This work was funded by the American Association of Plastic Surgeons 2014 Academic Scholar Award.

すべての手術の手順は、ジョンズ・ホプキンス大学と米国農務省および公衆衛生局の要件に準拠して完了しました。このプロトコルは、ジョンズ・ホプキンス大学動物実験委員会を以下の、施設内倫理委員会は、（プロトコル番号M013M490）のガイドラインを承認しました。最終的な生存データは、以下に記載の外科的処置のために記録しました。両方のドナーとレシピエント動物は0.1ミリグラムでブプレノルフィンを使用して先制麻酔を受ける/ kgの皮下1時間前に手術し、レシピエント動物のブプレノルフィンでは、移植後に同じ用量で再投与し、最初の48時間で、必要に応じて再投与されます手術後。 1.ドナー同種移​​植片回復注：受信者の麻酔時間を最小限にするために、以前の受信者の移植よりも移植40分のドナー部分を開始し、同時に終了時間または少し耳を容易にするために受信者の準備対Lierの終了時間。 <ol><li>手順については、標準的な無菌顕微楽器や滅菌手袋を使用してください。私たちの研究室では、顕微楽器のオートクレーブ滅菌を使用しています。 </li><li> 4％でイソフルラン誘導気化器を使用して、ドナーマウス（雄）を麻酔。非外傷性の機械的なバリカンを使用すると、頸部、胸部、および腹部から毛を取り除きます。仰臥位で動物を置き、ノーズ​​コーンを介して1〜2％のイソフルランを維持します。定期的につま先のピンチ引っ込め反射を評価することによって、作業中は適切な麻酔を確認してください。 </li><li>皮膚切開の前に、広く滅菌綿棒を使用して、イソプロピルアルコールが続く防腐剤ポビドンヨードを適用することにより、手術を準備します。 </li><li>子宮頸部と腹部の皮膚を横切るハサミで浅横皮膚切開を開始します。 midaxillary線に沿って、左右両方の切開を接続します。 </li><li>顕微鉗子を使用して6-0絹縫合糸とはさみで外頚静脈を識別連結および分割するために、両側頸部を分析。その後、電気メスを使用して左右対称に、内頸静脈と総頸動脈を露出させるために胸鎖乳突筋筋肉を分割します。左側と右側総頸動脈およびバルクの方法で内頸静脈の下で6-0絹縫合糸を渡します。 注：彼らは縛られ、後にステップ1.9で分割されます。 </li><li>はっきり頸部の残りの添付ファイルを解放するためにハサミを使用して、気管の前方に位置するストラップの筋肉と関連した緩い疎性結合組織を、分割します。 </li><li>バイポーラ電気メスと鋭い解剖を使用して、鎖骨下血管を露出させ、連結する大胸筋や鎖骨（6-0絹縫合糸）を分割して近位に分割します。 </li><li>次に、静かに、動物の陰茎を把握し、撤退。ペニスの背に沿って、陰茎の背静脈を視覚化し、消毒イソプロピルアルコールで地域。 30 G針を使用して、背静脈から静脈内にヘパリンの3万台を注入し、ペニスを元の位置に戻って反動することができます。周囲の組織へのヘパリン溶液の部分的な漏れが発生することがあります。 </li><li>総頸動脈と頸静脈の周りに配置され、以前にバルクタイを使用して、左右対称に、連結と構造を分割します。 </li><li>次に横intrabdominal切開を行うためにハサミを使用しています。肝臓下の下大静脈を露出させ、肝臓下の下大静脈に冷たいユーロコリンズ心停止溶液2mlを注入する腸を骨抜き。前次のステップに進むにハートビートの肝変色や停止を可視化することにより、適切な注入を確認してください。 注：ユーロ・コリンズ液を我々の研究室で用意され、特定の試薬および器具の表を参照してください。 </li><li>使用はさみは、二国間diaph経由で胸腔内の空洞にアクセス露出した腹部からragmatic切開。肋間筋と肋骨を介して切開頭を拡張します。同時に胸の壁に沿って内胸船舶の保全性を確保しながら、心臓、胸腺、および大血管を露出し胸壁を反映しています。 </li><li>冷たいユーロコリンズ心筋保護液4mlで肝上下大静脈を注入します。 </li><li>大動脈のルートを特定し、下行大動脈に遠位にトレースします。急激に下行大動脈（最大長を保持）をカットします。 </li><li>肺動脈幹を特定し、（最大長さを維持し）、その分岐点のすぐ近位分けます。そして、冷たいユーロコリンズ心停止溶液2mlを使用して、肺動脈幹の内腔に軟質プラスチックチップカテーテルを配置することによって、肺動脈幹と心を洗い流します。 </li><li> 6-0絹縫合糸を用いて、ライゲーションおよび下大静脈、肺静脈の合流、および両側上大静脈のアクセサリ枝を分割します。そして、昇格気道を入力させないように注意しながら主茎気管支や気管に沿って添付ファイルから心臓の頭を解剖。シャープとバイポーラ電気メスを使用すると、完全にドナーマウスからそれを解放胸壁、胸腺、および心臓を解剖します。 </li><li>最後に、内胸の血管（ 図1A）を混乱させないように注意しながら、胸骨と横肋骨に沿って、はさみを使って、小さいサイズにex vivoでの同種移植片の胸壁をトリミング。血行再建以下の出血を最小限に抑えるために、osteomusculocutaneous胸骨の境界に沿って、バイポーラ電気焼灼を使用しています。 </li><li>受信者は、インセットのために準備されていない場合、ユーロコリンズ液を冷（4 Oの摂氏）の10ml中の同種移植片を配置します。受信者がインセットの準備ができている場合は、まっすぐに受信者術野への同種移植片を移します。 </li></ol> 2.受信者の準備注：受信者の麻酔時間を最小限にするために、別々の手術駅で前にドナー同種移​​植片の収穫が完了するまでの約40分を受信者の準備を開始します。 <ol><li>手順については、標準的な無菌の顕微機器の別個のセットと滅菌手袋を使用してください。 </li><li> 4％でイソフルラン誘導気化器を使用して、（男性または女性）、レシピエントマウスを麻酔。非外傷性の機械的なバリカンを使用すると、右の子宮頸部と胸部の毛を削除します。 </li><li>仰臥位と角度右上肢が若干下方に頭と右上肢の間に110度の角度を形成する際にマウスを置きます。ノーズコーンを介して1〜2％イソフルランの麻酔を維持します。 </li><li>綿の先端のアプリケーターを用いてマウスの眼に石油眼軟膏を置きます。皮膚切開の前に、広くイソプロピルアルコールが続く防腐剤ポビドンヨードを使用して、手術部位を準備します。 </li><li>はさみを使用して、右下BORに沿って正中線から皮膚切開を作ります下顎のデルと右胸部にinfero-横方向に切開を拡張します。微小血管鉗子で鈍的切開を使用して、軟組織および外から円周方向に自由に船舶により外頸静脈を動員。電気メスを使用して、すべてのブランチを分割し、同種移植のための空き領域に鋭い解剖し、電気メスを使用して、顎下腺の右葉を削除します。 </li><li>カフの上に裏返すために外頸静脈のに十分な長さを確認し、6-0絹縫合糸を用いて、静脈外頚を結紮。プレカットポリイミドカフの内腔を通って静脈に挿入し、所定の位置に血管カフ複合体を固定するためのブルドッグ微小血管クランプを使用しています。そして、近位エバートカフの上に、外頸静脈を分割し、10-0ナイロン縫合糸で所定の位置に固定し、はさみを使って。 （ 図1B） </li><li>総頸動脈を露出するようにバイポーラ電気メスと右胸鎖乳突筋を分割します。円周方向に暴徒頸部内の最遠位点に動脈頭をilize。