この作品は、オーストラリアのMicrosearch財団、およびMyeeコドリントン医学研究財団、オーストラリアプロジェクト無償1029205国立保健医療研究評議会（NHMRC）によってサポートされていました。

実験の開始前に、計画された実験のための関連機関の動物管理倫理委員会の承認を得る。あなたの機関の要件に従って、マウスを維持する。以下のプロトコルは、シドニー大学とロイヤルプリンスアルフレッド病院の委員会によって承認されている。 手術を開始する前に、すべての器具は徹底的に洗浄し、80％エタノールに浸すことにより滅菌されるべきである。いくつかの機関は、しかし、これは長期的に細かい顕微楽器を損傷する可能性があり、オートクレーブ処理を規定している。可能な場合に使用できる滅菌の使い捨て器具。 1。麻酔<ol><li>密閉容器内のイソフルランでマウスを麻酔し、素早く麻酔ノーズコーンを接続、操作盤の上に仰臥位に配置します。何レフがないことを確認するために後肢足をつまんで麻酔のテスト妥当。 </li><li>外科用メスで皮膚を剃ると80％エタノールで滅菌する。また、70％イソプロピルアルコール中2％クロルヘキシジンは、皮膚消毒のために使用することができる。麻酔導入時にイソフルラン濃度は3％ですが、メンテナンスのための百分の1から1.5まで絞り。定期的な呼吸や心拍​​数を維持するために、この濃度の微調整を行います。加熱パッド上に動物の温度を維持する。 </li></ol> 2ドナー操作<ol><li>胸壁フラップを形成するために、剣状突起のレベルで横のカットに続いて腋窩に肋骨端から胸郭の両側を通して胸を切断することにより、心臓や血管を露出するために開胸術を行います。頭の横にこのフラップを持ち上げて、オペレーティングボードにピン。心臓や血管を露出させ、心膜をはがします。 </li><li> IVCを通して心臓に冷たい、ヘパリン化生理食塩水の近位片手で、その他の注入1ミリリットルとピンセットで下大静脈を持ち上げ、その未然防止にIVCの小さな動脈クランプを置くバック針穴を通して灌流液のT流れ。 </li><li>ガーゼや綿棒を使って、上行大動脈と肺動脈を露出させ、下方の心を撤回。大動脈と肺動脈の束にチャンネル（横洞）事後を通してmicroscissorsの一方のブレードをパスし、吻合のための十分な長さを確保するために限り遠位に可能な限り一緒に大動脈と肺動脈を切った。 </li><li>ネクタイやIVC、右上大静脈（SVC）に分割は、6-0絹糸を使用して、SVCと肺静脈を左に。肺静脈と一緒に左のSVCを囲み、それを結ぶために心臓にシングルスレッドの後方に配置して、別々に、IVCと右のSVCを接続します。移植まで4℃で冷滅菌生理食塩水中に保管した後、タイへの遠位血管を切断することにより、ドナー部位から心臓を収穫します。これは放血によるドナーの死をもたらす。 </li></ol> 3。受信者の操作<ol><li>麻酔をかける（セクション1）上記のように受信者。慎重にリトラクターを形成するために曲げられ剣状突起とペーパークリップを使用して後退させる恥骨から正中切開により開腹を行い、その後、刺激を避けるために腹部を剃る。暖かい滅菌生理食塩水に浸したガーゼで腸をラップし、腹部の右上に撤回。 </li><li>インフラ腎大動脈と下大静脈を露出させ、焼灼装置を用いて、腰椎の血管から、それらを分割することにより腸骨分岐に左腎動脈と静脈から大動脈と下大静脈のバンドルされたセグメントを解放する。焼灼が正しい温度にあると血管端を分割して密封するのに十分な時間に使用することに注意してください。 </li><li>大動脈遠位及び近位にIVCセグメントに小さな非外傷性血管クランプを適用します。