この作品は、CKBのNIH（RO1 HL098228）からの助成金によって部分的にサポートされていました。

注：すべての動物の手続きは、全国の小児病院施設内動物管理使用委員会によって承認された。 心臓弁ドナーマウスから1。肺心臓弁の収穫<ol><li>手術前に、すべての手術器具をオートクレーブ：細かいハサミ、3Xマイクロ鉗子、2倍のマイクロ血管クランプ、鉗子を適用する1Xクランプ、1Xマイクロニードルホルダー、1X春のはさみ、1Xリトラクターを1X。 </li><li> 6〜8週齢の雌C57BL / 6マウスは、肺、心臓弁供与体として使用される。そのケージからマウスを取り外して重量を量ると、ケタミン/キシラジンカクテル（ケタミン、200 mg / kgのキシラジンと、20 mg / kgを、IP）の過剰投与により安楽死させる。 </li><li>胸の部分をクリップし、パッドの上に背側横臥位にマウスを置きます。その後、開胸を行います。 、心臓を露出右心房に小さなカットを行い、氷冷生理食塩水で左心室を灌流。 </li><li>ぶっきらぼう昇順Aからの肺動脈（PA）を解剖するオルタ。肺動脈の2ミリメートルカフと一緒に肺動脈弁（PV）を切り出す。心の残りを処分。 </li><li>冷たいヘパリンおよび食塩水（100単位/ ml）で、PVを格納します。注：PVは、ドナー心臓への移植前2時間溶液中に保持することができる。 </li></ol> 2。ドナー心臓の準備<ol><li> 6〜8週齢の雌C57BL / 6マウスは、心臓ドナーとして使用される。そのケージからマウスを取り外して重量を量ると、ケタミン/キシラジンカクテル（ケタミン、200 mg / kgのキシラジンと、20 mg / kgを、IP）の過剰投与により安楽死させる。これは、端末の手順である。 </li><li>胸の部分をクリップし、パッドの上に背側横臥位にマウスを置きます。その後、開胸を行います。ぶっきらぼう心臓、下大静脈（IVC）、上大静脈（SVC）、上行大動脈、PA、および肺静脈を分離する。氷冷、滅菌生理食塩水でIVCを灌流。 </li><li> IVC、SVC、その後、より優れカット6​​-0絹縫合糸で肺静脈を結紮合字。 </li><li> 2ミリメートルカフと大動脈とPAをカット。 </li><li> PVを切り出し、それを処分する。 </li></ol> 3。心臓弁移植ドナー心臓の上に<ol><li>すぐにステップ2.5の後に、ドナー心臓にステップ1.5から心臓弁を配置し、心臓弁を向けます。 </li><li>テーパー針に10-0モノフィラメント縫合糸を使用したバルブの右側のステッチのPVを確保し、太陽光発電の反対側から5-6 stichesで連続縫合を開始します。 </li><li>前面を終えた後、水平に心を回転させ、ドナー心臓へのPVの裏側を縫合し始める。 </li><li>冷滅菌ヘパリン/食塩水に心を保管してください。注：ドナー心臓をレシピエントマウスへの移植前2時間溶液中に保持することができます。 </li></ol>受信者のマウスの上に4。異心臓移植<ol><li> 6-8週齢の雌C57BL / 6マウスRECIとして使用されたpient。そのケージからマウスを削除し、ケタミン/キシラジンカクテル（ケタミン、100 mg / kgのキシラジンおよび10 mg / kgの）で麻酔して、それを比較検討する。ケトプロフェン（5 mg / kgの）を、前麻酔鎮痛剤として使用される。 </li><li>つまんテールによる鎮静のレベルを確認した後、腹部と胸の毛をクリップ。無菌眼軟膏で目を潤滑し、パッド上の背側横臥位にマウスを置きます。ベタジンとアルコールパッドで腹部を消毒。その後、滅菌ドレープでマウスをカバーし、唯一の切開領域を公開します。 </li><li>恥骨領域への剣状突起下から正中開腹切開を行い、自己保持リトラクターを挿入します。生理食塩水湿らせたガーゼで腸を包む。ぶっきらぼう大動脈大動脈と大静脈を定義します。 </li><li>血液の循環を抑制するために大動脈と下大静脈を中心に近位および遠位2 6-0絹縫合糸を配置します。 </li><li>腹部大動脈の右側にドナー心臓を配置し、滅菌GAUでそれをカバーZE。生理食塩水で保湿し。 </li><li> 30 Gの針を用いて腹部大動脈に大動脈切開を行い、ドナーの大動脈の大きさにハサミで開口部を拡張する。 </li><li>テーパー針に無菌10-0モノフィラメント縫合糸を使用してエンドツー側吻合を行う。腹部大動脈の開口部の近位端に1ステッチとドナーの大動脈を確保し、腹部大動脈の先端から4-5 stichesで連続縫合を開始します。 </li><li> 、左側に心をフリップ生理食塩水を注入ガーゼでそれをカバーし、腹部大動脈の先端から4-5 stichesと連続して縫合糸に開始します。 </li><li> 30 Gの針を用いてIVCにおける静脈切開を行い、ドナー肺動脈の大きさに開口部を拡張する。 </li><li>テーパー針に無菌10-0モノフィラメント縫合糸を使用した側吻合に終止符を実行します。 IVCの開口部の近位端に1ステッチとドナーのPAを確保してから4-5 stichesで連続縫合を開始下大静脈の先端部。大動脈の方法であるため、今回は、ドナーのPAの左側壁の縫合は、IVCの内側にあることを確認してください。 </li><li>ヘパリンおよび生理食塩水（100単位/ ml）でIVC内腔を洗い流す。先端部に連続して、それらを縫合することにより、ドナーとレシピエントのPA IVCの右壁を閉じます。 </li><li>遠位合字を削除し、局所吸収性無菌止血剤を塗布して出血を制御します。出血が完全に停止すると、近位の縫合糸を取り除き、出血を同じように制御します。 </li><li>腸を返して6-0黒色ポリアミドモノフィラメント縫合糸を使用した2層に腹部の筋肉と皮膚を閉じます。 </li><li> 0.5ミリリットルの生理食塩水を皮下注射すると、マウスが完全にモバイルになるまで、温暖化パッド上で回復ケージにマウスを置きます。復旧時には、紙の寝具で新しいケージにマウスを返す。 48時間、鎮痛薬（イブプロフェン、30 mg / kgで、飲料水）を得た。やる完全に回復するまで、他の動物の会社に手術を受けている動物を返さない。 </li></ol>