これは、軟組織と周囲の外膜を除去するために、鉗子で血管の鈍的切開を用いて達成されます。 </li><li> 6-0絹縫合糸を用いて、ライゲーションおよび総頸動脈を分割。プレカットポリイミドカフの内腔を通って動脈をパスし、可能な限り胸郭入口に近いブルドッグ微小血管クランプで所定の位置に固定します。優しくエバートカフの上に、顕微拡張器を使用して血管を拡張、遠位血管を分割し、10-0ナイロン縫合糸で所定の位置に固定します。 （ 図1B） 注：特定の顕微拡張器は、特定の試薬および器具の表に記載されています。 </li></ol> 3.同種移植片挿入図<ol><li>逆さまや斜めの位置に受信者の子宮頸部領域内の同種移植片を配置するための標準的な滅菌機器および滅菌手袋を維持します。 </li><li>次に、AOR降順ドナーを配置受信者の動脈カフ構造体の上にチックルーメンと10-0ナイロン縫合糸（ 図1Cおよび1D）と所定の位置に固定します。 </li><li>ドナー肺動脈とレシピエントマウス（ 図1Cおよび1D）の裏返し外頸静脈カフ構造体との間の段差3.2とファッション同じ吻合を。 </li><li>まず静脈微小血管クランプ（外頸静脈クランプ）を削除し、動脈クランプ（総頸動脈クランプ）をリリース。動脈再灌流の間に、任意の出血に対処するための同種移植片の全体を検査します。出血が可視化される場合、血液の損失を最小限に抑え、バイポーラ電気メスを使用して、出血の原因を軽減するために、動脈クランプを再適用します。 </li><li>グラフトを点検し、止血を確認してください。リリースと完全に動脈微小血管クランプを取り外します。心腔の急激な体積膨張と瞬時に明らかになるであろう、再灌流の徴候を示し、aとbを待つために心を守って0.5から1分以内に開始するために食べます。心を潤すために温かい生理食塩水（35度摂氏）を使用します。 </li><li>吻合上のねじれや張力を誘発しないように、解剖学的位置に胸壁ドレープ。 6-0連続ナイロン縫合糸（ 図1E）を使用している手術創の皮膚を閉じます。 </li></ol> 4.術後ケア<ol><li>水分補給のためにすぐに手術後に0.3ミリリットルの生理食塩水を腹腔内の流体ボーラスを管理します。 </li><li>そして皮下にそれぞれ、痛みや感染症予防のためにブプレノルフィン（0.1mg / kg）およびエンロフロキサシン（5mgの/キログラム）を注入します。 </li><li>麻酔から覚めるまで加熱ランプの下で動物を置き、胸骨横臥に戻ります。リカバリ時には、適切な同種移植灌流を確保する同種移植のフィブリル化ハートビートを視覚化するために、首を点検。 </li><li>目を覚ましや臥位で一度、他のマウスの会社なし（別のケージにマウスを返します）それは、食物および水を自由に受け取ることができる場所。 、右上肢の一時的なマイナー限定的運動に、ケージの床の上にゼラチン食料源を残します。 </li><li>術後1時間レシピエントマウスを観察し、それが食物と水を自由に摂取を受け取ることができますし、活動や栄養摂取のための最初の24時間1日3回検査してケージ施設にそれを返します。最初の72時間の必要に応じて、一日2回皮下痛みと苦痛とブプレノルフィン（0.1mg / kg）で再用量の兆候がマウスを監視します。その後毎日動物を調べ、毎週、それらの重量を量ります。 </li><li>任意のマウスは、痛み、苦痛の徴候を表示したり、飼料摂取量を減少させた場合には獣医スタッフに相談してください。早期の安楽死を考えてみましょう（私たちのプロトコルで安楽死法は、頸椎脱臼に続いて、7分間のCO 2過剰摂取を採用）。 </li><li>同種移植心拍の停止はsにマウスを促す特定のエンドポイントとして定義されていますacrificed。 </li></ol>

