この作品は賞号W81XWH-13-2-0053の下で、AFIRM II努力をサポートするために、陸軍、海軍、NIH、空軍、VAおよび保健省によってサポートされていました。米陸軍医学研究買収活動、820チャンドラーストリート、フォート・デトリックMD 21702から5014までは表彰および管理取得オフィスです。ご意見、解釈、結論と勧告は著者のものであり、必ずしも国防総省によって承認されていません。 著者らは、この研究の間に、その優れた獣医のサポートのためにジェシカイッチ、DVM、キャロライン・ギャレット、DVMとジュリー・ワトソン、DVMに感謝したいと思います。

全ての実験は、国立衛生研究所の実験動物の管理と使用に関する指針（NIH）に従って行ったし、ジョンズ・ホプキンス大学動物実験委員会（JHUACUC）によって承認されました。具体的な手順は、承認されたACUCプロトコルMO13M108下で行いました。 1.ドナー操作<ol><li>手術前に各薬理学的製剤のための適切な時点で鎮痛を管理します。承認された動物の世話のとおりと前皮膚切開にブプレノルフィン皮下に1時間の0.1ミリグラム/キログラムBWプロトコルの使用を使用します。 </li><li>イソフルランで落ち着いたドナーは4％でイソフルラン気化器に取り付けられた室を通って適用しました。ノーズコーンを介して2％で鎮静や麻酔を維持します。手続きの開始に先立って麻酔深度を監視するためにつま先ピンチ撤退反射を実行します。 </li><li>マスク、使い捨ての分離ガウンと手袋を着用してください。 </li><li>外科的ARを剃りますEA、特に後肢と鼠径部、および10％のポビドンで準備を - ヨウ素。 </li><li>滅菌野ドレープ、オートクレーブ処理機器や高倍率顕微鏡（40X）を使用します。 </li><li>半ば腿領域に近位にはさみを使用して、脚の付け根の皮膚切開を行い、円周方向にマウス本体の残りの部分から後肢を画定するために切開部を接続します。 </li><li>大腿動脈、静脈や神経を特定し、解剖。鉗子とマイクロはさみを使用して、すべての3つの構造を分離します。 </li><li>血管茎を解剖したら、マイクロはさみを使用して、鼠径靱帯のレベルで血管を分割します。 </li><li>次に、はさみを使用して、ドナー動物からの移植片を分離するために半ば腿のレベルで近位個々の腹側（薄と大腿内側の筋肉）と背側の筋肉群20を分割し続けます。 </li><li>大腿骨を横断し、はさみを使用して大腿骨シャフトの中間で切断。 </li><li>イソフルラン過剰摂取FOLにより動物を安楽死させます頚椎脱臼が続き。心拍や呼吸の停止を確認してください。 </li><li>注射器に取り付けられた33のGフラッシング針を使用してヘパリン化2ミリリットル（30 IE）の冷（4℃）生理食塩水で手足をフラッシュ（材料表を参照してください）​​。 </li><li>それぞれ、大腿静脈と動脈に1ポリイミドカフを配置します。 </li><li>インセットまで湿った綿ガーゼ、4℃でペトリ皿や店舗内の所定の場所に移植片をラップします。 </li></ol> 2.受信者の操作<ol><li> 後肢の除去 <ol><li>手術前に各薬理学的製剤のための適切な時点で鎮痛を管理します。