Name: stromal vascular fraction-enriched fat grafting for the treatment of symptomatic end-neuromata
Uploaded: 2017-11-23T13:00:00.000+00:00
Duration: 7 min 58 s
Description: Watch this Scientific Journal Video about Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata at JoVE.com

著者の謝辞があります。

プロトコル大学病院チューリッヒ人間研究倫理委員会ガイドラインに従います。
1. Neuromata と患者の選択の診断
<ol><li>術前神経ブロック、神経の近位を診断を確認するために実行します。前腕の中間の三番目のブロックを評価し、外側前腕皮神経 (LACN)、後骨間神経 (ピン)、尺骨神経の背側枝など、SBRN の神経支配領域の潜在的なトリガーを除外します。10。</li>
	<li>かなりの苦痛低減 (少なくとも 5 ポイントの痛みのための視覚アナログ スケール (VAS))11症例にのみ手術を行います。</li>
</ol>2. 手術の準備
<ol><li>どんな食べ物や液体摂取 6 h、手術前に患者を禁止します。</li>
	<li>ビーチ椅座位における普遍的な手術テーブルの中央手術室に患者を配置します。</li>
	<li>腕の残りに影響を受けた腕/手を配置します。</li>
	<li>気管内麻酔を行う麻酔科医を求める (フェンタニル 2 μ g/kg、ロクロニウム 0.6 mg/kg、プロポ フォール 150 250 mg)。</li>
	<li>脂肪吸引 (上下腹部や太ももの内側など) の領域と神経腫切除のサイトでポビドン ヨードで皮膚を消毒します。</li>
	<li>脂肪吸引の部位と手術部位を公開残して、滅菌外科用ドレープと患者をカバーします。</li>
</ol>3. 腫れ上がった脂肪吸引
<ol><li>脂肪吸引の領域で #11 刃メスで約 3-5 mm の直径のいくつかの小さな刺し傷 (穿刺) 切開を行います。皮下層に達する限り、切開します。</li>
	<li>皮下脂肪層 (深さ 3 mm) に希薄リドカインと (カニューレ浸潤) 14 ゲージ侵入者によるエピネフリン (500 mL 乳酸リンゲル液、20 mL の 1% リドカイン ・ エピネフリン 1 mg) を含む腫れ上がった溶液を注入して、脂肪採取の領域。</li>
	<li>腫れ上がったソリューションを有効にすることができます少なくとも 20 分待ちます。</li>
	<li>フック、3 mm 収集カニューレ (外径 3 mm、長さ 26 cm) 60 mL トーミー シリンジ、刺し切開で脂肪を収穫を開始。 注: 利き手は配置とカニューレのコースを継続的に制御します。</li>
	<li>60 mL マーク トーミー注射器のピストンを引いて、シリンジ内の負圧を維持し、支配的な手によって連続真空作成を避けるために撤退のプランジャーに沿って蚊クランプを配置します。</li>
	<li>トーミー注射器を提供トーミー シリンジ スタンドで縦置きに配置することで 220 mL の脂肪の最小値を収穫します。これは脂肪および流体からの分離の沈降をことができます。</li>
	<li>血の実質的な量なしで良質の脂肪移植を取得する特別な注意を取る。 メモ: は、コンテンツ拡張の血を気づいたときに脂肪吸引部を変更します。</li>
	<li>簡単なステッチで刺し切開を縫合し、圧力を適用必要に応じて (例えば圧縮帯)。</li>
	<li>免震システムの単回使用の滅菌使い捨てセットに、lipoaspirate を転送します。</li>
	<li>後 lipofilling の脂肪の割合を得るために付属のスタンドに lipoaspirate と 1 つのトーミー シリンジをしてください。</li>
</ol>4. 脂肪ティッシュの処理
<ol><li>滅菌の消耗品セットを閉じます。 メモ: 分離システム (材料の表を参照してください) ウェット テストを実行することにより完全性を確認します。</li>
	<li>画面の指示に従います。 注: プロセスは大抵ソフトウェア制御し、だけ「次へ」またはコントロール パネルの「戻る」ボタンを押して下記手順のマニュアルの確認が必要です。Lipoaspirate は組織コレクション キャニスターに移る。組織の収集容器に、lipoaspirate は体重し、残留血液細胞とぬれソリューションを削除するために乳酸リンゲル液で自動的に洗浄します。</li>
	<li>必要な場合より多くの脂肪を追加します。 注: 収穫の脂肪組織の重量に応じて分離システム要求より多くの脂肪を追加するユーザーまたは追加する追加酵素試薬の量を計算します。</li>
