JP5917392B2 - 組織及び細胞富化グラフトを最適化するシステム、方法及び組成物 - Google Patents
組織及び細胞富化グラフトを最適化するシステム、方法及び組成物 Download PDFInfo
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Description
本願は、2009年5月1日に出願された米国特許仮出願第61/174,860号(発明の名称”SYSTEMS, METHODS AND COMPOSITIONS FOR OPTIMIZING TISSUE AND CELL GRAFTS ”)の優先権主張出願であり、この出願を参照により引用し、その開示内容全体を本明細書の一部とする。
本明細書において提供される幾つかの実施形態は、自己由来移植手技、例えば自己由来移植において直接用いることができ又は移植に先立って細胞(例えば、脂肪由来再生細胞、脂肪由来幹細胞等)、添加剤等で強化できる乾燥又は脱水脂肪組織グラフトを作製する方法に関する。
本明細書において、補充され、増強され又は強化された脂肪組織グラフトを作製する方法及びシステムが開示され、例えば、この場合、グラフトは、未処理脂肪組織又は乾燥若しくは脱水脂肪組織である。例えば、本明細書において説明する幾つかの方法では、強化又は補充脂肪組織グラフトに追加の脂肪由来再生細胞又は脂肪由来幹細胞が補充され、例えば、リポダイジェステート又は再生細胞又は幹細胞から成る濃縮脂肪由来細胞集団が補充される。好ましくは、追加の脂肪由来再生細胞又は脂肪由来幹細胞は、同一の被験体から得られる。幾つかの実施形態では、追加の脂肪由来再生細胞又は脂肪由来幹細胞を別の被験体から得ることができる。
吸引脂肪組織は、10名のヒト被験体(N=10)から、リポサクション(N=5)、レーザ(N=3)又はボディジェット(Body-jet)採取(N=2)のいずれかの方法によって、医院で収集された。吸引した組織サンプルを、以下の4群へと無作為に割り当てた。(1)コントロール、(2)重力分離、(3)遠心分離、及び(4)PUREGRAFT (商標)組織グラフト調製物。コントロールサンプルはそれ以上の操作なしに直接解析した。重力分離群におけるサンプルを別にして60mL注射器中に10分間セットした。サンプルの液体部分を廃棄し、残存する脂肪組織を解析した。遠心分離群については、キャップした10mL注射器にサンプルを充填し、IEC固定角ローター遠心分離機の中へ置き、3000rpm(約1200g)で3分間遠心分離した。遠心後に脂肪組織の上部の遊離脂質を吸引によって除去し、下澄を排出させ、残存するグラフト組織を解析した。PUREGRAFT (商標)群におけるサンプルは、上述のシステム300(PureGraft (商標))中で洗浄した。概要を述べると、組織はシステム300の第1のチャンバの中へ上記のように導入される。組織を150mlの乳酸加リンゲル液により2回洗浄した。過剰な液体及び脂質をシステムの第2のチャンバから排出させるようにする。乾燥した組織を組織のエントリーポートを介して取り出し、結果解析に使用した。
豊胸手術を必要又は所望する患者を同定又は選択する。1単位の脂肪組織を患者から取り出し、好ましくは閉鎖された滅菌液体/組織経路を維持する脂肪由来幹細胞処理装置に提供した。例えば、カニューレは、滅菌液体/組織経路を維持しながら、脂肪由来幹細胞処理装置の組織収集容器又はチャンバ(例えば柔軟性袋)に連結される。確立された技法によってリポサクションを行って被験体から脂肪組織を取り出し、取り出した未処理の脂肪組織を組織収集容器/チャンバ中に引き込む。実質的に全ての生理的食塩水、赤血球、成熟脂肪細胞が除去されるまで、脂肪組織の第1の部分をPBSによってすすぎ洗いし(例えば、組織がもはや明らかに赤色でなくなるまで、連続して洗浄する)、洗浄排出物(廃液、例えば、生理的食塩水、赤血球、成熟脂肪細胞)は、閉鎖された滅菌液体/組織経路を維持しながら、組織収集容器又はチャンバに連結されるポートを介して無菌的に除去される。幾つかの実施形態において、廃液ポートは導管によって廃液収集容器に連結され且つ廃液収集容器は真空源及び導管に接合するように構成され且つ/或いは廃液収集容器は1つ又は複数の弁を含有することができる。次いで、洗浄した脂肪組織の第1の部分はさらなる処理のために組織収集容器/チャンバ中で保存するか、又は貯蔵容器又はチャンバに移すことができる。
実施例1において記載されるように、脂肪由来の再生力のある細胞を補充した脂肪の多いグラフトを調製する。脂肪由来再生力のある細胞を補充した脂肪グラフトを、被験体の顔面、臀部、胸部、又はふくらはぎに投与するか、又は任意の軟組織欠陥を補正するように投与する。
