JPS5854127B2 - Filter for leukocyte separation - Google Patents

Description

【発明の詳細な説明】 この発明は血液、体液等の血球浮遊液から白血球を選択
的に分離するためのフィルターに関するものである。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a filter for selectively separating white blood cells from a blood cell suspension such as blood or body fluid.

さらに詳しく述べると、血液、骨髄液、リンパ液などの
血球を含む体液、およびこれらの液に何らかの処理を加
えて得た血球を含む液体、たとえばデキストランやヒド
ロキシエチルスターチなどの赤血球凝集剤を加えて得た
赤血球の大部分が除去された血液、蒸留水や０．２％Ｎ
ａＣ１水溶液などの低張溶液と接触させて赤血球を破壊
した血液、などの少なくとも白血球とそれ以外の物質を
含む液体から、白血球だけを分離する、さらには白血球
の各種類にも分離できる、白血球分離用フィルターに関
するものである。To be more specific, body fluids containing blood cells such as blood, bone marrow fluid, and lymph, and fluids containing blood cells obtained by subjecting these fluids to some kind of treatment, such as those obtained by adding red blood cell agglutinants such as dextran or hydroxyethyl starch. Blood from which most of the red blood cells have been removed, distilled water or 0.2% N
Leukocyte separation, which separates only white blood cells, or even separates each type of white blood cell, from a liquid containing at least white blood cells and other substances, such as blood whose red blood cells have been destroyed by contact with a hypotonic solution such as aC1 aqueous solution. This relates to filters for

近年、赤血球、白血球、血小板等を血液から分離し、種
々の疾患の治療のために輸血することが行われるように
なってきた。In recent years, it has become common to separate red blood cells, white blood cells, platelets, etc. from blood and transfuse them for the treatment of various diseases.

このような血液成分輸血は、従来の全血輸血とは異なり
、不必要なものは除去して必要な成分だけを濃縮して輸
血できるという大きな利点をもっており、各地の病院で
治療に用いられている。Unlike conventional whole blood transfusions, this type of blood component transfusion has the great advantage of removing unnecessary components and concentrating only the necessary components for transfusion, and is used for treatment at hospitals all over the country. There is.

特に白血球の輸血は、白血球が異常に低下し、細菌など
により発熱した患者の治療や免疫療法に広く各地の病院
で行われている。In particular, transfusions of white blood cells are widely used in hospitals across the country for the treatment and immunotherapy of patients with abnormally low white blood cell counts and fever caused by bacteria.

このような目的に使用可能な従来の血球分離技術として
は、血球の比重差を利用した重力遠心分離方法、血球の
粘着、付着等の作用を利用した捕捉材利用の方法、赤血
球凝集剤を用いた白血球分離方法等がある。Conventional blood cell separation techniques that can be used for this purpose include gravity centrifugation that uses the difference in specific gravity of blood cells, methods that use capture materials that take advantage of the adhesion and adhesion of blood cells, and methods that use red blood cell agglutination agents. There are various leukocyte separation methods.

血球の分離方法としてよく用いられているものは、赤血
球、白血球、血漿等その成分毎に異なる比重を利用した
、分離したい血球と同じ比重の液体に血液を重層して遠
心分離操作を行う密度勾配遠心分離法、デキストラン、
ヒドロキシエチルスターチ等の赤血球凝集剤によって赤
血球を沈降させ、白血球および血小板の浮遊液を得る方
法、白血球のうち顆粒球を捕捉する物質を入れたカラム
に血液を流し、捕捉物質に捕捉された顆粒球を回収して
顆粒球を得る方法、などである。A commonly used method for separating blood cells is density gradient, which utilizes the different specific gravity of each component such as red blood cells, white blood cells, and plasma, and involves layering blood in a liquid with the same specific gravity as the blood cells to be separated and performing a centrifugal separation operation. centrifugation, dextran,
A method in which red blood cells are precipitated with a red blood cell aggregating agent such as hydroxyethyl starch to obtain a suspension of white blood cells and platelets.Blood is poured into a column containing a substance that captures granulocytes among white blood cells, and the granulocytes are captured by the capture substance. A method for obtaining granulocytes by collecting granulocytes.

しかし、これらの方法で得られる血球は純度が低く、供
給した総画血球数に対して分離された白血球の収率が低
い。However, the blood cells obtained by these methods have low purity, and the yield of separated white blood cells is low relative to the total number of blood cells supplied.

すなわち分離効率が低い。また白血球を分離するための
装置も大型かつ高価になったり、操作が煩雑であったり
し、大病院では使えても中小病院で簡単に使えるもので
はなかった。In other words, the separation efficiency is low. Additionally, devices for separating white blood cells are large, expensive, and complicated to operate, and although they can be used in large hospitals, they cannot be easily used in small and medium-sized hospitals.