	<li>分離システムの指示に従って酵素試薬の必要量を追加します。 注: 酵素消化の過程で脂質と SVF の分離が実現されます。これにより、組織が振動運動です。酵素消化が完了した後、遠心室内の細胞濃度に SVF は自動的に転送します。SVF は細胞ペレットに集中して遠心室内 (遠心速度は約 3,000 x g)。最後に、細胞ペレットは、出力の残留酵素レベルを減らすように乳酸リンゲル液と洗浄のシリーズを受けます。プロセスが完了すると、透明な液体 (SVF) 5 mL が得られます。</li>
</ol>5. 手術
<ol><li>アプローチ、神経腫を皮膚切開を前腕の遠位 3 分の 1 で以前診断された神経腫サイトに譲る。</li>
	<li>手術用双眼ルーペ拡大鏡メガネ (2.5 から 3.5 X) の使用の下で皮下組織を解剖し、関与する神経、神経の近位に進みます。<ol>
<li>その後、神経を遠位に従うし、神経腫 (目視検査に基づく) を識別します。</li>
	</ol>
</li>
	<li>神経腫の完全な拡張が明確、まっすぐトランス ニューラルはさみカットによって、神経腫を切除する開始します。神経腫を識別しない場合は、神経の方法のみが実行されます。</li>
	<li>3 cm 近位手首まで残りの神経断端の方法に進みます。 注: 神経断端末はプリンシパルの切開の真ん中に配置されなければなりません。</li>
	<li>プリンシパルのアプローチ周りの表皮及び真皮の皮膚をメスで 2 ~ 4 小さな穿刺 (刺す) 切開 (直径 1 mm) を配置できます。</li>
	<li>2 ~ 4 鈍カニューレを配置 (16 ゲージ、長さ 9 cm) 切り株に刺し切開神経を介して。 注: カニューレの先端は神経断端に向かって監督する必要があります。</li>
	<li>粘着ドレッシングによって場所にカニューレを固定します。</li>
	<li>カニューレの周りステッチを配置し、ノードを確保することがなくその場で縫合糸を残してください。</li>
	<li>移植の流出を防ぐためにしっかりと主な切開部を縫合します。 注: は、カニューレがその場に残っていることを確認します。</li>
</ol>6. SVF 濃縮脂肪移植の応用
注: 処理の約 1.5 h 後、加工の SVF の 5 mL はそれ以上の使用の準備ができて。
<ol><li>SVF 通信 10 mL 注射器で 5 mL を吸引します。</li>
	<li>同じように反対の 10 mL の注射器で沈殿脂質画分の 2 mL を吸引します。</li>
	<li>1 つの注射器のピストンを引いてと通信、反対のシリンジのプランジャーを引っ張る十分な混合が得られるまで交互に堆積脂質画分の 2 mL と SVF の 5 mL を混ぜます。</li>
	<li>2 ~ 4 10 mL 注射器で均等に SVF 濃縮脂肪移植を分割します。</li>
	<li>神経の周りに均等に他は鈍カニューレに注射器を接続して切り株後、準備された SVF 濃縮脂肪移植の 1 つを配布します。 メモ: 移植を適用する前に、カニューレが正しい位置に固定されて、脂肪の生着を適用中、カニューレを脱臼しないように注意してください。</li>
	<li>カニューレを慎重に削除します。</li>
	<li>その後、steri-ストリップと小さな穿刺切開をテープします。</li>
	<li>組み込みのスプリント プラス (60-70 ° に屈曲中手指節 (MCP) 関節、趾節間 (IP) 関節を完全に拡張、拳投影で親指、25 ° 背屈で手首) の位置を適用することによって、手を固定します。神経の方法を実行すると、場合にのみ、添え木は必要なく、腕や手首を 10 日間の包帯します。</li>
</ol>7. 術後患者の取り扱い
<ol><li>後麻酔回復ユニット術後に患者を移すし、病棟の十分な監視を可能にします。</li>
	<li>手術後 10 日間、手首を固定し続けます。</li>
	<li>48 時間後手術後包帯の最初の変更を実行、定期的 (すなわち放電まで 1 日おき) で続行します。</li>
	<li>12-14 日術後縫合糸を削除します。</li>
	<li>2、6、12、フォロー アップ中の痛みと 36 ヶ月の臨床評価を行います。</li>
</ol>

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著者が明らかに何もありません。

本研究の目的は、組織的 SVF 濃縮脂肪移植症候性の終わり neuromata に対する新しい治療としての技術的な側面を説明するためにです。
SBRN の neuromata は、唯一の 3312の手術成功率に関連付けられます。Vaientiらは、早期フォロー アップの良好な結果のシリーズを発表神経脂肪移植 (PFG) 上肢5の痛み neuromata 8 ?...