承認された動物の管理と使用プロトコルを使用ブプレノルフィンSC 1時間前に皮膚切開の0.1ミリグラム/ kg体重あたりとして。 </li><li>イソフルランで落ち着いたドナーは4％でイソフルラン気化器に取り付けられた室を通って適用しました。ノーズコーンを介して2％で鎮静や麻酔を維持します。 DEPTを監視するために、つま先のピンチ撤退の反射を行います手続きの開始に先立って麻酔の時間。 </li><li>麻酔下ながら乾燥を防ぐために、マウスの目に獣医の軟膏を使用してください。 </li><li>特に、10％ポビドンと後肢と脚の付け根と準備を手術領域を剃る - ヨウ素。 </li><li>半ば腿領域に近位にはさみを使用して、脚の付け根の皮膚切開を行い、円周方向にマウス本体の残りの部分から後肢を画定するために切開部を接続します。 </li><li>鉗子とマイクロはさみを使用して、すべての3つの構造を特定し、大腿動脈、静脈や神経を解剖し、分離します。 </li><li>血管茎を解剖したら、鼠径靱帯のレベルで大腿血管をクランプします。 </li><li>浅腹壁動脈のレベルで遠位血管をカットします。 </li><li>次に、ネイティブ後肢リチウムを分離するために、近位に半ば腿のレベルで個々の腹側（薄と大腿内側の筋肉）と背側の筋肉群20を分割し続けますはさみを使用して、レシピエント動物からMB。 </li><li>はさみを使用して大腿骨シャフトの中央に大腿骨を横断。 </li><li>切開部位の出血、したがってレシピエントの血液損失を防止するために、以前に離断大腿部の筋肉を焼灼。 </li></ol></li><li> 移植 <ol><li>温かい生理食塩水（37°C）で術野を灌漑し、操作の前と後の0.3ミリリットル暖かい生理食塩水を注入することにより、流体の損失を最小限に抑えます。 </li><li>受信者の大腿骨と移植片を整列させることによって、ネイティブ後肢の正確な解剖学的位置を反映し、髄内ロッドとして20 G脊髄針を使用してそれらを接続する方法で、移植片を置きます。 </li><li>吸収性縫合糸材料（6-0ポリソーブ）を使用して癒合腹側と背側の筋肉群。 </li><li>非縫合カフ技術を用いて大腿血管を接続します。詳細に、以前グラーフの容器端部に取り付けられたカフの上に容器の受信者側を引っ張りますトン。 10-0ナイロン縫合糸を使用し、袖口に受信者の容器を固定するために、円周ネクタイを行います。 </li><li>次のクランプを解放します。この段階で視覚的にカフ回転および血管の誤回転やねじれを防止するための最適な位置を確認します。 </li><li>筋肉の受信者ドナーインターフェースと骨の両端を中心とした電気メスを使用して細心の止血を行います。 </li><li>非吸収性ナイロン縫合糸（6-0 Ethilon）を用いて皮膚を閉じます。 </li><li>動物は加熱ランプの下で、そのケージに回復できるようにすることで、正常体温条件を確立します。住宅施設に戻す前に、少なくとも4時間、定期的なモニタリングを続けます。 </li><li> 3日間は0.1mg / kgをSC毎に6-8時間の用量でブプレノルフィンと術後鎮痛を提供します。 </li></ol></li></ol>

