JP4511922B2 - 血小板を血液から単離するための方法および装置 - Google Patents
血小板を血液から単離するための方法および装置 Download PDFInfo
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- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/25—Chemistry: analytical and immunological testing including sample preparation
- Y10T436/25375—Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
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Description
血小板の抽出
全血からの血小板の抽出は研究されていた(非特許文献1)。輸血薬品では、その意図は各々の患者に必要な成分だけを輸血することであるため、血液センターの目標は血液成分をできる限り純粋に、すなわち最も少ない汚染細胞を有する形態で、製造することである。血小板は単離および精製が最も難しい。非特許文献1からのデータに基づくと、最適な遠心条件下(低速における長期間)でも、血小板のかなりの部分が沈殿した赤血球の中に捕獲されたままである。
V=沈殿速度であり、
ω=回転の角速度であり、
R=血液細胞からローター中心までの放射方向距離であり、
d=比重であり、
r=血液細胞の半径であり、
ηt=t℃の温度における媒体の粘度である]
にほぼ従い遠心により促進させうる。
弱回転遠心
全血が低速(1,000gまで)で短時間(2〜4分間)にわたり遠心される場合には、白血球が赤血球から速く沈殿しそして両者とも血小板よりはるかに速く沈殿する(上記のスベドベルグによる)。より高い速度では、同じ分布がより短かい時間で得られる。これが、明白に分離されておらず且つ(1)大多数の懸濁された血小板並びに少量の白血球および赤血球を含有する血漿、並びに(2)その下にある大多数の白血球および一部の血小板と混合された赤血球の薄層よりなる血液成分の層を生成する。全血からの血小板に富んだ血漿(PRP)の回収方法はこの原理に基づく。遅い遠心後に全血中の血小板のほとんどが血漿の中にあって血漿中の血小板の濃度が高まるため、用語「血小板に富んだ」はこの成分に関して使用される。パック入り赤血球およびPRP中への分離までだけ分別を行う遠心沈殿を「弱回転」と称する。「弱回転」はここでは、赤血球は沈殿するが血小板は懸濁液中に残る遠心条件を記述するために使用される。「強回転」はここでは、赤血球が沈殿し且つ血小板が赤血球層のすぐ上にある層の中に沈殿する遠心条件を記述するために使用される。
2回転血小板分離
弱回転後に、PRPを赤血球層から分離容器に除去することができ、そして第二遠心段階でPRPを血小板が少ない血漿(PPP)および血小板濃縮物(PC)に分別することができる。第二回転で、血小板は一般に遠心されて少量の血漿中より後で再懸濁させるペレットにされる。
強回転遠心
遠心が低速で続けられる場合には、白血球は赤血球の頂部に沈殿するが血小板は血漿中に懸濁されて残るであろう。長期にわたる低速遠心後にのみ血小板も赤血球の頂部に沈殿するであろう。
血小板収率および遠心速度
いわゆる「バフィコート」は血小板および白血球の層よりなるが、普通は血漿層の下方部分および赤血球物体の上層と共に回収される。この用途では、血小板層に関する全ての言及は白血球が存在しない場合には血小板層を意味しまたは白血球が血小板と混合されて存在する場合にはバフィコート層を意味することを意図する。
アフェレシス−単一回転血小板分離
アフェレシス、すなわち他の成分をドナーに再注入しながらの血液からの血小板の分離、を行うための特殊な装置が発明されていた。これにより、赤血球の損失がドナーが与えうる血液の量を制限するため、二段階遠心で可能なものより多い血小板をドナーが与えうる。典型的には、2〜3時間のアフェレシス工程は従来の供血の6倍以上に相当する3×1011個の血小板を含有する血小板生成物を生ずるであろう。
ロイコリダクション(Leukoreduction)
研究室2回転処理およびアフェレシス方法の両者から生ずるPCはドナー白血球を含有する。白血球は血小板貯蔵に悪影響を与えそしてサイトキン(cytokine)生成により輸血後に副作用を与えうることが示された。非自己白血球(異種白血球)およびそれらが製造するサイトキンが受容者の白血球により激しい反応を引き起こしうるため、PRPおよびPCからの白血球の除去(ロイコリダクション)が主要な問題である。1999年に、FDA血液生成物勧告委員会(FDA Blood Product Advisory Committee)は米国における全ての非白血球成分の慣例的ロイコリダクションを推奨した(非特許文献3)。従って、非自己由来白血球が有害な免疫反応を刺激するため、先行技術の多くは血小板濃縮物のロイコリダクションに焦点をあてている。本発明の方法は患者の血液から自己由来血小板を迅速に回収するための簡便な方法を提供するため、免疫反応は危険ではなく、そして白血球の存在はほとんどまたは全く関係ない。
自己由来血小板
自己由来血小板は異種血小板と比べて利点を有することが示された。異種または外来性調剤と関連する疾病伝染および免疫学的反応に関する事象は最少にされる。自己由来調剤が手術時に製造されることが、研究室システムにより起きうる試料の誤標識に関連する危険性を減ずる。自己由来血小板の使用は時間のかかるスクリーニング試験の必要性を回避する。血小板活性化を行う時間がより少ない。部分的に活性化される貯蔵された血小板とは異なり、自己由来血小板の活性化状態は、最初に製造された時点で、元の全血中のものと同様であることが見出された(非特許文献4)。
ピエテルツ(Pietersz)著、2000 デウィット(deWit)著、1975 ホルム(Holme)著、2000 クラウサー(Crawther)著、2000 ナイトン(Knighton)著、1986 マークス(Marx)著、1998 ゲーリング(Gehring)著、1999 マン(Man)著、2001 オズ(Oz)著、1992 ストーバー(Stover)著、2000
図1に描写されたタイプのパラソルデザイン血小板濃縮装置を組み立てた。フロートはポリエチレンおよびポリカーボネートから、1.06g/mlの全密度を有するような割合で、構成された。フロートの外径は2.62cmでありそしてその全長は4.57cmであった。フロートを、血小板受容器空洞内にある2個の直径0.32cmのステンレス鋼ボールと一緒に、軟質シリコーンゴム管の密封された端部の中に挿入した。軟質管は2.54cmの内径、0.08cmの壁厚さ、および密封された遠位端部を有していた。フロートを含有する軟質管は、2.86cmの内径および11.43cmの長さを有する硬質ポリカーボネート管の中に囲まれていた。軟質管の頂部は硬質管の頂部の上で折られそして7.62cm管23(図1参照)を有するキャップが溝12とかみ合う管23を有する軟質管の折られた頂部の上に適合された。