<ol>
	<li>Polito, A., 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=Increased+morbidity+and+mortality+in+very+preterm+VLBW+infants+with+congenital+heart+disease.">Increased morbidity and mortality in very preterm VLBW infants with congenital heart disease.</a> Intens Care Med. 39, 1104-1112 (2013).</li><li>Wren, C., Reinhardt, Z., Khawaja, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Twenty+year+trends+in+diagnosis+of+life+threatening+neonatal+cardiovascular+malformations.">Twenty year trends in diagnosis of life threatening neonatal cardiovascular malformations.</a> Arch Dis Child Fetal. 93, F33-F35 (2008).</li><li>Vacanti, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beyond+transplantation+Third+annual+Samuel+Jason+Mixter+lecture.">Beyond transplantation Third annual Samuel Jason Mixter lecture.</a> Archives of surgery. 123, 545-549 (1960).</li><li>Tudorache, I., 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+replacement+of+aortic+heart+valves+with+tissue-engineered+grafts.">Orthotopic replacement of aortic heart valves with tissue-engineered grafts.</a> Tissue engineering Part A. 19, 1686-1694 (2013).</li><li>van Geldorp, M. 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=Patient+outcome+after+aortic+valve+replacement+with+a+mechanical+or+biological+prosthesis+weighing+lifetime+anticoagulant+related+event+risk+against+reoperation+risk.">Patient outcome after aortic valve replacement with a mechanical or biological prosthesis weighing lifetime anticoagulant related event risk against reoperation risk.</a> The Journal of thoracic and cardiovascular surgery. 137, 881-886 (2009).</li><li>El Oakley, R., Kleine, P., Bach, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Choice+of+prosthetic+heart+valve+in+today's+practice.">Choice of prosthetic heart valve in today's practice.</a> Circulation. 117, 253-256 (2008).</li><li>Quinn, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Models+for+Bench+to+Bedside+Translation+of+Regenerative+Cardiac+Constructs.">Animal Models for Bench to Bedside Translation of Regenerative Cardiac Constructs.</a> Progress in Pediatric cardiology. 35, (2013).</li><li>Shinoka, T., 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=Tissue+engineering+heart+valves+valve+leaflet+replacement+study+in+a+lamb+model.">Tissue engineering heart valves valve leaflet replacement study in a lamb model.</a> The Annals of thoracic surgery. 60, S513-S516 (1995).</li><li>Sodian, 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=Tissue+engineering+of+heart+valves+in+vitro+experiences.">Tissue engineering of heart valves in vitro experiences.</a> The Annals of thoracic surgery. 70, 140-144 (2000).</li><li>Shinoka, T., 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=Tissue+engineered+heart+valves.+Autologous+valve+leaflet+replacement+study+in+a+lamb+model.">Tissue engineered heart valves. Autologous valve leaflet replacement study in a lamb model.</a> Circulation. 94, II164-II168 (1996).</li><li>Sodian, 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=Evaluation+of+biodegradable+three+dimensional+matrices+for+tissue+engineering+of+heart+valves.">Evaluation of biodegradable three dimensional matrices for tissue engineering of heart valves.</a> ASAIO journal (American Society for Artificial Internal Organs. 46, 107-110 (1992).</li><li>Sodian, 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=Early+in+vivo+experience+with+tissue-engineered+trileaflet+heart+valves).">Early in vivo experience with tissue-engineered trileaflet heart valves).</a> Circulation. 102, III22-III29 (2000).</li></ol>