最初の30のG針で穴を穿刺することによって大動脈の前壁の切開をすることができる。その後、ドナーの大動脈の大きさに合わせてmicroscissors垂直に切開を切った。任意のBLを削除するには、ヘパリン化生理食塩水で大動脈内腔をフラッシュしますOODの塊。 </li><li>冷生理食塩水に浸したガーゼで覆われた受信者のサイトへのドナー心臓を持参し、腹部の右側に配置します。ドナーの大動脈が受容大動脈と受信者のIVCの隣に位置して、ドナーの肺動脈に切開の隣に位置していることを確認します。 </li><li>動物が180°回転した段階れる遠位の角に達するまで、第1の左側に、近位の角や縫合から始まる10-0ナイロン縫合糸を実行している使用して、受信者の大動脈にドナーの大動脈端側を吻合。穏やかに大動脈の右側を露出し、遠位端から近位端まで大動脈壁の右側を通って縫合継続する腹部の左側にドナー心臓を動かす。大動脈吻合を閉じる前に、静かに任意の血栓や空気を除去するために、ヘパリン化生理食塩水でルーメンをフラッシュします。 </li><li>受信者IVCドナー肺動脈（PA）の端側を吻合。切開バージョンを作るtically動脈吻合に従い、サイトでのIVCの前壁にmicroscissorsと。 IVCの内腔内に左側壁の遠位端から出発10-0ナイロン縫合糸を実行して、レシピエントにドナーIVC PAを吻合。近位端部に到達した後、吻合を完了するために遠位端に前方右側壁に沿って上昇縫合糸を続ける。吻合を閉じる前に、静かに任意の血栓や空気を除去するための内腔をフラッシュします。 </li><li>血管クランプを解放する前に、場所吻合サイトの周りゲルフォームの断片を、止血が達成されるまで綿棒で穏やかな圧力を適用します。血管再生の際には、近位のクランプが続き、第一の遠位のクランプを解除する。 </li><li>血行再建後は、その回復を支援するために、外部から移植片に37℃で温かい生理食塩水を適用します。移植片は、通常はすぐに細動を開始し、数分以内に洞調律に自然に戻ります。温かい0.6ミリリットルを注入レシピエントの水和を維持するために生理食塩水を皮下。手術が完了する前に鎮痛のためのブプレノルフィンの皮下に注入する</li><li>両方の層のための1 5-0吸収性ランニング縫合糸で腹部の創傷を閉じます。内層を完了することによって起動し、皮膚に沿って継続する。 </li></ol> 4。回収および移植片の監視<ol><li>感染症の予防のためのアンピシリンを注入し、回復のために、37℃で加熱パッド上で受信者を置く。ほとんどの動物は急速に回復し、通常は飲んで、多くの場合、3時間以内に食べている。マウスは苦痛の徴候を示していた場合は、原因を特定するために密接に調べます。明らかな原因があれば、ブプレノルフィンで処理し、密接に監視します。症状が重いか、8時間以上を持続する場合は、獣医師に相談して安楽死を考えてみましょう。必要な場合は、症状が解決するまで、ブプレノルフィンの12時間ごとに注射を継続して与える。マウスは8時間後に苦痛の徴候を示している場合は、12を続けて与える彼らが解決するまで、ブプレノルフィンの毎時注射。症状は以上48時間継続し、必要に応じて安楽死させる場合は、獣医師に相談してください。 </li><li>直接腹部触診により移植心拍を監視し、+ + + +のような健康的な原因の高度な拒絶に弱拍のための+への移植片とのために、ビートの強さを記録する - 移植片の拒絶反応を完了することにより、非拍動など。実験期間中、週3回、その後、最初の10日間、毎日、マウスを監視します。 </li></ol>