すなわち、密度勾配遠心分離方法や連続遠心分離方法等
、遠心操作を行う方法は、通常必要とされる血液量を処
理するためには大型の遠心分離器を必要とする。That is, methods that involve centrifugation, such as density gradient centrifugation and continuous centrifugation, typically require large centrifuges to process the required amount of blood.

そして分離された白血球中に含まれる不必要な成分は、
さらに数回の洗浄操作、すなわち遠心分離操作を必要と
する。The unnecessary components contained in the separated white blood cells are
Several additional washing operations, ie, centrifugation operations, are required.

これらの操作を行う際、白血球の二部が破壊され、白血
球の収率が悪くなるとともに、一連の操作には熟練を要
し、非常に煩雑な操作である。When performing these operations, two parts of the leukocytes are destroyed, resulting in a poor yield of leukocytes, and the series of operations requires skill and is extremely complicated.

赤血球凝集剤を用いる方法は、比較的簡便な操作により
白血球および血小板の血漿浮遊液を得ることができるが
、白血球、血小板とともに白血球の数倍から数十倍量の
赤血球および多量の蛋白質等が混じってしまい、これだ
けで白血球、血小板分離ができたとは言えない。The method using a hemagglutinating agent allows a plasma suspension of white blood cells and platelets to be obtained through a relatively simple operation, but the white blood cells and platelets are mixed with red blood cells, which are several times to several tens of times the amount of white blood cells, and a large amount of protein. Therefore, it cannot be said that leukocytes and platelets could be separated using this method alone.

また、白血球の収率も悪い。さらに白血球の純度を上げ
るためには、上述した遠心分離方法に頼らなければなら
なくなってしまう。Furthermore, the yield of white blood cells is also poor. Furthermore, in order to increase the purity of leukocytes, it becomes necessary to rely on the above-mentioned centrifugation method.

顆粒球を捕捉する物質を用いる方法は、白血球のうち顆
粒球の選択分離が可能であるが、特別な装置が必要であ
り、操作が煩雑で顆粒球の収率も悪く、患者に輸血する
ための適正濃度にするためにはやはり前述の遠心分離が
必要である。Methods using substances that capture granulocytes enable the selective separation of granulocytes among white blood cells, but require special equipment, are complicated to operate, have a poor yield of granulocytes, and require blood transfusion to the patient. The above-mentioned centrifugation is still necessary to obtain the appropriate concentration.

そこで我々は、これらの問題点を解決し、さらに高い収
率で純度の高い白血球が簡単な操作で得られる白血球分
離フィルターを提供するとともに、白血球の各成分にも
分離可能な白血球分離フィルターを提供することを目的
に鋭意研究した。Therefore, we have solved these problems by providing a leukocyte separation filter that can obtain high-yield and highly pure white blood cells with simple operations, and also provide a leukocyte separation filter that can also separate each component of white blood cells. I conducted extensive research with the aim of doing so.

その結果、平均直径が１０μ肌より大きな平均直径の繊
維状物質の集合体で構成したフィルターでは、血球のう
ち白血球の一部程度しか分離できないが、平均直径１０
μ扉以下の繊維状物質を使用すれば白血球のほとんどを
純度良くフィルターに捕捉できることを、我々は新たに
見出した。As a result, a filter composed of aggregates of fibrous substances with an average diameter larger than the average diameter of 10μ skin can separate only a portion of white blood cells among blood cells;
We have newly discovered that most of the white blood cells can be captured in a filter with high purity by using a fibrous material with a size smaller than the μ door.

しかし、平均直径１０μ扉以下の繊維状物質だけで構成
したフィルターを使用した場合、白血球のほとんどを純
度良く捕捉することは可能となったが、白血球を回収す
る操作によって得られる白血球が、小量の回収液ではや
や少なかった。However, when using a filter made only of fibrous material with an average diameter of 10μ or less, it became possible to capture most of the white blood cells with good purity, but the white blood cells obtained by the operation of collecting the white blood cells were small. It was slightly less in the recovered liquid.

すなわち回収率がやや悪く、６０％程度であった。That is, the recovery rate was rather poor, at about 60%.

そこでさらに研究を進めた結果、平均直径が１０μ扉以
下の繊維状物質に平均直径が１０μ肌より大きい繊維状
物質を混ぜてやることにより、回収率を高めることに成
功した。As a result of further research, they succeeded in increasing the recovery rate by mixing a fibrous material with an average diameter of 10 microns or less with a fibrous material with an average diameter of 10 microns or less.

すなわち、白血球の捕捉率は平均直径が１０μ扉以下の
繊維状物質だけで構成したフィルターと同等かそれより
やや低い程度であったが、白血球の回収率は平均直径が
１０μ胤以下の繊維状物質だけで構成したフィルターよ
り高く、７０〜８０％の回収率で白血球を回収できる。In other words, the capture rate of white blood cells was comparable to or slightly lower than that of a filter made only of fibrous material with an average diameter of 10 μm or less; White blood cells can be recovered at a recovery rate of 70 to 80%, which is higher than a filter constructed with only a single filter.