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Med. 368, 1850-1852 (2013).</li><li>Leventhal, J., 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=Chimerism+and+tolerance+without+GVHD+or+engraftment+syndrome+in+HLA-mismatched+combined+kidney+and+hematopoietic+stem+cell+transplantation.">Chimerism and tolerance without GVHD or engraftment syndrome in HLA-mismatched combined kidney and hematopoietic stem cell transplantation.</a> Sci. Transl. Med. 4, 124-128 (2012).</li><li>Cendales, L. C., 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+Banff+2007+working+classification+of+skin-containing+composite+tissue+allograft+pathology.">The Banff 2007 working classification of skin-containing composite tissue allograft pathology.</a> Am. J. Transplant. 8, 1396-1400 (2008).</li></ol>

The authors declare that they have no competing financial interest.

このような壊滅的な組織欠損の再建のための上肢と顔面移植などの血管新生したコンポジット同種移植は、従来の再建手続きに修正可能ではない患者に対する有効な治療法の選択肢として発展してきました。マイクロサージャリーの分野における技術の進歩だけでなく、強力な免疫抑制および固形臓器移植における免疫調節療法と豊富な経験は、今、このユニークな患者集団3,21の長?...

The late nineties heralded the pioneering days of reconstructive transplantation with the first successful hand transplant performed in France in 1998. Since then, the use of VCAs for reconstruction of devastating tissue defects has been successfully employed in a wide spectrum of patients. To date, the world counts 76 recipients of 112 upper extremities as well as 31 faces 1-3. In addition, several other types of VCAs such as abdominal wall 4, larynx 5, trachea 6, vascularized joints 7, and even penis 8 have been performed. Furthermore, the live birth of a baby was recently reported after uterus transplantation 9. This growing world experience is indicative for how reconstructive transplantation has become a valid therapeutic option for patients suffering of significant functional tissue defects not amendable to conventional reconstructive and restorative surgery and treatment. 
While the idea of replacing "like with like" sparked clinical enthusiasm, initial skepticism still prevails with regards to side effects of conventional high-dose immunosuppression required to maintain allografts and their function 10,11. However, as shown by seminal work of Lee et al., these composite grafts are less likely to reject than its individual components, and furthermore, some of the tissue components such as the vascularized bone compartment have fueled optimism as they might exert unique immunological effects onto the balance of immune acceptance and rejection 12. 
Our group pioneered several microsurgical animal models for solid organ transplantation, as well as vascularized composite allotransplantation 13-19. Here we describe a novel surgical procedure using a non-suture cuff technique to perform super micro-vascular anastomosis in an orthotopic mouse hind limb transplantation model. This transplant model provides a useful tool for investigating immune acceptance and rejection mechanisms, as well as the role of individual tissue components, such as the vascularized bone marrow compartment, towards tolerance induction in the immunologically versatile setting of the mouse species. Additionally, the orthotopic placement of the limb opens the possibilities for nerve regeneration and functional outcome studies, which are critically important to the setting of VCA.

<table><tbody><tr><td>Suture, 6-0 Nylon</td><td>MWI</td><td>31849</td><td></td></tr><tr><td>Suture, 6-0 Polysorb</td><td>MWI</td><td>72667</td><td></td></tr><tr><td>Suture, 10-0 Nylon</td><td>Aero Surgical</td><td>TK-107038</td><td></td></tr><tr><td>Polyimide Tubing, Size 25</td><td>Vention Medical</td><td>141-0023</td><td></td></tr><tr><td>Polyimide Tubing, Size 27</td><td>Vention Medical</td><td>141-0015</td><td></td></tr><tr><td>Microvascular Clamps (Single)</td><td>Synovis</td><td>00396</td><td></td></tr><tr><td>Microvascular Clamps (Double)</td><td>Synovis</td><td>00414</td><td></td></tr><tr><td>Micro-Scissors</td><td>Synovis</td><td>SAS-18</td><td></td></tr><tr><td>Micro-Forceps</td><td>Synovis</td><td>FRS-15 RM-8</td><td></td></tr><tr><td>Micro-Dilators</td><td>Synovis</td><td>FRS-15 RM-8d.1</td><td></td></tr><tr><td>Micro-Needledriver</td><td>Synovis</td><td>C-14</td><td></td></tr><tr><td>Micro-Clamp Applicator</td><td>Synovis</td><td>CAF-4</td><td></td></tr><tr><td>Micro-Flushing Needle</td><td>Hamilton</td><td></td><td>10MM, 30&amp;deg;, 33G</td></tr><tr><td>Lactated Ringers Solution</td><td>Fisher Scientific</td><td>NC9968051</td><td></td></tr><tr><td>Buprenorphine</td><td></td><td></td><td>DEA Number required; Obtained from hosptial pharmacy.</td></tr><tr><td>Enrofloxacin; Baytril</td><td>Bayer Health Care</td><td>186599</td><td></td></tr><tr><td>Heparin</td><td></td><td></td><td>Obtained from hosptial pharmacy</td></tr></tbody></table>

orthotopic hind limb transplantation in the mouse

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and translational research in the field of transplantation medicine. A vast array of reagents is available exclusively in this setting, including mono- and polyclonal antibodies for both diagnostic and interventional applications. In addition, a vast number of genotyped, inbred, transgenic, and knock out strains allow detailed investigation of the individual contributions of humoral and cellular components to the complex interplay of an immune response and make the mouse the gold standard for immunological research. 
Vascularized Composite Allotransplantation (VCA) delineates a novel field of transplantation using allografts to replace "like with like" in patients suffering traumatic or congenital tissue loss. This surgical methodological protocol shows the use of a non-suture cuff technique for super-microvascular anastomosis in an orthotopic mouse hind limb transplantation model. The model specifically allows for comparison between established paradigms in solid organ transplantation with a novel form of transplants consisting of various different tissue components. Uniquely, this model allows for the transplantation of a viable vascularized bone marrow compartment and niche that have the potential to exert a beneficial effect on the balance of immune acceptance and rejection. This technique provides a tool to investigate alloantigen recognition and allograft rejection and acceptance, as well as enables the pursuit of functional nerve regeneration studies to further advance this novel field of transplantation.

非縫合カフ技術を用いて、マウスモデルにおいて血管新生複合同種移植を行うと、 図1に示すように優れ、長期間の移植および動物の生存を達成することができる。また、血管新生合成の緩やかな同種移植片拒絶反応の再現可能な結果を得るための信頼できる方法を表します。などの同種移植は、 図2に示されている画像によって文書化。拒?...