<ol>
	<li>Aramaki, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interleukin-10+but+not+transforming+growth+factor-beta+is+essential+for+generation+and+suppressor+function+of+regulatory+cells+induced+by+intratracheal+delivery+of+alloantigen.">Interleukin-10 but not transforming growth factor-beta is essential for generation and suppressor function of regulatory cells induced by intratracheal delivery of alloantigen.</a> Transplantation. 79, 568-576 (2005).</li><li>Chen, R. H., Bushell, A., Fuggle, S. V., Wood, K. J., Morris, P. 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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+technique+of+heart+transplantation+in+rats.">Improved technique of heart transplantation in rats.</a> J. Thorac. Cardiovasc. Surg. 57, 225-229 (1969).</li><li>Corry, R. J., Winn, H. J., Russell, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Primarily+vascularized+allografts+of+heart+in+mice.">Primarily vascularized allografts of heart in mice.</a> Transplantation. 16, 343-350 (1973).</li><li>Larsen, C. 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著者らは、開示することは何もありません。

マウスの心臓移植がマスターにかなりの外科技術を必要と挑戦的な顕微法である。最も困難な局面は、血管の小さい直径である。また、受信者動作時間と出血を制限する必要がある。マウス心臓移植のための技術は、第一のコリーらによって記載された。 1973年に、その後モットラムら 12による。私たちの修正は、以下の点が含まれています。まず、すぐに灌流後の大動...

臓器移植の動物モデルは、臨床移植患者の治療を改善するための貴重な情報を提供することができる。マウスモデルは、遺伝子改変マウスや他の動物モデル1-3には使用できませんマウスに特異的な試薬の広い範囲に臓器移植拒絶または受諾の免疫機構を研究するために特に有用である。移植のマウスモデルでの課題は、満足のいく成果を達成するために、かなりの技術的なスキルを必要とし、ドナーとレシピエントのサイズが小さいです。 技術が最初に、その後、コリーら 5によるマウス心臓移植のために適合したラット4で心の異所移植のために記述されていた。この手法は、移植された心臓またはtの生存を危うくする可能性があり、どちらも、前にドナー操作に受信者とドナー心臓の無灌流の準備に関与彼は、受信者。本来移植拒絶反応および耐性6-8のメカニズムを調べるために記載した方法が広く用いられている。その他は、マウス9,10における心臓移植のために小野とリンジーの元のラットの心臓移植の手順を適応している。さらに最近では、技術はコリーら 11の方法に関連する問題のいくつかを克服したマウスの心臓移植のために出版された。直ちに開胸後のインサイチュ灌流冷ヘパリン化生理食塩水および受信者の動作時間を最小限に抑えるために従来の受信者動作にドナー動作を行う。プロトコルは、ここで説明挙げモットラムら 12に記載の方法に基づいて、我々の修飾を組み込んでいる。加えて、我々は代わりに6-0絹のネクタイの小さな非外傷性血管クランプを使用しています。血管クランプはより多くのスペースを取ることの欠点を持っていますが、彼らはあまりにもタイトな緩みすべきではないネクタイ、より制御が容易であるそしてクランプよりも除去が少なく簡単です。初期学習代替ステージが、我々の方法は、血管吻合のための単一の実行中の縫合糸を使用して縫合糸の均一性を確保するために、近位および遠位の角に滞在縫合糸を使用し、その吻合部分の開通することです。