さらに、平均直径が１０μｍ以下の繊維状物質の材質、
および平均直径が１０μｍ以下の繊維状物質と１０μ扉
より大きい平均直径の繊維状物質との混合比や、平均直
径の選び方等により、白血球のうち単球と顆粒球だけを
純度良く捕捉できることをも見出し、本発明を完成させ
た。Furthermore, the material of the fibrous substance has an average diameter of 10 μm or less,
It has also been shown that only monocytes and granulocytes among white blood cells can be captured with high purity by adjusting the mixing ratio of fibrous substances with an average diameter of 10 μm or less and fibrous substances with an average diameter larger than 10 μm, and by selecting the average diameter. The present invention has been completed.

すなわち本発明は、繊維状物質の集合体を主要部とする
血球分離用フィルターであって、平均直径１μ扉以上１
０μ扉以下の繊維状物質一種類以上と、１０μ扉より大
きい平均直径の繊維状物質一種類以上との混合集合体を
フィルターの主要部とすることを特徴とした白血球分離
用のフィルターである。That is, the present invention provides a filter for separating blood cells, which has an aggregate of fibrous substances as its main part, and which has an average diameter of 1μ or more.
This filter for leukocyte separation is characterized in that the main part of the filter is a mixed aggregate of one or more types of fibrous substances with an average diameter of 0μ or less and one or more types of fibrous substances with an average diameter larger than 10μ.

この発明の血球分離用フィルターは、例えば第１図のよ
うに構成される。The blood cell separation filter of the present invention is constructed as shown in FIG. 1, for example.

密封容器１には血球浮遊液の導入口２および排出口３が
取り付けられていて、密封容器１内に平均直径１μ流以
上１０μ扉以下の繊維状物質と１０μ乳より大きい平均
直径の繊維状物質との混合集合体４が詰め込まれている
。The sealed container 1 is equipped with an inlet 2 and an outlet 3 for a blood cell suspension, and a fibrous substance with an average diameter of 1 μm or more and a flow of 10 μm or less and a fibrous material with an average diameter larger than 10 μm flow into the sealed container 1. A mixed aggregate 4 is packed.

網５は、繊維状物質の混合集合体４が血球浮遊液の流れ
によって排出口３の方向に流出するのを防ぐもので、な
くても良いが、あった方がフィルターの目詰まりを防ぐ
うえで有効である。The net 5 prevents the mixed aggregate 4 of fibrous substances from flowing out in the direction of the outlet 3 due to the flow of blood cell suspension, and although it is not necessary, it is better to prevent the filter from clogging. is valid.

ここで言う繊維状物質とは、繊維状物質の平均直径に対
して繊維状物質の長さが非常に長いものを言う。The fibrous material referred to here refers to a fibrous material whose length is extremely long relative to its average diameter.

材質は合成繊維、金属繊維、天然繊維等繊維状の形を保
てるものであれば何でも良い。The material may be anything that can maintain its fibrous shape, such as synthetic fibers, metal fibers, and natural fibers.

繊維状物質の平均直径とは、繊維の種類によって、その
横断面が円ではなく、星形、楕円形等さまざまなものが
あるが、その断面を円形に換算したときの直径を言い、
種々の直径の繊維状物質の平均直径という意味ではない
。The average diameter of a fibrous substance is the diameter when the cross section is converted into a circle, although the cross section may be star-shaped, oval, etc., rather than circular depending on the type of fiber.
It does not mean the average diameter of fibrous materials of various diameters.

また繊維状物質の混合集合体とは、異った平均直径の繊
維状物質同士が均一に分散している状態をいい、極端な
片寄りがあってはならない。A mixed aggregate of fibrous materials refers to a state in which fibrous materials with different average diameters are uniformly dispersed, and there must be no extreme deviation.

また、繊維状物質同士は、より合わされたりしないで、
綿状に分散していることが望ましい。In addition, fibrous substances should not be twisted together,
It is desirable that it is dispersed in a flocculent form.

容器１への繊維状物質の混合集合体４の詰め方も、均一
に分散させた状態にするのが望ましいが、容器１の一部
に空間を作ったり、充填密度を変えたものを組み合わせ
ても差し支えない。Regarding the method of filling the mixed aggregate 4 of fibrous substances into the container 1, it is desirable to make it uniformly dispersed, but it is also possible to create a space in a part of the container 1 or to combine materials with different packing densities. There is no problem.

異なった平均直径の繊維状物質の組み合わせ方の例を以
下に示す。Examples of how to combine fibrous materials of different average diameters are shown below.

