Kokusaku Pulp, filter paper powder manufactured by Toyo Roshi, Avicel manufactured by Asahi Kasei Corporation, diethylaminoethyl cellulose, etc. can be mentioned. Anion exchangers such as diethylaminoethyl cellulose are OH
Although it may be used in the - form or may be used in advance as the CI- form, the salt form is preferable in consideration of stability.
これらの吸着体は生理食塩水で予じめ洗浄して用いる。These adsorbents are washed in advance with physiological saline before use.
本発明の実施において前記吸着体をポリプロピレン製等
の硬質ブ2スナックでできたカラムに25ミクロン程度
のオープニングを有するメツシュ、例えばナイロンメツ
シュで支持する吸着体のカラムへの充填は、生理食塩水
KWIA濁した吸着体を流しこみ、場合によシ加圧して
2〜6td/19の割でつめる。吸着体の量は、血液1
00−に対して10〜50−の範囲である。吸着体に血
液を通したあとの洗浄は、赤血球の回収率をあげるため
に行なった方がよく、洗浄液は生理食塩水で行なう。In carrying out the present invention, the adsorbent is supported by a mesh having an opening of about 25 microns, such as a nylon mesh, in a column made of a hard plastic snack made of polypropylene, etc., and the adsorbent is filled into the column with physiological saline. Pour in the KWIA cloudy adsorbent, pressurize if necessary, and pack at a rate of 2 to 6 td/19. The amount of adsorbent is blood 1
It ranges from 10-50- to 00-. Washing after passing blood through the adsorbent is recommended to increase the recovery rate of red blood cells, and the washing solution should be physiological saline.
洗浄の生理食塩水の量は吸着体1−に対し1〜3−が適
当である0カラムを通過せしめる時間は5〜120分の
範囲で、流速が遅い場合はIV4/−以下の力で加圧し
て溶出を早めてもよい〇以下の実施例1〜5に示される
ように、本発明の方法によってヒト全血、赤血球流層お
よび哺乳動物の血液から赤血球を分画した結果、赤血球
の回収率は95%以上白崩球、血小板の除去率は99チ
以上という結果が得られた。回収された赤血球を走査型
電子顕微鏡で観察すると吸着体に接触する前と全く同一
の円盤状を示し、パーパート法で浸透圧脆弱性とみると
吸着体接触前後で全く変化は ゛なかった。又、吸着体
に接触させたものとさせなかったものとを4℃で1週間
保存後、アデノシン−トリリン酸、2.3−ジホスホグ
リセン酸の定量を打力った結果、定量値に全く差はみと
められなかった。The appropriate amount of physiological saline for washing is 1 to 3 to 1 of the adsorbent.The time for passing through the column is 5 to 120 minutes, and if the flow rate is slow, apply with a force of IV4/- or less. As shown in Examples 1 to 5 below, as a result of fractionating red blood cells from human whole blood, red blood cell flow layer, and mammalian blood by the method of the present invention, the recovery of red blood cells Results showed that the removal rate of white blood cells was over 95%, and the removal rate of platelets was over 99%. When the recovered red blood cells were observed under a scanning electron microscope, they showed exactly the same disk shape as before contact with the adsorbent, and when viewed by Perpart's method in terms of osmotic fragility, there was no change at all before and after contact with the adsorbent. In addition, after storing the samples that were in contact with the adsorbent and those that were not in contact with the adsorbent for one week at 4°C, we determined the quantitative determination of adenosine triphosphate and 2,3-diphosphoglycenic acid, and found that there was no difference in the quantitative values. It wasn't accepted.