症候性の終わり-neuromata、SBRN の管理のまま既存の外科的治療法の多くに挑戦です。主に、神経腫を切除し、筋肉1、2骨3、または静脈4など近隣の建物に移転すると共に残りの神経断端。これはアウト再生神経線維束終末器官の雰囲気を模倣して萌芽を防ぐためにされます。しかし、これらの技術のどれも証明に優れ、痛み再発と不十分な痛みの軽減率の高さが認められた多くの場合。
最近では、残りの神経断端周囲脂肪移植有望な結果が得られたし、上肢5の苦痛の終わり neuromata のための新しい処置の選択として提唱されました。継続的運動を防ぎ、痛みをタップする機械的な障壁として機能する以外脂肪移植の有益な側面はむしろ細胞レベルに起因します。ひと脂肪組織脂肪前駆細胞、マスト細胞、血管、内皮細胞、血管平滑筋細胞、線維芽細胞のなど、複数のセルが含まれているし、造血幹細胞6と一緒に脂肪由来幹細胞 (ADSCs)。この異種細胞人口脂肪由来再生細胞 (ADRCs) または「svf」、と呼ばれますと分離システムによって lipoaspirate から取得できます (材料の表を参照してください)、応用酵素試薬 (材料表を参照)7。アクションの正確なメカニズムはまだ完全に理解される;ただし、「svf」は、血管新生、抗炎症や分化、細胞外マトリックス8の分泌の改善に関連付けられています。
最近の研究で明らかに追加された SVF なし脂肪移植による 12 ヶ月後の痛み再発と SVF の濃縮細胞集団が吸収率を減らす脂肪移植の再生の潜在性を高める目的で脂肪の生着に追加されます。5します。 さらに、生物学的作用がまた切り株9と神経が絞扼される可能性がありますその後瘢痕 adherences のまとまりのない軸索発芽を減らすことによって神経自体に有益な効果を提供します。
SBRN の症候性の終わり neuromata の治療のための新規手術のオプションがされている SVF 濃縮脂肪移植を記述する最初の臨床研究は、最近10を公開しました。一般的には、テクニックは、偶発的または医原性神経損傷後に起こる、神経腫に適用できます。現在の後の研究の目的は、念入りにこの手順の技術的な側面を説明するためにです。

<table><tbody><tr><td>Cytori Celution System</td><td>Cytori</td><td></td><td>isolation system</td></tr><tr><td>Celase</td><td>Cytori</td><td></td><td>enyzme reagent</td></tr><tr><td>liposuction system</td><td></td><td></td><td></td></tr><tr><td>tumescent solution (500 ml Lactated Ringers, 20 ml of 1% lidocaine and 1 mg epinephrine)</td><td></td><td></td><td></td></tr><tr><td>14-gauge infiltrator</td><td></td><td></td><td></td></tr><tr><td>3 mm-collecting cannula (15, 26, 32 cm)</td><td></td><td></td><td></td></tr><tr><td>60 mL Tommey syringes</td><td></td><td></td><td></td></tr><tr><td>mosquito clamp</td><td></td><td></td><td></td></tr><tr><td>Toomey syringe stand</td><td></td><td></td><td></td></tr><tr><td>4 cannulas (fat injecting cannulas 1.5 mm, syringe NO.2, 90 mm)</td><td>Chalybs</td><td></td><td></td></tr><tr><td>10 mL syringes</td><td></td><td></td><td></td></tr><tr><td>connecting device for syringes</td><td></td><td></td><td></td></tr><tr><td>#11- and #15-blade scalpel</td><td></td><td></td><td></td></tr><tr><td>stevens scissors</td><td></td><td></td><td></td></tr><tr><td>pins pincette</td><td></td><td></td><td></td></tr><tr><td>bipolar electrocautery</td><td></td><td></td><td></td></tr><tr><td>suture material</td><td></td><td></td><td></td></tr><tr><td>adhesive dressing</td><td></td><td></td><td></td></tr><tr><td>binocular loupes magnifier glasses (2.5-3.5x)</td><td>Carl Zeiss</td><td></td><td></td></tr><tr><td>Statistical Package for Social Sciences, version 22</td><td>IBM</td><td></td><td></td></tr></tbody></table>

stromal vascular fraction-enriched fat grafting for the treatment of symptomatic end-neuromata