<table border="0" cellpadding="0" cellspacing="0" width="831">
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			<td height="21" style="height:21px;width:293px;">Operating microscope&#160;</td>
			<td style="width:247px;">Leica, Heerbrugg, Switzerland</td>
			<td style="width:124px;">M651</td>
			<td style="width:167px;">10x - 25x magnification</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Anesthetic machine&#160;</td>
			<td>Vet Quip Pty Ltd, Sydney, Australia</td>
			<td style="width:124px;">Vett3</td>
			<td>Capable of delivering a mixture of isoflurane and oxygen in air&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:293px;">Operating board&#160;</td>
			<td style="width:247px;">Hardware store or office supplier</td>
			<td style="width:124px;"></td>
			<td>Dense cork or synthetic capable of taking pins</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:293px;">Heparinised saline (5 U/ml, 4oC)</td>
			<td>Pfizer, USA</td>
			<td style="width:124px;">FW25</td>
			<td>In 1ml syringe with 23 gauge needle on ice</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:293px;">Normal saline (37oC)</td>
			<td style="width:247px;">AstraZeneca Pty Ltd, Australia</td>
			<td style="width:124px;">4538</td>
			<td>In 1ml syringe with 23 gauge needle on warm pad</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:293px;">Sutures 10- nylon, 5-0 Vicryl</td>
			<td>Ethicon, Inc. NJ, USA</td>
			<td style="width:124px;">2870G/J421H</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Buprenorphine (0.05mg/kg in 0.1ml saline)</td>
			<td>Reckitt Benckiser, Sydney, Australia</td>
			<td style="width:124px;"></td>
			<td>In 1ml syringe with 23 gauge needle on ice</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ampcillin (5mg/kg in 0.1 ml saline)&#160;</td>
			<td>Aspen, Sydney, Australia</td>
			<td style="width:124px;"></td>
			<td>In 1ml syringe with 23 gauge needle on ice</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Gel Foam&#160;</td>
			<td>Pharmacia &#38; Upjohn Co. USA</td>
			<td style="width:124px;">801289304</td>
			<td>Cut into small pieces</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">High temperature cautery device&#160;</td>
			<td>Medtronic, USA</td>
			<td style="width:124px;">8444000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Microsurgery instruments:</td>
			<td colspan="2">Shanghai Medical Instruments Co. Ltd.,&#160;&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;&#160;&#160;&#160;&#160;&#160;&#160; micro-needleholders</td>
			<td style="width:247px;">Shanghai, China</td>
			<td style="width:124px;">WT2020</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;&#160;&#160;&#160;&#160;&#160;&#160; micro scissors</td>
			<td>&#160;&#160;&#160;&#160; &#34;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; &#34;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; &#34;</td>
			<td style="width:124px;">WT1020</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:293px;">&#160;&#160;&#160;&#160;&#160;&#160;&#160; micro-forceps (straight tip)</td>
			<td style="width:247px;">&#160;&#160;&#160;&#160; &#34;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; &#34;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; &#34;</td>
			<td style="width:124px;">WA3010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;&#160;&#160;&#160;&#160;&#160;&#160; micro-forceps (curved tip)</td>
			<td>&#160;&#160;&#160;&#160; &#34;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; &#34;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; &#34;</td>
			<td style="width:124px;">WA3020</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;&#160;&#160;&#160;&#160;&#160;&#160; micro-mosquito clamps (1 pair)</td>
			<td style="width:247px;">&#160;&#160;&#160;&#160; &#34;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; &#34;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; &#34;</td>
			<td style="width:124px;">W40350</td>
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a modified method for heterotopic mouse heart transplantion

マウスはしばしば、利用可能なトランスジェニックマウスおよび試薬広範囲の移植免疫学の研究において、心臓移植のドナーとレシピエントとして使用されている。難しさに起因動物のサイズが小さく、心臓移植に関与する顕微かなりの技術的な課題に提示される。特に、初期の移植後の技術的な故障率の高さは、受信者の死や、後肢麻痺または非心臓の鼓動のような術後の合併症に起因する可能性がある。ここでは、異マウス心臓移植のための完全な技術は、レシピエントマウスにドナー心臓とその後の注入を収穫含む、実証されている。ドナー心臓は、上行大動脈と肺動脈の寒さのヘパリン化生理食塩水や離断とその場での灌流直後に収穫される。受信者動作は、続いて腹部大動脈および下大静脈（IVC）の調製を含むシングルランニング10-0 microsuture者と受信者のIVCとドナーの肺動脈の類似の吻合を使用して、受信者の大動脈とドナーの大動脈の端側吻合。操作以下の動物0.6mlの生理食塩水を皮下注射し、37℃の加熱パッド上で回復させる。 227マウス心臓移植の結果は86.8％の48時間での成功率で要約されている。 15（6.6％）が48時間以内に死亡した48時間以内で13.2％の失敗、5（2.2％）、虚血性傷害および/または血栓症を移植片に起因する非鼓動心を持っていた後肢麻痺、10（4.4％）を経験した。