本発明の方法を実施するための装置としては、第1図に
示すような装置があり、吸着体5を充填し吸着体5をナ
イロンメツシュ4で支持したカラム3に血液バッグ1を
連結して用いる。ヒト全血あるいは赤血球流層等の血液
製剤は中空針7から導管2を通って吸着体5が充填され
たカラム3に導かれ白血球・血小板は吸着体5に付着す
る。そしてこの吸着体5に付着しない赤血球は導管2を
通って血液バッグ1に回収される。また、この装置はオ
ートクレーブで115℃15分処理して無菌化が可能で
ある。この加熱処理で変化をうけるものはmイオン交換
体のジエチルアミノエチルセルロースで他のセルロース
はこの処理で伺ら影響をうけない。As an apparatus for implementing the method of the present invention, there is an apparatus as shown in FIG. 1, in which a blood bag 1 is connected to a column 3 filled with an adsorbent 5 and supported by a nylon mesh 4. used. Blood products such as human whole blood or a red blood cell flow bed are guided from a hollow needle 7 through a conduit 2 to a column 3 filled with an adsorbent 5, and white blood cells and platelets adhere to the adsorbent 5. Red blood cells that do not adhere to the adsorbent 5 are collected into the blood bag 1 through the conduit 2. Additionally, this device can be sterilized by autoclaving at 115° C. for 15 minutes. The only substance that is affected by this heat treatment is diethylaminoethylcellulose, which is an m-ion exchanger, and other celluloses are not affected by this treatment.
以上説明したように、本発明の装置は低摩な吸着剤を用
いて、ヒト全血あるいは赤血球流層から組織抗原を含む
白血球・血小板が完全に除かれた成分輸血用の赤血球の
調製が簡単、かつ経済的にできるようにし、臨床検査用
の純粋な赤血球の調製を容易圧する等の効果を生せしめ
るものである。As explained above, the device of the present invention uses a low-friction adsorbent to easily prepare red blood cells for component transfusion from human whole blood or a red blood cell flow layer, from which white blood cells and platelets containing tissue antigens have been completely removed. It also makes it economical and has the advantage of making it easier to prepare pure red blood cells for clinical tests.
以下、実施例をもって本発明を具体的に説明するO
実施例I
KC−フロックW−50(S)(山陽国策パルプ株式会
社製) 0.5 Fを生理食塩水約5+dK懸濁して、
これを内容積10−の25ミクロンのオープニングを有
するナイロンメツシュ2枚をしいたポリプロピレン製カ
ラムにつめる0加圧して吸着体の容積を2.5−にする
。ヘパリン10 IU/−を含むヒト新鮮全血6.0−
を流す。更に生理食塩水2.0−を流し合計約8−の両
分を得る。所用時間約30分。吸着体を含ま表いカラム
に血液を流しくブランク)同様に処理したもの、および
全血6−に生理食塩水2−を加えたものを同時に血球数
を比較した0
実施例2
KC−フロックW −50(S) 0.5 fを実施例
1と同様にカラムにつめ、赤血球流層(日光)4.0d
K生理食塩水2.0mlを加えて流す0以上実施例1と
同様に処理し、血球カウントした0
実施例3
KCフロックw −50(S) 0.5 tを実施例1
と同様にカラムにつめ兎新鮮血にヘパリン10 IU/
−を加えたもの6.0−を流す0以下実施例1と同様に
処理し、血球をカウントした。Hereinafter, the present invention will be specifically explained with reference to examples.
This is packed into a polypropylene column with an internal volume of 10 and two nylon meshes having an opening of 25 microns, and zero pressure is applied to bring the volume of the adsorbent to 2.5. Human fresh whole blood 6.0- with heparin 10 IU/-
flow. Further, 2.0-liters of physiological saline was poured in to obtain a total of about 8-liters. It takes about 30 minutes. Example 2 KC-Flock W -50 (S) 0.5 f was packed into a column in the same manner as in Example 1, and the red blood cell flow layer (sunlight) was placed at 4.0 d.
Add 2.0 ml of K physiological saline and drain. 0 or more. Treated in the same manner as in Example 1 and counted blood cells.
Similarly, add 10 IU of heparin to fresh rabbit blood in the column.
The same procedure as in Example 1 was carried out, and the blood cells were counted.
10−
実施例4
アビセルpT(101(旭化成工業株式会社製)0.5
fを生理食塩水約5 mAKl!iIi濁して、実施例
1と同様にカラムにつめ吸着体の容積を1.0−とする
。これに赤血球流層(日歩)4.0−に生理食塩水2.