本研究の目的は、念入りに説明し、間質血管の端数 (SVF) の重要なステップを強調表示-の特に症候性の終わり neuromata 末梢感覚神経と本研究での新規治療薬として脂肪移植を豊かに橈骨神経 (SBRN) の表面的な枝。既存の処置の多数、にもかかわらず、永続的な術後疼痛と共通の痛み再発、まだ非常に一般的な評価手順の独立しました。神経腫、切除 microsurgically 適切に標準化されたプロトコルです。筋肉や骨などの解剖学的構造を近隣の再生神経断端の移転ではなく脂肪移植応用 perifocally、機械的な障壁として機能します。脂肪吸収率を減らし、グラフトの再生の可能性を高めるために、高濃度の SVF は接木で統合されています。SVF は、特定の商業免 lipoaspirate の酵素・ メカニックの分離法による皮下脂肪から分離されます。SVF 濃縮脂肪移植では、機械的な障壁と血管新生、炎症および線維化を改善など、細胞レベルで様々 な生物学的効果の両方を提供しています。機械的及び生物学的効果は、神経再生時に神経断端のまとまりのない軸索伸長を削減し、したがって苦痛の終わり neuromata の再発を防止するのに役立ちます。

データは、統計ソフトウェアを使用し、± 標準偏差を意味として分析されました。コルモゴロフ-スミルノフ検定研究人口のノンパラ メトリック分布を確認する適用し、負の経験分布を確認しました。研究人口のノンパラ メトリックの性質に基づき、関連のサンプル、ウィルコクソン検定は術前・術後の転帰を比較に採用されました。P 値の統計的有意性を考えた 0.05...

Watch this Scientific Journal Video about Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata at JoVE.com

Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata

間質血管分数濃縮脂肪移植症候性の終わり neuromata の治療のため

研究の目的は間質血管の端数 (SVF) の技術的なプロシージャを説明するために-症候性の終わり neuromata の新規治療薬として脂肪移植を濃縮します。この手法は、機械的な障壁と処理され、濃縮された SVF による細胞レベルの生物学的作用の両方の利点を提供します。

The purpose of this study was to methodically illustrate and highlight the crucial steps of stromal vascular fraction (SVF)-enriched fat grafting as a ...

stromal-vascular-fraction-enriched-fat-grafting-for-treatment

Research

JoVE Journal

Medicine

7.6K ビュー.  University Hospital Zurich. 研究の目的は間質血管の端数 (SVF) の技術的なプロシージャを説明するために-症候性の終わり neuromata の新規治療薬として脂肪移植を濃縮します。この手法は、機械的な障壁と処理され、濃縮された SVF による細胞レベルの生物学的作用の両方の利点を提供します。

ビデオ: Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata

A Novel Mammary Fat Pad Transplantation Technique to Visualize the Vessel Generation of Vascular Endothelial Stem Cells

Endothelial cells (ECs) are the fundamental building blocks of the vascular architecture and mediate vascular growth and remodeling to ensure proper vessel development and homeostasis. However, studies on endothelial lineage hierarchy remain elusive due to the lack of tools to gain access as well as to directly evaluate their behavior in vivo. To address this shortcoming, a new tissue model to study angiogenesis using the mammary fat pad has been developed. The mammary gland develops mostly in the postnatal stages, including puberty and pregnancy, during which robust epithelium proliferation is accompanied by extensive vascular remodeling. Mammary fat pads provide space, matrix, and rich angiogenic stimuli from the growing mammary epithelium. Furthermore, mammary fat pads are located outside the peritoneal cavity, making them an easily accessible grafting site for assessing the angiogenic potential of exogenous cells. This work also describes an efficient tracing approach using fluorescent reporter mice to specifically label the targeted population of vascular endothelial stem cells (VESCs) in vivo. This lineage tracing method, coupled with subsequent tissue whole-mount microscopy, enable the direct visualization of targeted cells and their descendants, through which the proliferation capability can be quantified and the differentiation commitment can be fate-mapped. Using these methods, a population of bipotent protein C receptor (Procr) expressing VESCs has recently been identified in multiple vascular systems. Procr+ VESCs, giving rise to both new ECs and pericytes, actively contribute to angiogenesis during development, homeostasis, and injury repair. Overall, this manuscript describes a new mammary fat pad transplantation and in vivo lineage tracing techniques that can be used to evaluate the stem cell properties of VESCs.

