M.-A. Deutsch is supported by Dr. Rusche Forschungsprojekt (2014) Deutsche Stiftung f&#252;r Herzforschung and Deutsche Gesellschaft f&#252;r Thorax- Herz- und Gef&#228;&#223;chirurgie. M. Krane is supported by Deutsche Stiftung f&#252;r Herzforschung (F/37/11), Deutsches Zentrum f&#252;r Herz Kreislauf Forschung (DZHK B 13-050A; DZHK B 14-013SE) and Deutsche Forschungsgemeinschaft (KR3770/7-1; KR3770/9-1)

全ての実験は、ドイツのハートセンターミ​​ュンヘン、バイエルン、ドイツ（Gzを。55.2-1-54-2532.3-17-13）の政府によって承認されました。動物は、特定病原体フリーの施設に収容し、医学研究のための国民の協会と全米科学アカデミーが提供する「実験動物の管理と使用に関する指針」が策定した「実験動物管理の原則」に準拠した人の治療を受けました科学と国立衛生研究所（NIH出版番号86から23は、1985年改訂）で発表しました。 注：2つの湾曲鉗子と血管dilatators含む無菌の顕微機器の完全なセットは、手順（ 図2a）のために必要です。周術期の抗生物質による予防は、日常的に必要ではないが、免疫不全動物で示すことができます。動物をイソフルラン吸入端末を使用して屠殺しました。 ドナーの調製および移植片の収穫 &lt;オール&gt; <li>腹腔内（ip）ミダゾラムの注射（5.0ミリグラム/キログラム）、メデトミジン（0.5ミリグラム/ kg）およびフェンタニル（0.​​05 mg / kg体重）によって動物を麻酔。術野に仰臥位にマウスを置き、ペダル引っ込め反射により外科的な許容範囲を確認してください。 </li><li>クロルヘキシジンで皮膚を3回消毒し、腹部正中切開を行います。下大静脈を露出させるために左に濡れたガーゼで腸を撤回。 </li><li>抗凝固のために下大静脈を通って0.4ミリリットルの生理食塩水で希釈したヘパリンの50 IUを注入。ヘパリン化した後、容器1分を切開。 </li><li>ダイアフラムを開き、心臓を露出するための二国間の開胸術を行います。前胸壁を上方に反射し、クランプで所定位置に保持されます。 </li><li>胸腺を取り外し、露骨大動脈と肺動脈幹の間の結合組織を解剖。右上大静脈を切開。 </li><li>のレベルで動員上行大動脈を穿刺30 G針で腕頭動脈を5 mlシリンジに取り付けられ、ゆっくりと4℃の冷たい心停止液3mlで過度の圧力なしretrogradually心を灌流。冠循環への空気の侵入を避けます。さらに心停止を強化するために、移植片の上に氷冷生理食塩水をドロップします。 </li><li>腕頭動脈の原点に近位大動脈を横断します。 </li><li> 8-0シルクで心臓に近い劣ると優れたvenaの複数形の両方cavaeを連結し、合字に遠位にそれらを横断します。 </li><li>遠位に可能な限り肺動脈幹を自由に解剖。 8-0絹糸で分岐に近い肺動脈を結紮し、遠位にそれらを横断します。 </li><li>単一8-0絹結紮とすべて一緒に肺静脈を結紮し、それらを横断します。 </li><li>ドナーから心臓を取り出し、移植まで氷冷心停止液に保管してください。 </li></ol> 2.受信者の準備<ol><li>準備11番メスまたは顕微鏡下ではさみでポリイミドチューブを切断することにより吻合のための袖口。カフは、円筒状の本体と、血管クランプを配置するための拡張機能（円周の 1/3）、1ミリメートル（ 図2b）の各々の長さで構成されています。 </li><li>動物を麻酔し、上記のように外科的寛容性を確認します。水分補給のための0.25ミリリットル温かい生理食塩水を皮下（SC）を注入します。 </li><li>電気シェーバーを使用して、右頸部の毛を削除し、暖かいパッド上に仰臥位で動物を配置します。呼吸が損なわれる可能性がこのよう前肢を過延伸回避しながらテープのストランドと足を固定します。乾燥を防ぐために、眼軟膏を使用して、目を覆います。クロルヘキシジンで手術部位を3回消毒します。 </li><li>頸静脈ノッチに右下顎角から横皮膚切開を行います。 </li><li>ぶっきらぼうに内線を動員ernal頸静脈。バイポーラ鉗子で側枝を焼灼し、それらを横断します。 </li><li>顎下腺からの静脈との合流点のレベルで合字の間に外頸静脈を分割します。 </li><li>カフを介して外部頸静脈を引いて、カフの延長上に血管クランプを配置することによって、近位血管を閉塞。合字を削除し、1時10分、ヘパリン化生理食塩水で血管内腔を灌漑。 </li><li>カフの上に血管壁をエバート、プリセット周8-0絹糸結紮で固定します。 </li><li>胸鎖乳突筋の表面的な部分を削除します。 </li><li>右総頸動脈を動員し、頸動脈分岐下の合字間の血管を横断。 </li><li>カフを介して頸動脈を引いて、カフの延長上にYasargilクランプを配置することによって、近位血管を閉塞。合字を削除し、1時10分、ヘパリン化生理食塩水で血管内腔を灌漑。 </li><li>ゆっくりDILAT電子血管dilatatorと動脈内腔は、カフの上に血管壁を裏返すと、プリセット周8-0絹糸結紮で固定します。 </li><li>グラフト移植のための十分なスペースを得るために、顎下腺の右葉を削除します。 </li><li>移植まで湿った術野を保管してください。 </li></ol> 3.グラフト移植<ol><li>逆さまに内側に向い大動脈と横方向に肺動脈をレシピエント頸部におけるグラフトを配置します。 </li><li>頸動脈カフの上にグラフトの大動脈を引き、円周8-0シルク結紮で吻合を固定します。 </li><li>その前面に肺動脈幹を切開し、外頸静脈とカフの上に引き出します。再び周8-0絹糸結紮で吻合を固定します。 </li><li>頸動脈のアンクランプすることにより、以下、外頸静脈にクランプを取り外します。心臓は血液ですぐにいっぱいに。 Rを強化するために、eperfusionは、移植片に37℃の温かい生理食塩水をドロップします。通常、心臓は1分以内に洞調律を開発しています。静脈サイトでカフの拡張子を削除します。 </li><li>正しいグラフト位置を確認し、吻合は、ねじれのない緊張フリー、船でなければなりません。これは創傷治癒を損なう可能性があるため、皮膚の血管を焼灼しないでください。出血は通常、自然に停止します。シングル8-0連続縫合で皮膚を閉じます。 </li></ol> 4.術後ケア<ol><li>水分補給のための0.25ミリリットル温かい生理食塩水を皮下注射します。 </li><li>麻薬は、ナロキソン（1.2ミリグラム/キログラム）、フルマゼニル（0.​​5ミリグラム/ kg）を、それが胸骨横臥位を維持するのに十分な意識を取り戻したまで暖かいパッドの上に動物を左SC atipamezol（2.5 mg / kg体重）を注入拮抗します。任意の異常行動を登録するには、少なくとも1つの付加的な時間のための動物を観察します。完全にリ・まで、他の動物の会社に手術を受けた動物を返しません。赤。 </li><li>処置後72時間の最小持続時間のためのローカルの制度のプロトコルに従ってブプレノルフィン（1日に3回皮下0.05ミリグラム/ kg）または代替鎮痛で先制鎮痛を提供します。 </li><li>外科医と獣医師は動物を定期的に表示されるはずです。 &gt; 15％の体重減少の場合、無関心では、膨潤または外科側感染動物は、イソフルラン吸入端末を使用して安楽死させます。 </li></ol>