図9に描写されたタイプの血小板濃縮装置を組み立てた。フロートはポリエチレンおよびポリカーボネートから、1.08g/mlの全密度を有するような割合で、構成された。フロートの外径は2.535cmでありそしてその全長は1.2cmであった。フロートを、2.540cmの内径および11.43cmの長さを有する硬質ポリカーボネート管の中に挿入した。硬質管の底を密封した。
プランジャー下の空間とプランジャー上の空間との間の流体連結だけが血小板受容器空洞によるようにするためにスノーケル管を用いないこと以外は、図9に描写されたタイプの血小板濃縮装置を組み立てた。フロートはポリエチレンおよびポリカーボネートから、1.08g/mlの全密度を有するような割合で、構成された。フロートの外径は2.535cmでありそしてその全長は1.2cmであった。フロートを、2.540cmの内径および11.43cmの長さを有する硬質ポリカーボネート管の中に挿入した。硬質管の底を密封した。
Claims (4)
- 縦の内表面のある空洞を有する遠心回転分離容器と空洞内に配置されたフロートを含んでなる血液血小板分離装置であって、フロートが基部および基部の上にある血小板収集表面を有し、フロートが外表面を有し、フロートが赤血球の密度より低く且つ血漿の密度より高い密度を有し、そして血小板収集表面がフロート上に遠心後にそれを血小板の高さのすぐ下に置く高さに配置され、血小板収集表面が、凹面である円錐形である、血液血小板分離装置。
- 空洞が円筒状内表面を有し且つフロートが円筒状外表面を有する、請求項1の血液血小板分離装置。
- フロートの外表面と管の内表面との間の距離が0.5mmより小さい、請求項2の血小板分離装置。
- フロートの外表面が空洞の内表面と滑動関係にある、請求項1の血液血小板分離装置。
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US37755902P | 2002-05-03 | 2002-05-03 | |
US37995102P | 2002-05-10 | 2002-05-10 | |
US38263902P | 2002-05-21 | 2002-05-21 | |
PCT/US2003/012888 WO2003092894A2 (en) | 2002-05-03 | 2003-04-24 | Method and apparatus for isolating platelets from blood |
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JP2005524451A JP2005524451A (ja) | 2005-08-18 |
JP4511922B2 true JP4511922B2 (ja) | 2010-07-28 |
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US (8) | US20030205538A1 (ja) |
EP (1) | EP1509326B1 (ja) |
JP (1) | JP4511922B2 (ja) |
AT (1) | ATE364446T1 (ja) |
AU (1) | AU2003265160B2 (ja) |
CA (1) | CA2483931C (ja) |
DE (1) | DE60314413T2 (ja) |
ES (1) | ES2289306T3 (ja) |
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Cited By (1)
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JP2010527912A (ja) * | 2007-04-12 | 2010-08-19 | バイオメット・バイオロジックス・リミテッド・ライアビリティ・カンパニー | ブイ式懸濁液分画システム |
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Publication number | Priority date | Publication date | Assignee | Title |
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JP2010527912A (ja) * | 2007-04-12 | 2010-08-19 | バイオメット・バイオロジックス・リミテッド・ライアビリティ・カンパニー | ブイ式懸濁液分画システム |
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ES2289306T3 (es) | 2008-02-01 |
US20130294983A1 (en) | 2013-11-07 |
US7470371B2 (en) | 2008-12-30 |
WO2003092894A3 (en) | 2004-03-25 |
CA2483931C (en) | 2012-11-27 |
DE60314413D1 (de) | 2007-07-26 |
EP1509326A2 (en) | 2005-03-02 |
US20070034579A1 (en) | 2007-02-15 |
US20050186120A1 (en) | 2005-08-25 |
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EP1509326B1 (en) | 2007-06-13 |
DE60314413T2 (de) | 2008-02-21 |
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AU2003265160A1 (en) | 2003-11-17 |
US20110065183A1 (en) | 2011-03-17 |
CA2483931A1 (en) | 2003-11-13 |
US20090101599A1 (en) | 2009-04-23 |
US8950586B2 (en) | 2015-02-10 |
US20030205538A1 (en) | 2003-11-06 |
WO2003092894A2 (en) | 2003-11-13 |
AU2003265160B2 (en) | 2009-05-07 |
JP2005524451A (ja) | 2005-08-18 |
US8474630B2 (en) | 2013-07-02 |
US20050196874A1 (en) | 2005-09-08 |
US20120230890A1 (en) | 2012-09-13 |
US7223346B2 (en) | 2007-05-29 |
ATE364446T1 (de) | 2007-07-15 |
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