我々は、開示することは何もありません。

この手順の死亡率は、主に受信者に腹部大動脈へのドナーの大動脈でのPV移植部位および吻合での出血によって引き起こされた20パーセント、に近接しています。ほとんどの場合、死亡率が有意に48時間後に手術を減少させる。生存したマウスは、移植PVを介して強力な心拍、血流を示した。全体のプロセスは、経験豊富なマイクロ外科医4時間かかります。これは、技術を習得するためにおよ?...

先天性心血管の欠陥は西部の世界の1,2乳児死亡の主要な原因の1つです。その中でも、肺動脈弁狭窄症と二尖大動脈弁の欠陥が頻繁に生じる形態3である。心臓弁置換手術は、再建手術の通常の選択である。しかし、狭窄および石灰化心臓弁の、および抗凝固剤に終生の依存など、合併症は慢性的な病気と死4-7の重要な源である。また、成長の可能性がないことは、さらに、これらの若い患者4,8,9の死亡率を増加させる、リビジョン手術を必要とします。 生分解性合成心臓弁8上に成長性、Shinoka ら播種した自己由来細胞との機能的な置換心臓弁を開発する試みである。成長ポテンショ持つ構造体のようなネイティブの心臓弁に変換する合成バルブアル。予備大型動物研究は、機能的な心臓弁10を作成するために、この方法論を使用することの実現可能性を実証した。しかし、長期の移植研究は、心臓弁の狭窄を生じるバルブ新組織の進行性肥厚に乏しい耐久性を実証した。 Sodianから作業ら Shinoka手法を使用しましたが、最終的には、より生理的なプロフィール9,11,12を構築する組織工学弁の生体力学的特性を与えた生分解性エラストマーのPGA行列を置き換えました。 in vivo試験では、着床の成功にもかかわらず、コンフルエント内皮細胞ライニングが、この足場12の長期的な成功を制限する可能性が形成されなかった。 合理的に改善された第二世代合成心臓弁を設計するために、心臓弁移植のマウスモデルは、細胞および分子メカニズムを調べるために作成されたunderlyinGの新組織形成、バルブの肥厚、および狭窄の開発。マウスモデルは、他の種7で容易に入手できないトランスジェニックを含む分子試薬の広大な配列を提供しています。この心臓弁移植モデルにおいては、ex vivoで同系肺動脈心臓弁置換は、最初に実行した;した後、移植された心臓弁と心臓は顕微手法を用いて、同系宿主中に異所移植した。このモデルは、心肺バイパスを必要とせずに心臓弁置換を可能にします。 本研究では、心臓弁の収穫の詳細な説明は、ドナー心臓製剤、心臓弁移植、および異所心臓移植について説明する。結果は、受信者のハートビートとは無関係であったドナー心臓から連続ハートビートを示した。移植された肺動脈弁を通る血流がパルス波飲酒して高周波数超音波システムを用いて測定したppler。