This work demonstrates a novel approach to assess the proliferation, differentiation, and vessel-forming potential of vascular endothelial stem cells (VESCs) through mammary fat pad transplantation followed by whole-mount tissue preparation for microscopic observation. A lineage tracing strategy to investigate the behavior of VESCs in vivo is also presented.

血管内皮幹細胞の血管形成を視覚化するための新規乳腺脂肪移植技術

Endothelial cells (ECs) are the fundamental building blocks of the vascular architecture and mediate vascular growth and remodeling to ensure proper ...

発生生物学

A Modified Heterotopic Swine Hind Limb Transplant Model for Translational Vascularized Composite Allotransplantation (VCA) Research

Vascularized Composite Allotransplantation (VCA) such as hand and face transplants represent a viable treatment option for complex musculoskeletal trauma and devastating tissue loss. Despite favorable and highly encouraging early and intermediate functional outcomes, rejection of the highly immunogenic skin component of a VCA and potential adverse effects of chronic multi-drug immunosuppression continue to hamper widespread clinical application of VCA. Therefore, research in this novel field needs to focus on translational studies related to unique immunologic features of VCA and to develop novel immunomodulatory strategies for immunomodulation and tolerance induction following VCA without the need for long term immunosuppression.
This article describes a reliable and reproducible translational large animal model of VCA that is comprised of an osteomyocutaneous flap in a MHC-defined swine heterotopic hind limb allotransplantation. Briefly, a well-vascularized skin paddle is identified in the anteromedial thigh region using near infrared laser angiography. The underlying muscles, knee joint, distal femur, and proximal tibia are harvested on a femoral vascular pedicle. This allograft can be considered both a VCA and a vascularized bone marrow transplant with its unique immune privileged features. The graft is transplanted to a subcutaneous abdominal pocket in the recipient animal with a skin component exteriorized to the dorsolateral region for immune monitoring.
Three surgical teams work simultaneously in a well-coordinated manner to reduce anesthesia and ischemia times, thereby improving efficiency of this model and reducing potential confounders in experimental protocols. This model serves as the groundwork for future therapeutic strategies aimed at reducing and potentially eliminating the need for chronic multi-drug immunosuppression in VCA.

A Modified Heterotopic Swine Hind Limb Transplant Model for Translational Vascularized Composite Allotransplantation VCA Research

Vascularized Composite Allotransplantations (VCA) have become a clinical reality. However, broad clinical application of VCA is limited by chronic multi-drug immunosuppression. The authors present a reliable and reproducible large animal model to translate novel immunomodulatory strategies that can minimize or potentially eliminate the need of immunosuppression in VCA.

トランスレーショナル血管柄コンポジット同種移植（VCA）研究用に修正異豚後肢移植モデル

Vascularized Composite Allotransplantation (VCA) such as hand and face transplants represent a viable treatment option for complex musculoskeletal trauma ...

医学

Assessment of Viability of Human Fat Injection into Nude Mice with Micro-Computed Tomography

Lipotransfer is a vital tool in the surgeon&#8217;s armamentarium for the treatment of soft tissue deficits of throughout the body. Fat is the ideal soft tissue filler as it is readily available, easily obtained, inexpensive, and inherently biocompatible.1 However, despite its burgeoning popularity, fat grafting is hampered by unpredictable results and variable graft survival, with published retention rates ranging anywhere from 10-80%. 1-3
To facilitate investigations on fat grafting, we have therefore developed an animal model that allows for real-time analysis of injected fat volume retention. Briefly, a small cut is made in the scalp of a CD-1 nude mouse and 200-400 &#181;l&#160;of processed lipoaspirate is placed over the skull. The scalp is chosen as the recipient site because of its absence of native subcutaneous fat, and because of the excellent background contrast provided by the calvarium, which aids in the analysis process. Micro-computed tomography (micro-CT) is used to scan the graft at baseline and every two weeks thereafter. The CT images are reconstructed, and an imaging software is used to quantify graft volumes.
Traditionally, techniques to assess fat graft volume have necessitated euthanizing the study animal to provide just a single assessment of graft weight and volume by physical measurement ex vivo. Biochemical and histological comparisons have likewise required the study animal to be euthanized. This described imaging technique offers the advantage of visualizing and objectively quantifying volume at multiple time points after initial grafting without having to sacrifice the study animal. The technique is limited by the size of the graft able to be injected as larger grafts risk skin and fat necrosis. This method has utility for all studies evaluating fat graft viability and volume retention. It is particularly well-suited to providing a visual representation of fat grafts and following changes in volume over time.