Watch this Scientific Journal Video about Orthotopic Hind Limb Transplantation in the Mouse at JoVE.com

Orthotopic Hind Limb Transplantation in the Mouse

超微小血管吻合のための非縫合カフ技術を適用したマウスにおける同所後肢移植のためのこの新規モデルは、血管新生複合同種移植（VCA）に関連したin vivoでのメカニズムの免疫学的研究のための強力なツールを提供します。

マウスで同所後肢移植

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and ...

orthotopic-hind-limb-transplantation-in-the-mouse

Research

JoVE Journal

Medicine

11.6K ビュー.  Johns Hopkins University School of Medicine. 超微小血管吻合のための非縫合カフ技術を適用したマウスにおける同所後肢移植のためのこの新規モデルは、血管新生複合同種移植（VCA）に関連したin vivoでのメカニズムの免疫学的研究のための強力なツールを提供します。 

ビデオ: Orthotopic Hind Limb Transplantation in the Mouse

Orthotopic Hind-Limb Transplantation in Rats

Composite tissue allotransplantation (CTA) now represents a valid therapeutic option after the loss of a hand, forearm or digits and has become a novel therapeutic entity in reconstructive surgery. However, long term high-dose multi-drug immunosuppressive therapy is required to ensure graft survival, bearing the risk of serious side effects which halters broader application. Further progression in this field may depend on better understanding of basic immunology and ischemia reperfusion injury in composite tissue grafts.
To date, orthotopic hind limb transplantation in rats has been the preferred rodent model for reconstructive transplantation (RT), however, it is an extremely demanding procedure that requires extraordinary microsurgical skills for reattachment of vasculature, bones, muscles and nerves.
We have introduced the vascular cuff anastomosis technique to this model, providing a rapid and reliable approach to rat hind limb transplantation. This technique simplifies and shortens the surgical procedure and enables surgeons with basic microsurgical experience to successfully perform the operation with high survival and low complication rates. The technique seems to be well suited for immunological as well as ischemia reperfusion injury (IRI) studies.

Here we describe the orthotopic rat hind-limb transplantation procedure, which seems to be the gold standard in vivo model for composite tissue allotransplantation research.

ラット同所性後肢移植

Composite tissue allotransplantation (CTA) now represents a valid therapeutic option after the loss of a hand, forearm or digits and has become a novel ...

免疫・感染学

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

Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. This is largely true, but there are a number of important principles that can reduce the difficulty of these procedures and enhance reproducibility. Orthotopic aortic transplantation is an excellent procedure in which to learn these principles because it involves only two end-to-end anastomoses, but requires good suturing technique and handling of the vessels for consistent success. This procedure begins with the procurement of a length of abdominal aorta from a donor animal, followed by division of the native aorta in the recipient. The procured aorta is then placed between the divided ends of the recipient aorta and sutured into place using end-to-end anastomoses. To accomplish this objective successfully requires a high degree of concentration, good tools, a steady hand, and an appreciation of how easily the vasculature of a mouse can be damaged, resulting in thrombosis. Learning these important principles is what occupies most of the beginner's time when learning microsurgery in small rodents. Throughout this protocol, we refer to these important points. This model can be used to study vascular disease in a variety of different experimental systems1-8. In the context shown here, it is most often used for the study of post-transplant vascular disease, a common long-term complication of solid organ transplantation in which intimal hyperplasia occurs within the allograft. The primary advantage of the model is that it facilitates quantitative morphometric analyses and the transplanted vessel lies contiguous to the endogenous vessel, which can serve as an additional control9. The technique shown here is most often used for mice weighing 18-25 grams. We have accumulated most of our experience using the C57BL/6J, BALB/cJ, and C3H/HeJ strains.

We describe a technique in which a section of the abdominal aorta from a mouse is transplanted orthotopically to just below the renal arteries in an allogeneic or syngeneic recipient. This technique can be useful in studies in which transplantation of large arteries of uniform size is deemed advantageous.

血管疾患の研究のためのマウスにおける同所性大動脈移植

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. ...

医学

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 ...

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 ...