<table><tbody><tr><td>DPBS</td><td>Gibco</td><td>14190-144</td><td></td></tr><tr><td>Microscope</td><td>Leica</td><td>M80</td><td></td></tr><tr><td>C57BL/6J (H-2b), Female</td><td>Jackson Laboratories</td><td>664</td><td>8-12 weeks</td></tr><tr><td>Ketamine Hydrochloride Injection</td><td>Hospira Inc.</td><td>NDC 0409-2053</td><td></td></tr><tr><td>Xylazine Sterile Solution</td><td>Akorn Inc.</td><td>NADA# 139-236</td><td></td></tr><tr><td>Ketoprofen</td><td>Fort Dodge Animal Health</td><td>NDC 0856-4396-01</td><td></td></tr><tr><td>Ibuprofen</td><td>PrecisionDose</td><td>NDC 68094-494-59</td><td></td></tr><tr><td>Heparin Sodium</td><td>Sagent Pharmaceticals</td><td>NDC 25021-400</td><td></td></tr><tr><td>Saline solution (Sterile 0.9% sodium chloride)</td><td>Hospira Inc.</td><td>NDC 0409-0138-22</td><td></td></tr><tr><td>0.9% Sodium chloride Injection</td><td>Hospira Inc.</td><td>NDC 0409-4888-10</td><td></td></tr><tr><td>Petrolatum Ophthalmic Ointment</td><td>Dechra Veterinary Products</td><td>NDC 17033-211-38</td><td></td></tr><tr><td>Iodine Prep Pads</td><td>Triad Disposables, Inc.</td><td>NDC 50730-3201-1</td><td></td></tr><tr><td>Alcohol Prep Pads</td><td>McKesson Corp.</td><td>NDC 68599-5805-1</td><td></td></tr><tr><td>Cotton tipped applicators</td><td>Fisher Sientific</td><td>23-400-118</td><td></td></tr><tr><td>Fine Scissor</td><td>FST</td><td>14028-10</td><td></td></tr><tr><td>Micro-Adson Forcep</td><td>FST</td><td>11018-12</td><td></td></tr><tr><td>Clamp Applying Forcep</td><td>FST</td><td>00072-14</td><td></td></tr><tr><td>S&amp;amp;T Vascular Clamp</td><td>FST</td><td>00396-01</td><td></td></tr><tr><td>Spring Scissors</td><td>FST</td><td>15008-08</td><td></td></tr><tr><td>Colibri Retractors</td><td>FST</td><td>17000-04</td><td></td></tr><tr><td>Dumont #5 Forcep</td><td>FST</td><td>11251-20</td><td></td></tr><tr><td>Dumont #7 - Fine Forceps</td><td>FST</td><td>11274-20</td><td></td></tr><tr><td>Dumont #5/45 Forceps</td><td>FST</td><td>11251-35</td><td></td></tr><tr><td>Tish Needle Holder/Forceps</td><td>Micrins</td><td>MI1540</td><td></td></tr><tr><td>Black Polyamide Monofilament Suture, 10-0</td><td>AROSurgical Instruments Corporation</td><td>TI638402</td><td>For sutureing the graft</td></tr><tr><td>Black Polyamide Monofilament Suture, 6-0</td><td>AROSurgical Instruments</td><td>SN-1956</td><td>For musculature and skin closure</td></tr><tr><td>Non Woven Sponges</td><td>McKesson Corp.</td><td>94442000</td><td></td></tr><tr><td>Absorbable hemostat</td><td>Ethicon</td><td>1961</td><td></td></tr><tr><td>1 ml Syringe</td><td>BD</td><td>309659</td><td></td></tr><tr><td>3 ml Syringe</td><td>BD</td><td>309657</td><td></td></tr><tr><td>10 ml Syringe</td><td>BD</td><td>309604</td><td></td></tr><tr><td>18 G 1 1/2 in, Needle</td><td>BD</td><td>305190</td><td></td></tr><tr><td>25 G 1 in., Needle</td><td>BD</td><td>305125</td><td></td></tr><tr><td>30 G 1 in., Needle</td><td>BD</td><td>305106</td><td></td></tr><tr><td>Warm Water Recircultor</td><td>Gaymar</td><td>TP-700</td><td></td></tr><tr><td>Warming Pad</td><td>Gaymar</td><td>TP-22G</td><td></td></tr><tr><td>Trimmer</td><td>Wahl</td><td>9854-500</td><td></td></tr><tr><td>VEVO2100 High Frequency Ultrasound</td><td>VisualSonics</td><td>&lt;a href=&quot;http://www.visualsonics.com/vevo2100&quot;&gt;http://www.visualsonics.com/vevo2100&lt;/a&gt;</td><td>The catalog number and pricing can be acquired from the sales representatives.</td></tr><tr><td>Ultrasound transmission gel</td><td>Parker Laboratories,&lt;br/&gt; INC.</td><td>01-02</td><td></td></tr><tr><td>Table Top Laboratory Animal Anesthesia System</td><td>VetEquip, INC.</td><td>901806</td><td></td></tr><tr><td>Isoflurane</td><td>Baxter</td><td>1001936060</td><td></td></tr></tbody></table>