白血球のほとんどを捕捉し、赤血球、血小板、血漿をｒ
液として得ようとする場合には、例えば平均直径１μ扉
以上１０μ扉以下の繊維状物質に対し、その平均直径の
１．５倍から８倍程度の平均直径の繊維状物質を２０〜
８０重量％混合してやる。Captures most of the white blood cells and removes red blood cells, platelets, and plasma.
If you want to obtain it as a liquid, for example, for a fibrous material with an average diameter of 1 μm or more and 10 μm or less, 20 to 20 times the average diameter of the fibrous material is about 1.5 times to 8 times the average diameter.
Mix 80% by weight.

その混合された繊維状物質がどちらも平均直径１０μ扉
以下の場合は、血球浮遊液の流れによってフィルターを
構成している繊維状物質同士がつぶされ、目詰まりを起
こし易くなったり、白血球の回収率が悪くなったりする
ので、平均直径が１０μ扉より大きい繊維状物質をさら
に追加してやり、目詰まりや白血球の回収率が低下する
のを防いでやる。If both of the mixed fibrous substances have an average diameter of 10μ or less, the fibrous substances that make up the filter will be crushed by the flow of the blood cell suspension, making it easier to clog or collecting white blood cells. Since the rate may deteriorate, a fibrous material with an average diameter larger than 10 μm is added to prevent clogging and a decrease in the leukocyte recovery rate.

このとき、平均直径が１０μｍより大きい繊維状物質を
２種類以上混ぜてやっても同様の結果が得られる。At this time, similar results can be obtained even if two or more types of fibrous substances having an average diameter of more than 10 μm are mixed.

平均直径１０μｍより大きい繊維状物質だけでは、どの
ような組み合わせ方をしても、白血球のすべてを捕捉し
、赤血球、血小板、血漿をスムーズに沢過できるような
間隙を作ることはできなかった。No matter how the fibrous material, which has an average diameter of more than 10 μm, is combined, it is not possible to capture all the white blood cells and create a gap through which red blood cells, platelets, and plasma can smoothly pass through.

フィルターに接触させる血球浮遊液の流速等により組合
せは多少変わるが、この発明では平均直径１０μ肌以下
の繊維状物質を適当に混ぜてやることによってこれを可
能とした。Although the combination varies somewhat depending on the flow rate of the blood cell suspension brought into contact with the filter, this invention has been made possible by appropriately mixing fibrous substances with an average diameter of 10 microns or less.

さらに、平均直径１０μ肌以下の繊維状物質だけで構成
したフィルターは、捕捉した白血球を回収する際、その
回収率があまり良くなかったのに対し、この発明のフィ
ルターは捕捉された白血球の回収率が高いというメリッ
トもある。Furthermore, a filter composed only of fibrous material with an average diameter of 10 μm or less had a poor recovery rate when collecting captured white blood cells, whereas the filter of this invention had a low recovery rate of captured white blood cells. It also has the advantage of being high.

白血球のうち、単球および顆粒球だけを捕捉し、リンパ
球および赤血球、血小板、血漿をＦ液として得ようとす
る場合は、平均直径１０μ流以下の繊維状物質と、１０
μ招より大きい平均直径の繊維状物質同士の作る空間が
、リンパ球、赤血球等を捕捉しない大きさに、繊維状物
質の組み合わせ方を調節してやる。If you want to capture only monocytes and granulocytes among white blood cells and obtain lymphocytes, red blood cells, platelets, and plasma as F fluid, fibrous material with an average diameter of 10μ or less and 10
The way the fibrous materials are combined is adjusted so that the space created by the fibrous materials, which have an average diameter larger than μ, does not trap lymphocytes, red blood cells, etc.

この場合は、さらに、使用する繊維状物質の材質として
、ポリアミド、ポリアクリロニトリル系、再生セルロー
ス、ポリエステルのうち一種類以上の繊維状物質を使用
し、単球および顆粒球を捕捉してやる。In this case, the fibrous material used is one or more of polyamide, polyacrylonitrile, regenerated cellulose, and polyester to trap monocytes and granulocytes.

平均直径が１０μ扉より大きい繊維状物質だけでは、ど
のような材質を用いても、単球、顆粒球だけを純度良く
高い収率で捕捉することはできなかったが、平均直径１
０μ肌以下の繊維状物質を混ぜてやることにより、単球
、顆粒球の捕捉率が上がることが見出された。No matter what kind of material is used, it was not possible to capture only monocytes and granulocytes with high purity and high yield using only fibrous substances with an average diameter larger than 10 μm.
It has been found that the capture rate of monocytes and granulocytes can be increased by mixing fibrous substances with a particle size of 0μ or less.

上記これらのフィルターの組み合わせにより、リンパ球
とそれ以外の白血球の選択捕捉が可能であることは言う
までもない。It goes without saying that the combination of these filters allows selective capture of lymphocytes and other white blood cells.