0−を加えて流す。以下実施例1と同様に処理し血球を
カウントした。10- Example 4 Avicel pT (101 (manufactured by Asahi Kasei Corporation) 0.5
f about 5 mAKl in saline! iii) The mixture is turbidly packed into a column in the same manner as in Example 1, and the volume of the adsorbent is set to 1.0-. Add to this the red blood cell flow layer (Nippo) 4.0 - and physiological saline 2.
Add 0- and let it flow. Thereafter, the cells were treated in the same manner as in Example 1, and blood cells were counted.
実施例5
ジエチルアミノエチルセルロース(C1−g )(東京
化成工業株式会社製をC1−型として精製乾燥したもの
)0.5Fを生理食塩水約5−に懸濁し実施例1と同様
にカラムにつめ、赤血球流層(日歩)4.0−に生理食
塩水2.0−を加えて流す。以下実施例1と同様に処理
し血球をカウントした。Example 5 Diethylaminoethylcellulose (C1-g) (purified and dried from Tokyo Chemical Industry Co., Ltd. as C1-type) 0.5F was suspended in about 5-g of physiological saline and packed in a column in the same manner as in Example 1. Physiological saline 2.0- was added to the red blood cell flow bed (Nippo) 4.0- and allowed to flow. Thereafter, the cells were treated in the same manner as in Example 1, and blood cells were counted.
第1図は本発明の装置の1実施例を示す図である。
1・・・血液バッグ、2・・・導管、3・・・カラム、
4・・・ナイロンメツシュ、5・・・吸着体、6・・・
分岐管、7・・・中空針〇
出願人 テルモ株式会社FIG. 1 is a diagram showing one embodiment of the apparatus of the present invention. 1... Blood bag, 2... Conduit, 3... Column,
4... Nylon mesh, 5... Adsorbent, 6...
Branch pipe, 7...Hollow needle〇Applicant Terumo Corporation
Claims (1)
・精製装置において、前記カラムは血液流入口と血液流
出口を有し、カラム内にはセルロース粉末、結晶セルロ
ース、陰イオン交換セルロースのうち少なくともひとつ
からなる吸着体を充欄し、さらに、該吸着体を支持する
メツシュが設けられていることを特徴とする血液の分画
・精製装置。(1) In a blood fractionation/purification device having a column and an adsorbent in the column, the column has a blood inlet and a blood outlet, and the column contains cellulose powder, crystalline cellulose, anion exchange cellulose, etc. 1. A blood fractionation and purification device, characterized in that it is filled with an adsorbent made of at least one of the above, and is further provided with a mesh that supports the adsorbent.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP58204243A JPS5995051A (en) | 1983-10-31 | 1983-10-31 | Apparatus for fractionating and purifying blood |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP58204243A JPS5995051A (en) | 1983-10-31 | 1983-10-31 | Apparatus for fractionating and purifying blood |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS5995051A true JPS5995051A (en) | 1984-05-31 |
JPH0326170B2 JPH0326170B2 (en) | 1991-04-10 |
Family
ID=16487216
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP58204243A Granted JPS5995051A (en) | 1983-10-31 | 1983-10-31 | Apparatus for fractionating and purifying blood |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS5995051A (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2014126014A1 (en) | 2013-02-12 | 2014-08-21 | 東レ株式会社 | Blood purification column |
-
1983
- 1983-10-31 JP JP58204243A patent/JPS5995051A/en active Granted
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2014126014A1 (en) | 2013-02-12 | 2014-08-21 | 東レ株式会社 | Blood purification column |
KR20150118085A (en) | 2013-02-12 | 2015-10-21 | 도레이 카부시키가이샤 | Blood purification column |
US10213543B2 (en) | 2013-02-12 | 2019-02-26 | Toray Industries, Inc. | Blood purification column |
Also Published As
Publication number | Publication date |
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JPH0326170B2 (en) | 1991-04-10 |
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