The authors do not have any conflicts of interest or financial disclosures to declare.

含まれる同種移植の免疫学的調査に要因が、急性および慢性拒絶反応、直接的および間接的な抗原提示、受信者の感作、または混合キメラの誘導のメカニズムに限定されるものではない現象の多くがあります。19動物モデルはなってきました移植免疫学の研究のためのゴールドスタンダード、およびマウスモデルは、一般に、その低い相対獣医や住宅需要の減少、コスト、トランスジ...

Cardiac transplantation is the treatment of choice for end-stage heart failure. Both technical advancements and pharmacological innovations have propelled the field to early graft acceptance rates above 90%.12,13 Despite this, 60-80% 5-year graft survival is at a standstill and chronic rejection, characterized by transplant vasculopathy, remains inevitable.14-16 Furthermore, patients are subjected to multiple surgical procedures and lifelong immunosuppression, which are associated with chest wall deformities and medical sequelae and toxicities, respectively. The need for innovative approaches to extend allograft survival, minimize the immunosuppressive requirements, and offer reconstructive options for anatomical deformities is pressing.
Vascularized composite allotransplantation offers a unique strategy for improving heart transplant outcomes both from an immunological aspect as well as a reconstructive perspective.17 Vascularized composite allografts are also unique in a way that they have an inherent source of donor-derived hematopoietic stem cells which has shown a favorable ability to reduce immunosuppression and induce and sustain mixed chimerism.1-8 Additionally, co-transplantation of the thymus has shown to prolong survival of both, solid organ transplants and vascularized composite allografts.2,9-11 Combining these strategies with heart transplantation offers a novel solution to the aforementioned challenges facing heart transplantation.18
Murine models serve as excellent platforms for mechanistic in vivo investigation because of the availability of antibodies and well-defined inbred and knockout strains.19-21 Although heart transplantation in mice is commonly studied using a heterotopic intraabdominal microsurgical suture transplant model22-25, a heterotopic, cervical, non-suture cuff technique model has shown to be extremely replicable, reliable, and carries fewer rates of thrombosis.19,26,27 The goal of this study is to develop a heterotopic en bloc osteomyocutaneous chest wall, thymus, and heart transplant technique in mice to study the immunological mechanisms of combined solid organ and vascularized composite allotransplantation using a cervical non-suture cuff technique. This cluster allograft is perfused through the anastomosis of the donor descending aorta to the right common carotid artery and the donor pulmonary artery to the right external jugular vein. Preservation of the internal thoracic vessels and associated thymus branches is paramount to perfusing the chest wall (sternum, ribs, muscles, and skin) and thymus.

<table border="0">
	<tbody>
		<tr>
			<td>Euro-Collins Solution</td>
			<td></td>
			<td></td>
			<td>The solution is not commercially purchased but rather prepared in the laboratory. To make a 500ml solution add the ingredient listed below to a 330ml of double distilled water. Mix well, and then fill in the rest of the 170ml of double distilled water into the solution to a final volume of 500ml. 
			Ingredients: 1.02g KH2PO4, 3.66g K2HPO4, 0.56g KCl, 0.42g NaHCO3, and 17.52g of glucose.</td>
		</tr>
		<tr>
			<td>Suture</td>
			<td>Ethilon</td>
			<td>MWI 72667</td>
			<td>6-0 Ethilon</td>
		</tr>
		<tr>
			<td>Suture</td>
			<td>Microsurgery Instruments, Inc.</td>
			<td>10-0 Nylon</td>
			<td>10-0 Nylon</td>
		</tr>
		<tr>
			<td>Polyimide Cuff Vein (21G)</td>
			<td>Vention Medical</td>
			<td>141-0043</td>
			<td>http://www.ventionmedical.com/products-and-services/polyimide-tubing/</td>
		</tr>
		<tr>
			<td>Polyimide Cuff Artery (24G)</td>
			<td>Vention Medical</td>
			<td>141-0027</td>
			<td>http://www.ventionmedical.com/products-and-services/polyimide-tubing/</td>
		</tr>
		<tr>
			<td>Soft plastic tip catheter</td>
			<td>Terumo</td>
			<td>SR*OX2419CA</td>
			<td>24G x 3/4&#34;&#160;</td>
		</tr>
		<tr>
			<td>Microsurgical dilator</td>
			<td>S&#38;T</td>
			<td>D-5a.1</td>
			<td>Dilator, 11cm, FH, 0.1mm AT10d</td>
		</tr>
	</tbody>
</table>

a novel microsurgical model for heterotopic, en bloc chest wall, thymus, and heart transplantation in mice