Fat grafting is an essential technique for reconstructing soft tissue deficits. However, it remains an unpredictable procedure characterized by variable graft survival. Our goal was to devise a mouse model that utilizes a novel imaging method to compare volume retention between differing techniques of fat graft preparation and delivery.

マイクロコンピュータ断層撮影とヌードマウスへのヒト脂肪注入の生存率の評価

Lipotransfer is a vital tool in the surgeon&#8217;s armamentarium for the treatment of soft tissue deficits of throughout the body. Fat is the ideal soft ...

A Patient-Derived Xenograft Model for Venous Malformation

Venous malformation (VM) is a vascular anomaly that arises from impaired development of the venous network resulting in dilated and often dysfunctional veins. The purpose of this article is to carefully describe the establishment of a murine xenograft model that mimics human VM and is able to reflect patient heterogeneity. Hyper-activating non-inherited (somatic) TEK (TIE2) and PIK3CA mutations in endothelial cells (EC) have been identified as the main drivers of pathological vessel enlargement in VM. The following protocol describes the isolation, purification and expansion of patient-derived EC expressing mutant TIE2 and/or PIK3CA. These EC are injected subcutaneously into the back of immunodeficient athymic mice to generate ectatic vascular channels. Lesions generated with TIE2 or PIK3CA-mutant EC are visibly vascularized within 7&#8210;9 days of injection and recapitulate histopathological features of VM patient tissue. This VM xenograft model provides a reliable platform to investigate the cellular and molecular mechanisms driving VM formation and expansion. In addition, this model will be instrumental for translational studies testing the efficacy of novel drug candidates in preventing the abnormal vessel enlargement seen in human VM.

We present a detailed protocol to generate a murine xenograft model of venous malformation. This model is based on the subcutaneous injection of patient-derived endothelial cells containing hyper-activating TIE2 and/or PIK3CA gene mutations. Xenograft lesions closely recapitulate the histopathological features of VM patient tissue.

静脈奇形に対する患者由来異種移植片モデル

Venous malformation (VM) is a vascular anomaly that arises from impaired development of the venous network resulting in dilated and often dysfunctional ...

Isolation of Murine Adipose Tissue-derived Microvascular Fragments as Vascularization Units for Tissue Engineering

A functional microvascular network is of pivotal importance for the survival and integration of engineered tissue constructs. For this purpose, several angiogenic and prevascularization strategies have been established. However, most cell-based approaches include time-consuming in vitro steps for the formation of a microvascular network. Hence, they are not suitable for intraoperative one-step procedures. Adipose tissue-derived microvascular fragments (ad-MVF) represent promising vascularization units. They can be easily isolated from fat tissue and exhibit a functional microvessel morphology. Moreover, they rapidly reassemble into new microvascular networks after in vivo implantation. In addition, ad-MVF have been shown to induce lymphangiogenesis. Finally, they are a rich source of mesenchymal stem cells, which may further contribute to their high vascularization potential. In previous studies we have demonstrated the remarkable vascularization capacity of ad-MVF in engineered bone and skin substitutes. In the present study, we report on a standardized protocol for the enzymatic isolation of ad-MVF from murine fat tissue.

We present a protocol to isolate adipose tissue-derived microvascular fragments that represent promising vascularization units. They can be rapidly isolated, do not require in vitro processing and, thus, may be used for one-step prevascularization in different fields of tissue engineering.

組織工学のための血管新生単位としてマウスの脂肪組織由来の微小血管フラグメントの単離

A functional microvascular network is of pivotal importance for the survival and integration of engineered tissue constructs. For this purpose, several ...