<ol>
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N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+graft+function+in+rodent+models+of+heart+transplantation.">Assessment of graft function in rodent models of heart transplantation.</a> Microsurgery. 28, 565-570 (2008).</li><li>Schramm, R., 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=The+subepicardial+microcirculation+in+heterotopically+transplanted+mouse+hearts:+an+intravital+multifluorescence+microscopy+study.">The subepicardial microcirculation in heterotopically transplanted mouse hearts: an intravital multifluorescence microscopy study.</a> J Thorac Cardiovasc Surg. 134, 210-217 (2007).</li><li>Liu, F., Kang, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterotopic+heart+transplantation+in+mice.">Heterotopic heart transplantation in mice.</a> J Vis Exp : JoVE. 238, (2007).</li><li>Gong, W., 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=Introduction+of+modified+cervical+cardiac+transplant+model+in+mice.">Introduction of modified cervical cardiac transplant model in mice.</a> Exp Clin Transplant. 10, 158-162 (2012).</li><li>Matsuura, A., Abe, T., Yasuura, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simplified+mouse+cervical+heart+transplantation+using+a+cuff+technique.">Simplified mouse cervical heart transplantation using a cuff technique.</a> Transplantation. 51, 896-898 (1991).</li><li>Chen, H., Zhang, Y., Zheng, D., Praseedom, R. K., Dong, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+kidney+transplantation+in+mice:+technique+using+cuff+for+renal+vein+anastomosis.">Orthotopic kidney transplantation in mice: technique using cuff for renal vein anastomosis.</a> PloS one. 8, e77278 (2013).</li><li>Maglione, M., 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=A+novel+technique+for+heterotopic+vascularized+pancreas+transplantation+in+mice+to+assess+ischemia+reperfusion+injury+and+graft+pancreatitis.">A novel technique for heterotopic vascularized pancreas transplantation in mice to assess ischemia reperfusion injury and graft pancreatitis.</a> Surgery. 141, 682-689 (2007).</li><li>Sucher, R., 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=Orthotopic+hind-limb+transplantation+in+rats.">Orthotopic hind-limb transplantation in rats.</a> J Vis Exp : JoVE. , (2010).</li><li>Kamada, N., Calne, R. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+the+rat.+Technique+using+cuff+for+portal+vein+anastomosis+and+biliary+drainage.">Orthotopic liver transplantation in the rat. 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The authors have nothing to disclose.