初期トレーニング期間の後、我々の群におけるマウス異心臓移植の227例を分析した。最初の24時間以内で成功率は90.3％であり、48時間後に、それは86.8％であった。 48時間以内で30（13.2％）の障害、5（2.2％）後肢麻痺を経験し、安楽死させなければならなかった、10（4.4％）15ながら、虚血傷害および/または血栓症を移植片に起因する非鼓動心を持っていた（6.6％）48時間以内に死亡した。い?...

Watch this Scientific Journal Video about A Modified Method for Heterotopic Mouse Heart Transplantion at JoVE.com

A Modified Method for Heterotopic Mouse Heart Transplantion

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 ...

a-modified-method-for-heterotopic-mouse-heart-transplantion

Research

JoVE Journal

Medicine

11.9K ビュー.  University of Sydney. 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.

ビデオ: A Modified Method for Heterotopic Mouse Heart Transplantion

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

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are available to perform mechanistic in vivo studies. While heart transplantation models using a suture technique were first successfully developed in rats, the translation into an equally widespread used murine equivalent was never achieved due the technical complexity of the microsurgical procedure. In contrast, non-suture cuff techniques, also developed initially in rats, were successfully adapted for use in mice1-3. This technique for revascularization involves two major steps I) everting the recipient vessel over a polyethylene cuff; II) pulling the donor vessel over the formerly everted recipient vessel and holding it in place with a circumferential tie. This ensures a continuity of the endothelial layer, short operating time and very high patency rates4.
Using this technique for vascular anastomosis we performed more than 1,000 cervical heart transplants with an overall success rate of 95%. For arterial inflow the common carotid artery and the proximal aortic arch were anastomosed resulting in a retrograde perfusion of the transplanted heart. For venous drainage the pulmonary artery of the graft was anastomosed with the external jugular vein of the recipient5.
Herein, we provide additional details of this technique to supplement the video.

The murine cervical heart transplantation model is well suited for immunological as well as ischemia reperfusion injury studies. We modified the procedure using a non-suture cuff technique and performed more than 1,000 successful transplants with this approach.
Herein, we provide additional details of this technique to supplement the video.

変更されたカフ法を用いたマウス子宮頸心臓移植モデル

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are ...

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 ...

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.

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 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 ...

A Modified Cuff Technique for Mouse Cervical Heterotopic Heart Transplantation Model

Cardiac allograft rejection limits the long-term survival of patients after heart transplantation. A mouse heart transplantation model is ideal for investigating the mechanism of cardiac allograft rejection in preclinical studies because of their high homology with human genes. This understanding would help develop unique approaches to improving patients&#39; long-term survival treated with cardiac allografts. In a mouse model, abdominal donor heart implantation is commonly performed with an end-to-end&#160;anastomosis to the recipient&#39;s aorta and inferior vena cava using stitches. In this model, the donor&#39;s heart is implanted by end-to-end anastomosis to the recipient&#39;s carotid artery and jugular vein by the modified-Cuff technique. The transplantation surgery is performed without stitching and thus may increase the survival of the recipient since there is no interference with the blood supply and venous reflux of the lower body. This mouse model would help investigate the mechanisms underlying the immunological and pathological (acute/chronic) rejection of cardiac allografts.

In the present protocol, a mouse heart transplantation model is used for investigating the mechanism of cardiac allograft rejection. In this heterotopic heart transplantation model, operation efficiency is improved, and the survival of cardiac grafts is ensured by a cervical end-to-end anastomosis of heart implantation using a modified Cuff technique.

マウス頸部異所性心臓移植モデルのための修正カフ法

Cardiac allograft rejection limits the long-term survival of patients after heart transplantation. A mouse heart transplantation model is ideal for ...