transplantation of pulmonary valve using a mouse model of heterotopic heart transplantation

組織工学心臓弁、特に脱細胞弁は、まちまちな結果と再建手術の臨床使用で躍進を始めている。しかし、新組織開発、弁の肥厚および狭窄の発生の細胞および分子メカニズムは広く研究されていない。上記の質問に答えるために、我々は、マウス異心臓弁移植モデルを開発しました。心臓弁は、弁ドナーマウスから採取し、心臓ドナーマウスに移植した。新しいバルブに心臓がレシピエントマウスに異所移植した。移植された心臓は、受信者の心拍とは無関係に、独自のビートを示した。血流は、パルス波ドプラで高周波数超音波システムを用いて定量した。移植された肺動脈弁を通る流れは、最小限の逆流との流れを前進示し、ピークフローは100mm /秒に近いものであった。心臓弁移植のこのマウスモデルであるhighl多用途のyので、変性及び異なる血行力学環境​​を提供するために、および/または組織工学心臓弁に新組織発達を研究するために、種々のトランスジェニックマウスと共に使用することができるように適合させることができる。

図1は、異所性心臓移植を用いた心臓弁移植モデルの概略図を示す。心臓弁は、ドナー心臓から採取し、第二のドナーマウスから心臓に移植した。次に、新しい心臓弁と心臓はレシピエントマウスの腹部に移植した。 図2は、腹部の空間（A）、右心臓移植（B）の後、および移植後5分に注入された心臓の図を示している。大動脈と下大静脈の?...