この発明の白血球分離用フィルターは、例えば第２図の
ような装置により使用される。The leukocyte separation filter of the present invention is used, for example, with an apparatus as shown in FIG.

血球浮遊液６は容器７に溜められ、ポンプ８により白血
球分離フィルター９に送られる。The blood cell suspension 6 is collected in a container 7 and sent to a leukocyte separation filter 9 by a pump 8.

このとき弁１０は開かれ、弁１１．１３は閉じられてい
る。At this time, valve 10 is open and valve 11.13 is closed.

白血球分離フィルター９に送られた血球浮遊液６は、白
血球分離フィルター９で沢過され、白血球はフィルター
９内に捕捉される。The blood cell suspension 6 sent to the leukocyte separation filter 9 is thoroughly filtered by the leukocyte separation filter 9, and the leukocytes are captured within the filter 9.

他の成分は弁１５または１６を通って容器１２または２
０に送られる。Other ingredients pass through valve 15 or 16 to container 12 or 2.
Sent to 0.

血球浮遊液６がすべて容器７外に送り出された時点で弁
１０が閉じ、弁１１または１７が開かれる。When all of the blood cell suspension 6 has been sent out of the container 7, the valve 10 is closed and the valve 11 or 17 is opened.

すると容器１３または１８内の洗浄液１４または１９が
血球浮遊液６と同じ回路を通って白血球分離フィルター
９内に送られ、白血球分離フィルター９で捕捉された白
血球以外の成分を洗い流す。Then, the washing liquid 14 or 19 in the container 13 or 18 is sent into the leukocyte separation filter 9 through the same circuit as the blood cell suspension 6, and the components other than leukocytes captured by the leukocyte separation filter 9 are washed away.

この後、適当な方法で白血球分離フィルター９内の白血
球を回収してやれば純粋な白血球が得られる。Thereafter, by collecting the leukocytes in the leukocyte separation filter 9 using an appropriate method, pure leukocytes can be obtained.

ここで、ポンプ８は血球に悪影響を与えないものであれ
ばどんなタイプを使用しても良いが、市販の血球ポンプ
などが使い易い。Here, any type of pump 8 may be used as long as it does not adversely affect blood cells, but commercially available blood cell pumps are easy to use.

白血球分離フィルター９は、平均直径１０μ扉以下の繊
維状物質と１０μ扉より大きい平均直径の繊維状物質と
の混合集合体を主要部としており、分離したい白血球の
種類によって、その繊維状物質の平均直径、混合割合、
材質等が任意に選ばれる。The leukocyte separation filter 9 mainly consists of a mixed aggregate of fibrous substances with an average diameter of 10 μm or less and fibrous substances with an average diameter larger than 10 μm. diameter, mixing ratio,
The material etc. can be selected arbitrarily.

洗浄液は、生理食塩水、リンゲル液等、白血球に悪影響
を与えないものであれば何でも良く、白血球分離フィル
ターに残存する不必要成分を洗い流しでや−る目的で使
用する液体である。The washing liquid may be anything such as physiological saline or Ringer's solution as long as it does not have an adverse effect on leukocytes, and is a liquid used for the purpose of washing away unnecessary components remaining in the leukocyte separation filter.

以上のようにして得られる白血球は、純度、収率ともに
良く、操作も短時間かつ非常に簡単で、大型の装置も全
く不要である。The leukocytes obtained in the above manner have good purity and yield, are very simple to operate in a short time, and do not require any large equipment.

実施例 １ 健康人の血液（赤血球数５００万／ｌｎ４、白血球数７
０００７ｍ７１！、血小板数２５万／ｍ４．総蛋白濃度
７ ｙ／ｄｌ ） １００ｒｒＬｌにヘパリンを１００
０単位採血時に加え、この血液を血球浮遊液として以下
の実験に使用した。Example 1 Blood of a healthy person (red blood cell count 5 million/ln4, white blood cell count 7
0007m71! , platelet count 250,000/m4. Total protein concentration 7 y/dl) 100rrL of heparin
0 units were added at the time of blood collection, and this blood was used as a blood cell suspension in the following experiments.

この血球浮遊液を第２図に示す実験回路を用いて処Ｗし
た。This blood cell suspension was treated using the experimental circuit shown in FIG.

白血球分離フィルター９としては、平均直径３．５μ扉
のポリアクリロニトリル繊維２ｇｒと、平均直径２３μ
肌のポリアクリロニトリル繊維１．５ｇｒをよく混合し
て、直径１８１１ｔ？＋！、長さ１００ｍｍの容器に詰
めたものを使用した。The leukocyte separation filter 9 is made of 2gr polyacrylonitrile fiber with an average diameter of 3.5μ and an average diameter of 23μ.
Mix well 1.5g of skin polyacrylonitrile fiber and make a diameter of 1811t? +! A container with a length of 100 mm was used.