我々は、様々な実験的な設定で、90％以上の成功率で200以上の心臓移植を完了しました。ここに記載された技術では、完全な手順は、経験豊富な外科医によって60分以内に行うことができます。臓器獲得の前に子宮頸部の容器を調製することにより、総グラフト虚血時間は20分に制限することができます。移植のための時間（ウォーム虚血時間）縫合技術16に比べカフ技術を使用して大...

心臓移​​植モデルは、頻繁にI / R傷害、同種移植片拒絶反応、移植後血管障害および免疫調節剤の効果を調査するために使用されます。マウスモデルは、既知の免疫学的および遺伝的背景、多くの近交系およびトランスジェニック株および比較的低い実験コストの可用性のような多くの利点を兼ね備えています。 1973年には、マウスでの腹部心臓移植の技術は、最初のコリーら 1で説明しました。このモデルでは移植片は、レシピエントの腹部大動脈へと下大静脈への主肺動脈の上行大動脈の端側吻合によって血管再生されます。 1991年には、マウスでの子宮頸心臓移植のための縫合技術は、Chenら2によって説明されました。このモデルでは、逆行性冠動脈灌流は、GRの上行大動脈に受信者の頸動脈の吻合によって確立されました後方。静脈血の右心房、右心室に冠状静脈洞を介したドレインとは、レシピエント外頚静脈（ 図1）への肺動脈を通って排出されます。腹部グラフト移植と比較して、子宮頸モデルは、低侵襲性であり、移植片の表面的な場所は、追加のフォローアップ検査（ 例えば、触診、心エコー検査、生体蛍光顕微鏡）3,4のために非常に容易にアクセスが可能になります。 技術改良5,6にもかかわらず、このモデルのより広範な使用は、吻合漏れや血栓症の発生率が高いと小血管の縫合吻合を実行するための技術的な難しさに限定されていました。 1991年に、松浦ら 。血管壁が大幅に子宮頸心臓移植7のモデルを容易に合成シリンダー、上で裏返されたカフ技術を導入しました。技術はFUましたrtherは腎8、膵臓9、手足10、肝11,12、肺13,14および血管移植15を含む様々な実験移植モデルに採用。縫合技術と比較して、吻合のための所要時間と、従って、暖かい虚血時間が短く、カフ法16を用いて、より少ない血管合併症が大幅にあります。また、高い成功率を使用して操作を実行するためにマイクロサージェリーに少しのトレーニングで外科医を可能にします。カフ技術の欠点は、脳灌流を変更することができる総頸動脈の前提条件ライゲーションです。 このビデオの論文では、マウスの子宮頸心臓移植のための改良されたと単純化されたカフ手法を提示します。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Forceps JFC-7.2</td>
			<td>S&#38;T</td>
			<td>P-00036</td>
			<td>Curved tip 0.20 mm</td>
		</tr>
		<tr>
			<td>Vessel dilatator D-5a</td>
			<td>S&#38;T</td>
			<td>S-00124</td>
			<td></td>
		</tr>
		<tr>
			<td>Adventita scissor</td>
			<td>S&#38;T</td>
			<td>S-00102</td>
			<td></td>
		</tr>
		<tr>
			<td>Vascular clamp B-1V</td>
			<td>S&#38;T</td>
			<td>S-00396</td>
			<td></td>
		</tr>
		<tr>
			<td>Yasargil Clip</td>
			<td>Peter Lazic</td>
			<td>65,097</td>
			<td>3.5 mm lenght</td>
		</tr>
		<tr>
			<td>Bipolar forceps</td>
			<td>Micromed</td>
			<td>148-100-011</td>
			<td>tip 0.25 mm</td>
		</tr>
		<tr>
			<td>Polyimide tubes</td>
			<td>River Tech Medical</td>
			<td></td>
			<td>19-24 gauge</td>
		</tr>
		<tr>
			<td>8.0 silk ligatures</td>
			<td>Catgut</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Custodiol HTK solution</td>
			<td>Essential Pharmaceuticals</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