Watch this Scientific Journal Video about Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation at JoVE.com

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

組織工学心臓弁における新組織形成および狭窄の開発の基礎となる細胞および分子メカニズムを理解するためには、異所性心臓弁移植のマウスモデルを開発した。肺動脈心臓弁は異所心臓移植技術を用いてレシピエントに移植した。

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 ...

transplantation-pulmonary-valve-using-mouse-model-heterotopic-heart

Research

JoVE Journal

Medicine

18.8K ビュー.  Nationwide Children's Hospital. 組織工学心臓弁における新組織形成および狭窄の開発の基礎となる細胞および分子メカニズムを理解するためには、異所性心臓弁移植のマウスモデルを開発した。肺動脈心臓弁は異所心臓移植技術を用いてレシピエントに移植した。 

ビデオ: Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

Heterotopic Heart Transplantation in Mice

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable for studying rejection and immune response now that newer transgenic and gene knockout mice are available, and a large number of immunologic reagents have been developed. The heart transplant model is less stringent than the skin transplant models, although technically more challenging. We have developed a modified technique and have completed over 1000 successful cases of heterotopic heart transplantation in mice. When making anastomosis of the ascending aorta and abdominal aorta, two stay sutures are placed at the proximal and distal apexes of recipient abdominal aorta with the donor s ascending aorta, then using 11-0 suture for anastomosis on both side of aorta with continuing sutures. The stay sutures make the anastomosis easier and 11-0 is an ideal suture size to avoid bleeding and thrombosis.
When making anastomosis of pulmonary artery and inferior vena cava, two stay sutures are made at the proximal apex and distal apex of the recipient s inferior vena cava with the donor s pulmonary artery. The left wall of the inferior vena cava and donor s pulmonary artery is closed with continuing sutures in the inside of the inferior vena cava after, one knot with the proximal apex stay suture the right wall of the inferior vena cava and the donor s pulmonary artery are closed with continuing sutures outside the inferior vena cave with 10-0 sutures. This method is easier to perform because anastomosis is made just on the one side of the inferior vena cava and 10-0 sutures is the right size to avoid bleeding and thrombosis. In this article, we provide details of the technique to supplement the video.

The mouse heterotopic heart transplantation model has been proven by many investigators to be an important method for studying mechanisms of rejection and immune response. However, the techniques involved are still challenging. By modifying standard techniques we have had success with more than 1000 transplants.

マウスにおける異所心臓移植

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable ...

生物学

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 ...

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 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 ...

Mouse Heterotopic Cervical Cardiac Transplantation Utilizing Vascular Cuffs

Murine models of cardiac transplantation are frequently utilized to study ischemia-reperfusion injury, innate and adaptive immune responses after transplantation, and the impact of immunomodulatory therapies on graft rejection. Heterotopic cervical heart transplantation in mice was first described in 1991 using sutured anastomoses and subsequently modified to include cuff techniques. This modification allowed for improved success rates, and since then, there have been multiple reports that have proposed further technical improvements. However, translation into more widespread utilization remains limited due to the technical difficulty associated with graft anastomoses, which requires precision to achieve adequate length and caliber of the cuffs to avoid vascular anastomotic twisting or excessive tension, which can result in damage to the graft. The present protocol describes a modified technique for performing heterotopic cervical cardiac transplantation in mice which involves cuff placement on the recipient's common carotid artery and the donor's pulmonary artery in alignment with the direction of the blood flow.

Mouse cardiac transplantation models represent valuable research tools for studying transplantation immunology. The present protocol details mouse heterotopic cervical cardiac transplantation that involves the placement of cuffs on the recipient's common carotid artery and the donor's pulmonary artery trunk to allow for laminar blood flow.

血管カフを用いたマウスヘテロトピック頸部心臓移植

Murine models of cardiac transplantation are frequently utilized to study ischemia-reperfusion injury, innate and adaptive immune responses after ...

免疫・感染学

Blood Circuit Reconstruction in an Abdominal Mouse Heart Transplantation Model

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the established technique for a modified blood circuit reconstruction in a heterotopic abdominal heart transplantation model is presented. This method uses the intrathoracic inferior vena cava (IIVC) instead of the pulmonary artery of the donor heart for the anastomosis to the inferior vena cava of the recipient. It is facilitating and improving success rates for abdominal heart transplantation in mice.

A novel technique for blood circuit reconstruction in a heterotopic abdominal mouse heart transplantation model is demonstrated.

腹部マウス心臓移植モデルにおける血液回路再建

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the ...

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 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 ...