ここで用いた繊維の長さは４０〜８０１１ｔ１ｒＬであ
った。The length of the fibers used here was 40 to 8011t1rL.

前述した血球浮遊液６を容器７に入れ、ポンプ８により
白血球分離フィルター９に５ ｒｔｔｌ／ｍｍで送った
。The aforementioned blood cell suspension 6 was placed in a container 7 and sent to a leukocyte separation filter 9 at a rate of 5 rttl/mm by a pump 8.

このとき弁１０，１５は開き、弁１１，１６゜１７は閉
じた状態である。At this time, valves 10 and 15 are open, and valves 11, 16 and 17 are closed.

血球浮遊液６が容器７から完全に排出されたところで弁
１０を閉じ、弁１１を開いて洗浄液１４をポンプ８によ
り１０ｍ１／ｍｉｎの流量でフィルター９に送った。When the blood cell suspension 6 was completely discharged from the container 7, the valve 10 was closed, the valve 11 was opened, and the washing liquid 14 was sent to the filter 9 by the pump 8 at a flow rate of 10 ml/min.

２００ｍ１流れた所で容器１２に溜まった沢液を検査し
たところ、この実験に供した血球浮遊液の総量に対して
、蛋白質は９９．５％、赤血球は９９．７％、血小板は
９４％回収されたのに対し、白血球は１０％であった。When we inspected the solution collected in container 12 after 200 m1 flowed, we found that 99.5% of protein, 99.7% of red blood cells, and 94% of platelets were recovered from the total amount of blood cell suspension used in this experiment. white blood cell count was 10%.

すなわち、白血球分離フィルター９に９０％の白血球が
捕捉された。That is, 90% of the leukocytes were captured in the leukocyte separation filter 9.

この後、弁１１．１５を閉じ、弁１７を開き、容器１８
内の回収液１９をポンプ８により白血球分離フィルター
９に５ｍｌ／ｒｎｉ１１．で送り、白血球分離フィルタ
ー９に振動を与えながら容器２０に白血球浮遊液を１０
０７７１１得た。After this, valve 11.15 is closed, valve 17 is opened and container 18
The collected liquid 19 is transferred to the leukocyte separation filter 9 by the pump 8 at 5 ml/rni11. While applying vibration to the leukocyte separation filter 9, 10% of the leukocyte suspension was added to the container 20.
I got 07711.

回収液１９としてはＡＣＤ加アルブミン溶液を用いた。As the recovery liquid 19, an ACD-added albumin solution was used.

容器２０に得られた白血球浮遊液を検査したところ、こ
の実験に供された血球浮遊液の総量に対して、白血球が
７５％という高い回収率で得られていた。When the leukocyte suspension obtained in the container 20 was examined, it was found that leukocytes were obtained at a high recovery rate of 75% based on the total amount of the blood cell suspension used in this experiment.

比較例 １ 実施例１と同じ実験回路を使用し、白血球分離フィルタ
ー９として、平均直径１３μ扉のポリアミド繊維２ｇｒ
、と、平均直径２０μｍのポリアミド繊維２ｇｒをよ（
混ぜて直径１８朋、長さ１００７Ｗ７にの容器に詰めた
ものを使用した以外は、実施例１と同じ条件で血液を処
理した。Comparative Example 1 Using the same experimental circuit as in Example 1, a 2gr polyamide fiber with an average diameter of 13μ was used as the leukocyte separation filter 9.
, and 2 gr of polyamide fibers with an average diameter of 20 μm (
Blood was treated under the same conditions as in Example 1, except that the mixture was mixed and packed in a container with a diameter of 18 mm and a length of 1007 W7.

その結果得られた沢液を調べたところ、蛋白質は９９．
５％、血小板は９４％、赤血球は９９．７％回収された
が、白血球も４５％含まれていた。When the resulting saliva was examined, the protein content was 99.
5%, platelets were recovered at 94%, red blood cells at 99.7%, and white blood cells at 45%.

すなわち、白血球分離フィルター９内の繊維の密度は実
施例１よりあがったのにもかかわらず、白血球は５５％
しか捕捉されなかった。In other words, although the density of the fibers in the leukocyte separation filter 9 was higher than in Example 1, the leukocyte concentration was 55%.
only was captured.

また捕捉された白血球の内訳は、リンパ球３０％に対し
単球、顆粒球が７０％であり、リンパ球もかなり高い濃
度で捕捉されていた。Furthermore, the captured white blood cells were 30% lymphocytes and 70% monocytes and granulocytes, and lymphocytes were also captured at a fairly high concentration.

また、実施例１と同様に、フィルターに捕捉された白血
球を回収したところ、この実験に供された血球浮遊液の
総量に対して、白血球は４５％しか回収されなかった。Furthermore, in the same manner as in Example 1, when the leukocytes captured on the filter were recovered, only 45% of the leukocytes were recovered with respect to the total amount of the blood cell suspension used in this experiment.