脂肪の多いグラフトを必要又は所望する患者を同定又は選択する。本明細書において記載されるか又は当該技術分野において公知であるように、1単位の脂肪組織を患者から取り出す。グラフトを最適化するために、システム300を配向し、システムに接続した廃液袋(図示せず)を床に降ろす。脂肪組織は、外部環境と第1のサブシステムの内部との間の連通を提供する組織接近ポート600を介して、システム300内に導入される。トーミー(Toomey)又は他の当技術分野で認められている注射器を使用して、ポートを介して脂肪吸引液を注入することができる。所望される量の組織が添加されるまで、第1のサブシステムの最大体積を越えないように注意して、これを反復することができる。いったん脂肪吸引液が添加されたならば、ドレーンチューブ上のピンチクランプ(図示せず)を閉鎖する。乳酸加リンゲル又は生理的食塩水等の洗浄溶液及び/又はすすぎ洗い溶液は、ポートを介するホイールクランプ(図示せず)を開くことによって又は滅菌IVチューブによってシステム300内に導入される。添加される洗浄溶液又はすすぎ洗い溶液の量は変動してもよい。一般的には、大多数の脂肪吸引液又は第1のサブシステムにおける他の組織が浸水するように、十分な洗浄溶液をシステム内に導入する。例えば乳酸加リンゲル液のような洗浄溶液を150ml添加する。次に、ホイールクランプを閉鎖し、システムは短時間(例えば15秒)の間、手作業で撹拌する。撹拌は、反転、回転、振動などの形態であり得る。システムは、振盪プラットホーム、オービタルシェーカなどの外部手段によって撹拌される。組織が完全に洗浄された後(観察又はあらかじめ確立した時間間隔によって決定されるように)、ドレーンピンチ弁(図示せず)又はポートを開き、重力下で、排出物は濾過材320及び分離スクリーン330を通過させる。或る特定の期間(例えば3分間)で排出物を排出させる。このサイクルは何回か繰り返すことができる。サイクルは合計4回洗浄のために反復してもよい。最後の洗浄サイクル後に、廃液を5〜10分間排出させる。一般的には、廃液の最大排出は約10分の排出後に起こる。したがって、機械設備の必要なしに、そして操作の簡単な閉鎖された滅菌システムにおいて、血液、腫脹液及び遊離脂質が実質的にない、より乾燥した洗浄された脂肪の多いグラフトが生成される。さらに、このプロセスはわずか20分で完了することができ、外科医は最終ステップ中のシステムの許容排水時間の変更によって水和の所望レベルを制御することができる。
当業者であれば、日常行われるに過ぎない実験を用いて、本明細書において説明した本発明の特定の実施形態の多くの均等例を認識し又は想到することができよう。かかる均等例は、以下の特許請求の範囲に記載された本発明に含まれるものである。
Claims (26)
- 脂肪組織処理装置であって、
第1の外側シェル及び第2の外側シェルを備えた柔軟性の押しつぶし可能な袋を有し、前記第1の外側シェル及び前記第2の外側シェルは、前記袋を形成さるように互いに接合され、
前記袋は、第1のサブシステムと連通状態にある第1のポートを備え、この第1のポートは、脂肪組織を受け入れ、前記袋の第1の外側シェルと前記袋内に配置されたフィルタとによって画定され、このフィルタは、複数の細孔を備え、これらの細孔は、液体、腫脹流体、赤血球、及び洗浄溶液を通過させ、且つ成熟した脂肪組織、再生細胞、幹細胞、前駆細胞及び結合組織を保持し、
前記袋は、更に、前記フィルタ及び前記袋の第2の外側シェルによって構成される第2のサブシステムと連通状態にある第2のポートを備え、前記第2のサブシステムは、更に、前記フィルタと前記袋の第2の外側シェルとの間に空間を形成するセパレータを備え、このセパレータは、前記フィルタの細孔よりも大きい細孔からなる分離スクリーン又はメッシュを備えていることを特徴とする脂肪組織処理装置。 - 前記セパレータは、前記第2のサブシステム内で自由浮動状態である、請求項1記載の装置。
- 前記第1の外側シェル及び前記第2の外側シェルは、二重ヒートシールによって互いに接合されている、請求項1記載の装置。
- 前記フィルタは、二重ヒートシールによって前記第1の外側シェル及び前記第2の外側シェルに接合されている、請求項3記載の装置。
- 前記セパレータは、メッシュからなる、請求項1記載の装置。
- 前記セパレータは、スクリーンからなる、請求項1記載の装置。
- 前記セパレータの孔径は、前記フィルタの前記孔径よりも大きく、或いはその5倍、6倍、7倍、8倍、9倍又は10倍以上である、請求項1記載の装置。
- 前記フィルタの前記孔径は、30μm以上である、請求項1記載の装置。
- 前記フィルタの前記孔径は、30μm〜200μmである、請求項1記載の装置。
- 前記セパレータは、300μm〜2000μmである大きさを備えた細孔を有する、請求項1記載の装置。
- 前記第1のポートは、アダプタに解除可能に連結されるよう構成され、前記アダプタは、注射器筒の先端部に解除可能に連結されるよう構成されている、請求項1記載の装置。
- 前記第1のポートは、物質が前記第1のポートに入ることができるが、前記第1のポートから出ることができないよう構成されている、請求項11記載の装置。
- 前記第1のポートは、変形可能なプラスチック弁を備えている、請求項12記載の装置。
- 前記注射器筒は、60mlカテーテル注射器筒である、請求項11記載の装置。
- 前記第1のポートは、滅菌流体/組織経路を維持した状態でカニューレに取り付けられるよう構成されている、請求項1記載の装置。