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.

私たちのチームは成功し、約20のトレーニング手術後に記載された技術を実行することができます。最も挑戦的な部分は、それが簡単にバックフリップと複数の試みが壁の引き裂きにつながる可能性があるため、カフの上に頸動脈の血管壁を裏返すことです。訓練期間の後、平均総手術時間は60±8分でした。 20分ごろ、ドナーの収穫のために、子宮頸部の血管と吻合し、皮膚閉鎖のための15分?...

Watch this Scientific Journal Video about Heterotopic Cervical Heart Transplantation in Mice at JoVE.com

Heterotopic Cervical Heart Transplantation in Mice

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-cervical-heart-transplantation-in-mice

Research

JoVE Journal

Medicine

11.1K ビュー.  German Heart Center Munich, Technische Universität München. 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.

ビデオ: Heterotopic Cervical Heart Transplantation in Mice

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

マウスで制御子宮裂傷の傷害

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

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 ...

Cardiac Catheterization in Mice to Measure the Pressure Volume Relationship: Investigating the Bowditch Effect

Animal models that mimic human cardiac disorders have been created to test potential therapeutic strategies. A key component to evaluating these strategies is to examine their effects on heart function. There are several techniques to measure in vivo cardiac mechanics (e.g., echocardiography, pressure/volume relations, etc.). Compared to echocardiography, real-time left ventricular (LV) pressure/volume analysis via catheterization is more precise and insightful in assessing LV function. Additionally, LV pressure/volume analysis provides the ability to instantaneously record changes during manipulations of contractility (e.g., &#946;-adrenergic stimulation) and pathological insults (e.g., ischemia/reperfusion injury). In addition to the maximum (+dP/dt) and minimum (-dP/dt) rate of pressure change in the LV, an accurate assessment of LV function via several load-independent indexes (e.g., end systolic pressure volume relationship and preload recruitable stroke work) can be attained. Heart rate has a significant effect on LV contractility such that an increase in the heart rate is the primary mechanism to increase cardiac output (i.e., Bowditch effect). Thus, when comparing hemodynamics between experimental groups, it is necessary to have similar heart rates. Furthermore, a hallmark of many cardiomyopathy models is a decrease in contractile reserve (i.e., decreased Bowditch effect). Consequently, vital information can be obtained by determining the effects of increasing heart rate on contractility. Our and others data has demonstrated that the neuronal nitric oxide synthase (NOS1) knockout mouse has decreased contractility. Here we describe the procedure of measuring LV pressure/volume with increasing heart rates using the NOS1 knockout mouse model.

This article describes the measurement of murine left ventricular function via pressure/volume analysis at different heart rates.

ボーディッチ影響を調べる：圧力体積関係を測定するために、マウスにおける心臓カテーテル検査

Animal models that mimic human cardiac disorders have been created to test potential therapeutic strategies. A key component to evaluating these ...

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 ...