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression must be pursued. Employing vascularized bone marrow transplantation and co-transplantation of the thymus have shown promise in this regard in various animal models.1-11 Vascularized bone marrow transplantation allows for the uninterrupted transfer of donor bone marrow cells within the preserved donor microenvironment, and the incorporation of thymus tissue with vascularized bone marrow transplantation has shown to increase T-cell chimerism ultimately playing a supportive role in the induction of immune regulation. The combination of solid organ and vascularized composite allotransplantation can uniquely combine these strategies in the form of a novel transplant model. Murine models serve as an excellent paradigm to explore the mechanisms of acute and chronic rejection, chimerism, and tolerance induction, thus providing the foundation to propagate superior allograft survival strategies for larger animal models and future clinical application. Herein, we developed a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique. The experience in syngeneic and allogeneic transplant settings is described for future broader immunological investigations via an instructional manuscript and video supplement.

長期生存を達成した同系のC57BL / 6移植。同種移植片の設計（ 図1）は、動物の生存の観点および進行中の同種移植片の生存を評価する能力の成功であることが判明しました。これは、実行可能な、アクティブな継続的な同種移植毛の成長を残りの上を覆う皮膚を介して実証された、およびハートビートは、視覚化と指触診で評価することができました。生存データは、同系移植さ...

Watch this Scientific Journal Video about A Novel Microsurgical Model for Heterotopic, En Bloc Chest Wall, Thymus, and Heart Transplantation in Mice at JoVE.com

A Novel Microsurgical Model for Heterotopic, En Bloc Chest Wall, Thymus, and Heart Transplantation in Mice

To study combined solid organ and vascularized composite allotransplantation, we describe a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique.

マウスにおける異所のための新しい顕微モデル、エン圏胸壁、胸腺、および心臓移植

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression ...

a-novel-microsurgical-model-for-heterotopic-en-bloc-chest-wall-thymus

Research

JoVE Journal

Medicine

9.2K ビュー.  Johns Hopkins University School of Medicine. To study combined solid organ and vascularized composite allotransplantation, we describe a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique.

ビデオ: A Novel Microsurgical Model for Heterotopic, En Bloc Chest Wall, Thymus, and Heart Transplantation in Mice

A Modified Method for Heterotopic Mouse Heart Transplantion

Mice are often used as heart transplant donors and recipients in studies of transplant immunology due to the wide range of transgenic mice and reagents available. A difficulty is presented due to the small size of the animal and the considerable technical challenges of the microsurgery involved in heart transplantation. In particular, a high rate of technical failure early after transplantation may result from recipient death and post-operative complications such as hind limb paralysis or a non-beating heart. Here, the complete technique for heterotopic mouse heart transplantation is demonstrated, involving harvesting the donor heart and its subsequent implantation into a recipient mouse. The donor heart is harvested immediately following in situ perfusion with cold heparinized saline and transection of the ascending aorta and pulmonary artery. The recipient operation involves preparation of the abdominal aorta and inferior vena cava (IVC), followed by end-to-side anastomosis of the donor aorta with the recipient aorta using a single running 10-0 microsuture and a similar anastomosis of the donor pulmonary artery with the recipient IVC. Following the operation the animal is injected with 0.6 ml normal saline subcutaneously and allowed to recover on a 37&#160;&#176;C heating pad. The results from 227 mouse heart transplants are summarized with a success rate at 48 hr of 86.8%. Of the 13.2% failures within 48 hr, 5 (2.2%) experienced hind limb paralysis, 10 (4.4%) had a non-beating heart due to graft ischemic injury and/or thrombosis, while 15 (6.6%) died within 48 hr.