- 更に、前記第1のサブシステムと連通状態にある第3のポートを備え、この第3のポートは、前記袋の第1の外側シェルと前記袋内に配置されたフィルタとの間にある、請求項1記載の装置。
- 前記第3のポートは、Yコネクタに接続されている、請求項16記載の装置。
- 脂肪組織グラフトを作製する方法であって、
(a)前記未処理脂肪組織の第1の部分を請求項1記載の第1の装置の前記第1のサブシステム内に導入するステップと、
(b)生理学的洗浄溶液を前記第1のサブシステムに追加することによって前記未処理脂肪組織の第1の部分をすすぎ洗いするステップと、
(c)前記第1の装置の前記第2のサブシステムから、水、生理学的洗浄溶液、血液、遊離脂質又はこれらの任意の組み合わせから成る群から選択された流体を除去し、それにより前記脂肪組織を乾燥させるステップを有する、方法。 - 前記生理学的洗浄溶液は、乳酸加リンゲル液、ハートマン溶液、生理的食塩水及びPLASMALYTE(登録商標)溶液から成る群から選択される、請求項18記載の方法。
- ステップ(b)の前記未処理脂肪組織は、前記導入ステップに先立って、前記未処理脂肪組織の第1の部分の液体含有量の1/2、1/3又は1/4倍以下である液体含有量まで脱水する、請求項18記載の方法。
- ステップ(b)の前記未処理脂肪組織は、前記導入ステップに先立って、前記未処理脂肪組織の第1の部分の液体含有量の1/3倍以下である液体含有量まで脱水する、請求項18記載の方法。
- 一集団をなす脂肪由来再生細胞を脂肪組織の第2の部分から単離し、ステップ(c)の前記乾燥脂肪組織を前記脂肪由来再生細胞の前記単離された集団がステップ(c)の前記乾燥脂肪組織に浸透することができる条件下で前記脂肪由来再生細胞の前記単離された集団に接触させるステップを更に有する、請求項18記載の方法。
- 前記脂肪由来再生細胞の前記単離された集団は、ステップ(c)の前記乾燥脂肪組織を接触させるステップに先立って、遠心分離作用を受けない、請求項22記載の方法。
- 前記脂肪由来再生細胞の前記単離された集団は、請求項1記載の第2の装置の前記第1のチャンバ内に存在する脂肪組織を前記脂肪由来再生細胞を遊離させる条件下でコラゲナーゼに接触させることにより前記第2の装置内で調製される、請求項22記載の方法。
- ステップ(c)の前記乾燥脂肪組織を前記脂肪由来再生細胞の前記単離された集団と接触させるステップが前記第1の装置内で実施される、請求項24記載の方法。
- 脂肪由来再生細胞の単離された集団を調製するために使用されることを特徴とする請求項1乃至17の何れか1項に記載の装置。
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WO2010127310A1 (en) | 2010-11-04 |
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AU2010242780B2 (en) | 2016-04-21 |
KR101644534B1 (ko) | 2016-08-01 |
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CN102458302A (zh) | 2012-05-16 |
EP2424463B1 (en) | 2016-12-28 |
AU2010242780A1 (en) | 2011-12-01 |
BRPI1009981A2 (pt) | 2016-03-15 |
JP2012525157A (ja) | 2012-10-22 |
KR20160091454A (ko) | 2016-08-02 |
CA2760574C (en) | 2020-10-27 |
CA2760574A1 (en) | 2010-11-04 |
CN102458302B (zh) | 2015-11-25 |
KR20120020143A (ko) | 2012-03-07 |
EP2424463A4 (en) | 2014-02-26 |
HK1169301A1 (zh) | 2013-01-25 |
EP3150166A1 (en) | 2017-04-05 |
MX339769B (es) | 2016-06-08 |
KR101738324B1 (ko) | 2017-05-19 |
US9133431B2 (en) | 2015-09-15 |
AU2016204537A1 (en) | 2016-07-21 |
ES2625893T3 (es) | 2017-07-20 |
US9695398B2 (en) | 2017-07-04 |
EP2424463A1 (en) | 2012-03-07 |
AU2016204537B2 (en) | 2018-07-19 |
US20100279405A1 (en) | 2010-11-04 |
US20150353891A1 (en) | 2015-12-10 |
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