生物工学

Transplantation of Schwann Cells Inside PVDF-TrFE Conduits to Bridge Transected Rat Spinal Cord Stumps to Promote Axon Regeneration Across the Gap

Among various models for spinal cord injury in rats, the contusion model is the most often used because it is the most common type of human spinal cord injury. The complete transection model, although not as clinically relevant as the contusion model, is the most rigorous method to evaluate axon regeneration. In the contusion model, it is difficult to distinguish regenerated from sprouted or spared axons due to the presence of remaining tissue post injury. In the complete transection model, a bridging method is necessary to fill the gap and create continuity from the rostral to the caudal stumps in order to evaluate the effectiveness of the treatments. A reliable bridging surgery is essential to test outcome measures by reducing the variability due to the surgical method. The protocols described here are used to prepare Schwann cells (SCs) and conduits prior to transplantation, complete transection of the spinal cord at thoracic level 8 (T8), insert the conduit, and transplant SCs into the conduit. This approach also uses in situ gelling of an injectable basement membrane matrix with SC transplantation that allows improved axon growth across the rostral and caudal interfaces with the host tissue.

This article describes a technique to insert a hollow conduit between the spinal cord stumps after complete transection and fill with Schwann cells (SCs) and injectable basement membrane matrix in order to bridge and promote axon regeneration across the gap.

シュワンの移植細胞のギャップを渡って軸索の再生を促進するために切断のラット脊髄切り株をブリッジする PVDF ・ セッション導管内

Among various models for spinal cord injury in rats, the contusion model is the most often used because it is the most common type of human spinal cord ...

Mechanical Micronization of Lipoaspirates for Regenerative Therapy

Stromal vascular fraction (SVF) has become a regenerative tool for various diseases; however, legislation strictly regulates the clinical application of cell products using collagenase. Here, we present a protocol to generate an injectable mixture of SVF cells and native extracellular matrix from adipose tissue by a purely mechanical process. Lipoaspirates are put into a centrifuge and spun at 1,200 x g for 3 min. The middle layer is collected and separated into two layers (high-density fat at the bottom and low-density fat on the top). The upper layer is directly emulsified by intersyringe shifting, at a rate of 20 mL/s for 6x to 8x. The emulsified fat is centrifuged at 2,000 x g for 3 min, and the sticky substance under the oil layer is collected and defined as the extracellular matrix (ECM)/SVF-gel. The oil on the top layer is collected. Approximately 5 mL of oil is added to 15 mL of high-density fat and emulsified by intersyringe shifting, at a rate of 20 mL/s for 6x to 8x. The emulsified fat is centrifuged at 2,000 x g for 3 min, and the sticky substance is also ECM/SVF-gel. After the transplantation of the ECM/SVF-gel into nude mice, the graft is harvested and assessed by histologic examination. The result shows that this product has the potential to regenerate into normal adipose tissue. This procedure is a simple, effective mechanical dissociation procedure to condense the SVF cells embedded in their natural supportive ECM for regenerative purposes.

Here, we present a protocol to obtain stromal vascular fraction from adipose tissue through a series of mechanical processes, which include emulsification and multiple centrifugations.

成人再生療法のための機械的微細化

Stromal vascular fraction (SVF) has become a regenerative tool for various diseases; however, legislation strictly regulates the clinical application of ...

Preclinical Model of Hind Limb Ischemia in Diabetic Rabbits

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular disease is the development of ischemia. In severe cases, patients can develop critical limb ischemia in which they experience constant pain and an increased risk of limb amputation. Current therapies for peripheral ischemia include bypass surgery or percutaneous interventions such as angioplasty with stenting or atherectomy to restore blood flow. However, these treatments often fail to the continued progression of vascular disease or restenosis or are contraindicated due to the overall poor health of the patient. A promising potential approach to treat peripheral ischemia involves the induction of therapeutic neovascularization to allow the patient to develop collateral vasculature. This newly formed network alleviates peripheral ischemia by restoring perfusion to the affected area. The most frequently employed preclinical model for peripheral ischemia utilizes the creation of hind limb ischemia in healthy rabbits through femoral artery ligation. In the past, however, there has been a strong disconnect between the success of preclinical studies and the failure of clinical trials regarding treatments for peripheral ischemia. Healthy animals typically have robust vascular regeneration in response to surgically induced ischemia, in contrast to the reduced vascularity and regeneration in patients with chronic peripheral ischemia. Here, we describe an optimized animal model for peripheral ischemia in rabbits that includes hyperlipidemia and diabetes. This model has reduced collateral formation and blood pressure recovery in comparison to a model with a higher cholesterol diet. Thus, the model may provide better correlation with human patients with compromised angiogenesis from the common co-morbidities that accompany peripheral vascular disease.