A technique is demonstrated for the microsurgical procedure for heterotopic transplantation of hearts in mice, including simplified methods for donor harvesting and recipient vessel anastomosis.

異マウスハートTransplantion用に変更する方法

Mice are often used as heart transplant donors and recipients in studies of transplant immunology due to the wide range of transgenic mice and reagents ...

医学

Murine Heterotopic Heart Transplant Technique

It is now over forty years since this technique was first reported by Corry, Wynn and Russell. Although it took some years for other labs to become proficient in and utilize this technique, it is now widely used by many laboratories around the world. A significant refinement to the original technique was developed and reported in 2001 by Niimi. Described here are the techniques that have evolved over more than a decade in the hands of three surgeons (Plenter, Grazia, Pietra) in our center. These techniques are now being passed on to a younger generation of surgeons and researchers.
Based largely on the Niimi experience, the procedures used have evolved in the fine details - details which we will endeavor to relate here in such a way that others may be able to use this very useful model. Like Niimi, we have found that a video aid to learning is a priceless resource for the beginner.

The manuscript describes the steps required to perform the heterotopic heart transplant in the mouse.

ネズミ異心臓移植技術

It is now over forty years since this technique was first reported by Corry, Wynn and Russell. Although it took some years for other labs to become ...

Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed results. However, the cellular and molecular mechanisms of the neotissue development, valve thickening, and stenosis development are not researched extensively. To answer the above questions, we developed a murine heterotopic heart valve transplantation model. A heart valve was harvested from a valve donor mouse and transplanted to a heart donor mouse. The heart with a new valve was transplanted heterotopically to a recipient mouse. The transplanted heart showed its own heartbeat, independent of the recipient&#8217;s heartbeat. The blood flow was quantified using a high frequency ultrasound system with a pulsed wave Doppler. The flow through the implanted pulmonary valve showed forward flow with minimal regurgitation and the peak flow was close to 100 mm/sec. This murine model of heart valve transplantation is highly versatile, so it can be modified and adapted to provide different hemodynamic environments and/or can be used with various transgenic mice to study neotissue development in a tissue engineered heart valve.

In order to understand the cellular and molecular mechanisms underlying neotissue formation and stenosis development in tissue engineered heart valves, a murine model of heterotopic heart valve transplantation was developed. A pulmonary heart valve was transplanted to recipient using the heterotopic heart transplantation technique.

異心臓移植のマウスモデルを用いた肺動脈弁の移植

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed ...

Rat Heterotopic Abdominal Heart/Single-lung Transplantation in a Volume-loaded Configuration

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft involves anastomosis of donor inferior vena cava (IVC) to recipient IVC, and donor ascending aorta (Ao) to recipient abdominal Ao. The right upper and middle lung lobes are preserved and function as conduits for blood flow from right heart to left heart.
There are several advantages to using this technique, and it lends itself to a broad range of applications. Because the graft is transplanted in a configuration that allows for dyamic volume-loading, cardiac function may be directly assessed in vivo. The use of pressure-volume conductance catheters permits characterization of load-dependent and load-independent hemodynamic parameters. The graft may be converted to a loaded configuration by applying a clamp to the recipient&#8217;s infra-hepatic IVC. We describe modified surgical techniques for both donor and recipient operations, and an ideal myocardial protection strategy. Depending on the experimental aim, this model may be adapted for use in both acute and chronic studies of graft function, immunologic status, and variable ventricular loading conditions. The conducting airways to the transplanted lung are preserved, and allow for acute lung re-ventilation. This facilitates analysis of the effects of the mixed venous and arterial blood providing coronary perfusion to the graft.
A limitation of this model is its technical complexity. There is a significant learning curve for new operators, who should ideally be mentored in the technique. A surgical training background is advantageous for those wishing to apply this model. Despite its complexity, we aim to present the model in a clear and easily applicable format. Because of the physiologic similarity of this model to orthotopic transplantation, and its broad range of study applications, the effort invested in learning the technique is likely to be worthwhile.

We describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The transplant configuration results in a partially loaded graft circulation, allowing direct functional assessment. This model may be employed for acute or chronic studies of function and immunologic status of the transplanted graft.

ラット異腹部ボリュームにロードされた構成でのハート/シングル肺移植

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft ...

Heterotopic Cervical Heart Transplantation in Mice

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies this model by avoiding challenging suture anastomoses of small vessels thereby reducing warm ischemia time. In comparison to abdominal graft implantation the cervical model is less invasive and the implanted graft is easily accessible for further follow-up examinations. Anastomoses are performed by pulling the ascending aorta of the graft over the cuff with the recipient&#8217;s common carotid artery and by pulling the main pulmonary artery over the cuff with the external jugular vein. Selection of appropriate cuff size and complete mobilization of the vessels are important for successful revascularization. Ischemia-reperfusion (I/R) injury can be minimized by perfusing the graft with a cardioplegic solution and by hypothermia. In this article, we provide technical details for a simplified and improved cuff technique, which should allow surgeons with basic microsurgical skills to perform the procedure with a high success rate.

The cuff technique shortens and facilitates the model of heterotopic cervical heart transplantation in mice by avoiding technically challenging suture anastomoses of small vessels. The technical details shown in this video paper should allow researches to establish this model in their laboratories.

マウスにおける異子宮頸心臓移植

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies ...

Heterotopic Renal Autotransplantation in a Porcine Model: A Step-by-Step Protocol

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality of life when compared to dialysis. Although the last few decades have seen major improvements in patient outcomes following kidney transplantation, the increasing shortage of available organs represents a severe problem worldwide. To expand the donor pool, marginal kidney grafts recovered from extended criteria donors (ECD) or donated after circulatory death (DCD) are now accepted for transplantation. To further improve the postoperative outcome of these marginal grafts, research must focus on new therapeutic approaches such as alternative preservation techniques, immunomodulation, gene transfer, and stem cell administration.
Experimental studies in animal models are the final step before newly developed techniques can be translated into clinical practice. Porcine kidney transplantation is an excellent model of human transplantation and allows investigation of novel approaches. The major advantage of the porcine model is its anatomical and physiological similarity to the human body, which facilitates the rapid translation of new findings to clinical trials. This article offers a surgical step-by-step protocol for an autotransplantation model and highlights key factors to ensure experimental success. Adequate pre- and postoperative housing, attentive anesthesia, and consistent surgical techniques result in favorable postoperative outcomes. Resection of the contralateral native kidney provides the opportunity to assess post-transplant graft function. The placement of venous and urinary catheters and the use of metabolic cages allow further detailed evaluation. For long-term follow-up studies and investigation of alternative graft preservation techniques, autotransplantation models are superior to allotransplantation models, as they avoid the confounding bias posed by rejection and immunosuppressive medication.

Porcine models of organ transplantation provide an important platform to study mechanisms of organ preservation. This article describes a heterotopic porcine renal autotransplantation model, which allows investigating new approaches to improve the outcome of transplantation using marginal kidney grafts.

ブタモデルにおける異自家腎移植：ステップバイステップのプロトコル

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality ...

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization and for minimizing organ damage resulting from prolonged extracorporal circulation. The goal of this paper was to demonstrate a fully functional and clinically relevant model of cardiopulmonary bypass in a mouse. We report on the device design, perfusion circuit optimization, and microsurgical techniques. This model is an acute model, which is not compatible with survival due to the need for multiple blood drawings. Because of the range of tools available for mice (e.g., markers, knockouts, etc.), this model will facilitate investigation into the molecular mechanisms of organ damage and the effect of cardiopulmonary bypass in relation to other comorbidities.