比較例 ２ 実施例１と同じ実験回路を使用し、白血球分離フィルタ
ー９として、平均直径３．５μ肌のポリアクリロニトリ
ル繊維３．５ｇｒ、だげを直径１８ｍｒＩＬ、長さｉｏ
ｏｍｍの容器に詰めたものを使用した以外は、実施例１
と同じ条件で血液を処理した。Comparative Example 2 The same experimental circuit as in Example 1 was used, and as the leukocyte separation filter 9, 3.5g of polyacrylonitrile fiber with an average diameter of 3.5μ, a diameter of 18mIL, and a length of IO were used.
Example 1 except that the one packed in an omm container was used.
Blood was processed under the same conditions.

その結果得られたＰ液を調べたところ、蛋白質は９９．
５％、赤血球は９９，６％、血小板は９４％回収された
のに対し、白血球は２％であった。When the resulting P solution was examined, the protein content was 99.
5%, 99.6% red blood cells, 94% platelets, and 2% white blood cells.

すなわち、白血球分離フィルター９に９８％もの白血球
が捕捉された。That is, 98% of the leukocytes were captured in the leukocyte separation filter 9.

しかし、実施例１と同様に、捕捉された白血球を回収し
たところ、この実験に供された血球浮遊液の総量に対し
て、白血球は５５％しか回収できなかった。However, when the captured leukocytes were recovered in the same manner as in Example 1, only 55% of the leukocytes could be recovered with respect to the total amount of the blood cell suspension used in this experiment.

実施例 ２ 実施例１と同じ実験回路を使用し、白血球分離フィルタ
ー９として、平均直径７μ爪のポリアミド繊維１．５ｇ
ｒ、と、平均直径１５μ扉のポリアミド繊維１．５ｇｒ
、とを、直径１８朋、長さ１００ｍｍの容器に詰めたも
のを使用した以外は、実施例１と同じ条件で血液を処理
した。Example 2 Using the same experimental circuit as in Example 1, 1.5 g of polyamide fiber with an average diameter of 7 μm was used as the leukocyte separation filter 9.
r, and polyamide fiber 1.5gr with an average diameter of 15μ door
Blood was treated under the same conditions as in Example 1, except that a container with a diameter of 18 mm and a length of 100 mm was used.

その結果得られたｆ液を検査したところ、蛋白質は９９
．５％、血小板は９２％、赤血球は９９．６％回収され
、白血球も５０％回収された。When the resulting f solution was examined, the protein was found to be 99%.
．． 5%, platelets were recovered by 92%, red blood cells by 99.6%, and white blood cells by 50%.

しかし、捕捉された白血球の内訳は、単球、顆粒球が９
７％、リンパ球が３％であり、高い純度で半球顆粒球が
捕捉された。However, the breakdown of captured white blood cells was that monocytes and granulocytes were 9
7%, lymphocytes 3%, and hemispheric granulocytes were captured with high purity.

実施例 ３ 実施例１と同じ実験回路を使用し、白血球分離フィルタ
ー９として、平均直径８μ椛のポリアクリロニトリル繊
維１．５ｇｒ、と、平均直径１５μ流のポリアクリロニ
トリル繊維１．５ｇｒ、とを良く混合したものを使用し
た以外は、実施例１と同じ条件で血液を処理した。Example 3 Using the same experimental circuit as in Example 1, as the leukocyte separation filter 9, 1.5 gr of polyacrylonitrile fibers with an average diameter of 8 μm and 1.5 gr of polyacrylonitrile fibers with an average diameter of 15 μm were thoroughly mixed. Blood was treated under the same conditions as in Example 1, except that blood was used.

その結果得られた沢液を検査したところ、蛋白質は９９
．５％、赤血球は９９．５％、血小板は９４％回収され
、白血球も４８％回収された。When the resulting saliva was tested, the protein content was 99%.
．． 5%, red blood cells were recovered by 99.5%, platelets by 94%, and white blood cells by 48%.

しかし捕捉された白血球の内訳は、半球、顆粒球が９６
％、リンパ球が４％であり、高い純度で単球、顆粒球が
捕捉された。However, the breakdown of captured white blood cells was 96 hemispheres and granulocytes.
%, lymphocytes were 4%, and monocytes and granulocytes were captured with high purity.

実施例 ４ 実施例１と同じ実験回路を使用し、白血球分離フィルタ
ー９として、平均直径７．５μ流の再生セルロース繊維
１．５ｇｒ、と、平均直径１３μ流の再生セルロース繊
維１．８ ｇｒ、とを良く混合したものを使用した以外
は、実施例１と同じ条件で血液を処理した。Example 4 The same experimental circuit as in Example 1 was used, and as the leukocyte separation filter 9, 1.5 gr of regenerated cellulose fiber with an average diameter of 7.5 μ flow, and 1.8 gr of regenerated cellulose fiber with an average diameter of 13 μ flow. Blood was treated under the same conditions as in Example 1, except that a well-mixed product was used.