We describe a surgical procedure used to induce peripheral ischemia in rabbits with hyperlipidemia and diabetes. This surgery acts as a preclinical model for conditions experienced in peripheral artery disease in patients. Angiography is also described as a means to measure the extent of introduced ischemia and recovery of perfusion.

糖尿病性ウサギにおける後肢虚血の前臨床モデル

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular ...

<meta charset="utf8"></head><body>再生医療のためのフィブリンハイドロゲル中の機械的間質血管分画

The Combination of Mechanically Isolated Stromal Vascular Fraction and Fibrin Hydrogel: A Processing Protocol

The regenerative potential of adipose-derived stromal cells (ASCs) has gained significant attention in regenerative and translational research. In the past, the extraction of these cells from adipose tissue required a multistep enzyme-based process, resulting in a heterogenous cell mix consisting of ACSs and other cells, which are jointly termed the stromal vascular fraction (SVF). More recently introduced mechanical SVF (mSVF) isolation protocols are less time-consuming and bypass regulatory concerns. We recently proposed a protocol that generates mSVF rich in stromal cells based on a combination of emulsification and centrifugation. One current issue in mSVF application for wound therapy application is the lack of a scaffold providing protection from mechanical manipulation and desiccation. Fibrin hydrogels have been shown to be a useful adjunct in cell transfer for wound healing purposes in the past. In the work herein, we delineate the preparation steps of an mSVF-fibrin hydrogel construct as a novel approach for translational research and clinical application.

<meta charset="utf8"></head><body>著者スポットライト:創傷治癒のための脂肪由来間質血管画分の可能性の探求

Mechanically isolated stromal vascular fraction (SVF) in combination with a fibrin hydrogel offers an easy and efficient carrier for viable adipose-derived stromal cells for various indications, including tissue engineering and or wound healing purposes. Here, we present the preparation of a mechanical SVF (mSVF)-fibrin hydrogel construct for translational research and clinical application.

機械的に単離されたStromal Vascular画分およびFibrinのヒドロゲルの組合せ:処理のプロトコール

The regenerative potential of adipose-derived stromal cells (ASCs) has gained significant attention in regenerative and translational research. In the ...

間質血管分数濃縮脂肪移植症候性の終わり neuromata の治療のため

要約

さらに動画を探す

この動画の章

血管内皮幹細胞の血管形成を視覚化するための新規乳腺脂肪移植技術

トランスレーショナル血管柄コンポジット同種移植（VCA）研究用に修正異豚後肢移植モデル

マイクロコンピュータ断層撮影とヌードマウスへのヒト脂肪注入の生存率の評価

静脈奇形に対する患者由来異種移植片モデル

組織工学のための血管新生単位としてマウスの脂肪組織由来の微小血管フラグメントの単離

シュワンの移植細胞のギャップを渡って軸索の再生を促進するために切断のラット脊髄切り株をブリッジする PVDF ・セッション導管内

成人再生療法のための機械的微細化

糖尿病性ウサギにおける後肢虚血の前臨床モデル

機械的に単離されたStromal Vascular画分およびFibrinのヒドロゲルの組合せ:処理のプロトコール

機械的に単離されたStromal Vascular画分およびFibrinのヒドロゲルの組合せ:処理のプロトコール

間質血管分数濃縮脂肪移植症候性の終わり neuromata の治療のため

要約

さらに動画を探す

この動画の章

血管内皮幹細胞の血管形成を視覚化するための新規乳腺脂肪移植技術

トランスレーショナル血管柄コンポジット同種移植（VCA）研究用に修正異豚後肢移植モデル

マイクロコンピュータ断層撮影とヌードマウスへのヒト脂肪注入の生存率の評価

静脈奇形に対する患者由来異種移植片モデル

組織工学のための血管新生単位としてマウスの脂肪組織由来の微小血管フラグメントの単離

シュワンの移植細胞のギャップを渡って軸索の再生を促進するために切断のラット脊髄切り株をブリッジする PVDF ・ セッション導管内

成人再生療法のための機械的微細化

糖尿病性ウサギにおける後肢虚血の前臨床モデル

機械的に単離されたStromal Vascular画分およびFibrinのヒドロゲルの組合せ:処理のプロトコール

機械的に単離されたStromal Vascular画分およびFibrinのヒドロゲルの組合せ:処理のプロトコール

シュワンの移植細胞のギャップを渡って軸索の再生を促進するために切断のラット脊髄切り株をブリッジする PVDF ・セッション導管内