This paper describes how to perform cardiopulmonary bypass in mice. This novel model will facilitate the investigation of the molecular mechanisms involved in organ damage.

マウス モデルにおける心肺バイパス: 新しいアプローチ

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure. Multiple small-animal models of heart transplantation have been used to study the acute and long-term effects of novel therapies. However, few are tested and demonstrated success in clinical trials. It is of critical importance to evaluate new therapies in a clinically relevant large-animal model for efficient and reliable translation of basic studies&#39;&#160;findings. Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and previously used to investigate novel cardioprotective strategies. This procedure focuses on acute ischemia-reperfusion injury and is a reliable method to investigate novel interventions which have been tested and validated in smaller experimental models, such as the murine model. We demonstrate its usefulness in assessing cardiac performance during the early post-transplantation period and other potential possibilities enabled by the model.

Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and utilized to investigate novel cardioprotective strategies.

オルソトピック心臓移植の前臨床ブタモデル

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart ...

A Simplified Model for Heterotopic Heart Valve Transplantation in Rodents

There is an urgent clinical need for heart valve replacements that can grow in children. Heart valve transplantation is proposed as a new type of transplant with the potential to deliver durable heart valves capable of somatic growth with no requirement for anticoagulation. However, the immunobiology of heart valve transplants remains unexplored, highlighting the need for animal models to study this new type of transplant. Previous rat models for heterotopic aortic valve transplantation into the abdominal aorta have been described, though they are technically challenging and costly. For addressing this challenge, a renal subcapsular transplant model was developed in rodents as a practical and more straightforward method for studying heart valve transplant immunobiology. In this model, a single aortic valve leaflet is harvested and inserted into the renal subcapsular space. The kidney is easily accessible, and the transplanted tissue is securely contained in a subcapsular space that is well vascularized and can accommodate a variety of tissue sizes. Furthermore, because a single rat can provide three donor aortic leaflets and a single kidney can provide multiple sites for transplanted tissue, fewer rats are required for a given study. Here, the transplantation technique is described, providing a significant step forward in studying the transplant immunology of heart valve transplantation.

This protocol describes a simple and efficient method for the transplantation of aortic valve leaflets under the renal capsule to allow for the study of alloreactivity of heart valves.

げっ歯類におけるヘテロトピック心臓弁移植のための簡略モデル

There is an urgent clinical need for heart valve replacements that can grow in children. Heart valve transplantation is proposed as a new type of ...

A Porcine Heterotopic Heart Transplantation Protocol for Delivery of Therapeutics to a Cardiac Allograft

Cardiac transplantation is the gold standard treatment for end-stage heart failure. However, it remains limited by the number of available donor hearts and complications such as primary graft dysfunction and graft rejection. The recent clinical use of an ex vivo perfusion device in cardiac transplantation introduces a unique opportunity for treating cardiac allografts with therapeutic interventions to improve function and avoid deleterious recipient responses. Establishing a translational, large-animal model for therapeutic delivery to the entire allograft is essential for testing novel therapeutic approaches in cardiac transplantation. The porcine, heterotopic heart transplantation model in the intraabdominal position serves as an excellent model for assessing the effects of novel interventions and the immunopathology of graft rejection. This model additionally offers long-term survival for the pig, given that the graft is not required to maintain the recipient's circulation. The aim of this protocol is to provide a reproducible and robust approach for achieving ex vivo delivery of a therapeutic to the entire cardiac allograft prior to transplantation and provide technical details to perform a survival heterotopic transplant of the ex vivo perfused heart.

We present a protocol for utilizing a normothermic ex vivo sanguinous perfusion system for the delivery of therapeutics to an entire cardiac allograft in a porcine heterotopic heart transplant model.

心臓同種移植への治療薬の送達のためのブタヘテロトピック心臓移植プロトコル

Cardiac transplantation is the gold standard treatment for end-stage heart failure. However, it remains limited by the number of available donor hearts ...