その結果得られた汁液を検査したところ、蛋白質は９９
．７％、赤血球は９９．７％、血小板は９４％回収され
、白血球も５２％回収された。When the resulting juice was examined, the protein content was 99.
．． 7%, 99.7% of red blood cells, 94% of platelets, and 52% of white blood cells.

しかし捕捉された白血球の内訳は、単球、顆粒球が９５
％、リンパ球が５％であり、高い純度で単球、顆粒球が
捕捉された。However, the breakdown of captured white blood cells was 95 monocytes and granulocytes.
%, lymphocytes were 5%, and monocytes and granulocytes were captured with high purity.

実施例 ５ 実施例１と同じ条数回路を使用し、白血球分離フィルタ
ー９として、平均直径６．５μ扉のポリエステル繊維１
．８ｇｒ、と、平均直径１８μ肌のポリエステル繊維１
．６ｇｒ、とを良く混合したものを使用した以外は、実
施例１と同じ条件で血液を処理した。Example 5 Using the same thread number circuit as in Example 1, a polyester fiber 1 with an average diameter of 6.5μ was used as the white blood cell separation filter 9.
．． 8gr, average diameter 18μ skin polyester fiber 1
．． Blood was treated under the same conditions as in Example 1, except that a well-mixed mixture of 6gr and 6gr was used.

その結果得られたｐ液を検査したところ、蛋白質は９９
．７％、赤血球は９９．５％、血小板は９３％回収され
、白血球も５３％回収された。When the resulting p solution was tested, the protein content was 99%.
．． 7%, red blood cells were recovered by 99.5%, platelets by 93%, and white blood cells by 53%.

しかし捕捉された白血球の内訳は、単球、顆粒球９７％
、リンパ球が３％であり、高い純度で半球、顆粒球が捕
捉された。However, the breakdown of captured white blood cells was 97% monocytes and granulocytes.
, lymphocytes were 3%, and hemispheres and granulocytes were captured with high purity.

以上述べたように、この発明の白血球分離フィルターを
使用することにより、従来のフィルターでは不可能であ
った高純度の白血球を７０〜８０％という高い回収率で
簡単に得られるようになった。As described above, by using the leukocyte separation filter of the present invention, it has become possible to easily obtain highly purified leukocytes with a high recovery rate of 70 to 80%, which was impossible with conventional filters.

また、高い純度高い収率で、単球、顆粒球を選択的に捕
捉することもできた。It was also possible to selectively capture monocytes and granulocytes with high purity and high yield.

【図面の簡単な説明】[Brief explanation of drawings]

第１図は本発明による白血球分離用のフィルターの例を
示す模式図であり、第２図は本発明の白血球分離用のフ
ィルターを使用した回路の例を示す模式図である。 １．７，１２．１３，１８，２０〜容器、２〜導入口、
３〜排出口、４−４維状物質の混合集合体、５〜網、８
〜ポンプ、９〜白血球分離用フィルター。FIG. 1 is a schematic diagram showing an example of a filter for separating leukocytes according to the present invention, and FIG. 2 is a schematic diagram showing an example of a circuit using the filter for separating leukocytes of the present invention. 1.7, 12.13, 18, 20 ~ container, 2 ~ inlet,
3 - outlet, 4-4 mixed aggregate of fibrous substances, 5 - net, 8
~Pump, 9~White blood cell separation filter.

Claims (1)

【特許請求の範囲】 １ 繊維状物質の集合体を主要部とする血球分離用フィ
ルターであって、平均直径ｌμ扉以上１０μ乳以下の繊
維状物質一種類以上と、１０μ肌より大きい平均直径の
繊維状物質一種類以上との混合集合体をフィルターの主
要部とすることを特徴とした白血球分離用のフィルター
。 ２ 平均直径１μ肌以上１０μ扉以下の繊維状物質とし
て、ポリアミド、ポリアクリロニトリル系、再生セルロ
ース、ポリエステルのうちの少なくとも１種類の繊維状
物質を使用することを特徴とする特許請求の範囲第１項
記載の白血球分離用のフィルター。[Scope of Claims] 1. A blood cell separation filter mainly composed of aggregates of fibrous substances, which comprises one or more types of fibrous substances with an average diameter of lμ or more and 10μ or less and an average diameter larger than 10μ. A filter for separating leukocytes, characterized in that the main part of the filter is a mixed aggregate with one or more types of fibrous substances. 2. Claim 1, characterized in that at least one type of fibrous material selected from polyamide, polyacrylonitrile, regenerated cellulose, and polyester is used as the fibrous material with an average diameter of 1 μm or more and 10 μm or less. Filter for leukocyte separation as described.