Partial Heterotopic Hindlimb Transplantation Model in Rats

Vascularized composite allotransplantations (VCA) represent the most advanced reconstruction option for patients without autologous surgical possibilities after a complex tissue defect. Face and hand transplantations have changed disfigured patients' lives, giving them a new aesthetic and functional social organ. Despite promising outcomes, VCA is still underperformed due to life-long immunosuppression comorbidities and infectious complications. The rat is an ideal animal model for in vivo studies investigating immunological pathways and graft rejection mechanisms. Rats are also widely used in novel composite tissue graft preservation techniques, including perfusion and cryopreservation studies. Models used for VCA in rats must be reproducible, reliable, and efficient with low postoperative morbidity and mortality. Heterotopic limb transplantation procedures fulfill these criteria and are easier to perform than orthotopic limb transplants. Mastering rodent microsurgical models requires solid experience in microsurgery and animal care. Herein is reported a reliable and reproducible model of partial heterotopic osteomyocutaneous flap transplantation in rats, the postoperative outcomes, and the means of prevention of potential complications.

This paper presents a partial heterotopic osteomyocutaneous flap transplantation protocol in rats and its potential outcomes in the mid-term follow-up.

ラットにおける部分的な異所性後肢移植モデル

Vascularized composite allotransplantations (VCA) represent the most advanced reconstruction option for patients without autologous surgical possibilities ...

A Modified Murine Heterotopic Heart Transplant Protocol Matching Contemporary Standards of Aseptic Technique, Anesthesia, and Analgesia

The development of experimental models of cardiac transplantation in animals has contributed to many advances in the fields of immunology and solid organ transplantation. While the heterotopic vascularized murine cardiac transplantation model was initially utilized in studies of graft rejection using combinations of mismatched inbred mouse strains, access to genetically modified strains and therapeutic modalities can provide powerful new preclinical insights. Fundamentally, the surgical methodology for this technique has not changed since its development, especially with respect to important factors such as aseptic technique, anesthesia, and analgesia, which make material impacts on postsurgical morbidity and mortality. Additionally, improvements in perioperative management are expected to provide improvements in both animal welfare and experimental outcomes. This paper reports upon a protocol developed in collaboration with a subject matter expert in veterinary anesthesia and describes the surgical technique with an emphasis on perioperative management. Additionally, we discuss the implications of these refinements and provide details on troubleshooting critical surgical steps for this procedure.

The present paper describes a modified technique for heterotopic vascularized cardiac transplantation with updated aseptic technique, analgesia, and anesthesia.

無菌技術、麻酔、および鎮痛の現代の基準に一致する修正マウス異所性心臓移植プロトコル

The development of experimental models of cardiac transplantation in animals has contributed to many advances in the fields of immunology and solid organ ...

Taking the Next Step: a Neural Coaptation Orthotopic Hind Limb Transplant Model to Maximize Functional Recovery in Rat

Limb transplant in particular and vascularized composite allotransplant (VCA) in general have wide therapeutic promise that have been stymied by current limitations in immunosuppression and functional neuromotor recovery. Many animal models have been developed for studying unique features of VCA, but here we present a robust reproducible model of orthotopic hind limb transplant in rats designed to simultaneously investigate both aspects of current VCA limitation: immunosuppression strategies and functional neuromotor recovery. At the core of the model rests a commitment to meticulous, time-tested microsurgical techniques such as hand sewn vascular anastomoses and hand sewn neural coaptation of the femoral nerve and the sciatic nerve. This approach yields durable limb reconstructions that allow for longer lived animals capable of rehabilitation, resumption of daily activities, and functional testing. With short-term treatment of conventional immunosuppressive agents, allotransplanted animals survived up to 70 days post-transplant, and isotransplanted animals provide long lived controls beyond 200 days post-operatively. Evidence of neurologic functional recovery is present by 30 days post operatively. This model not only provides a useful platform for interrogating immunological questions unique to VCA and nerve regeneration, but also allows for in vivo testing of new therapeutic strategies specifically tailored for VCA.

This protocol presents a robust, reproducible model of vascularized composite allotransplant (VCA) geared toward simultaneous study of immunology and functional recovery. The time invested in meticulous technique in a right mid-thigh hind limb orthotopic transplant with hand sewn vascular anastomoses and neural coaptation yields the ability to study functional recovery.

次のステップを踏む:ラットの機能回復を最大化するための神経凝集整体交位後肢移植モデル

Limb transplant in particular and vascularized composite allotransplant (VCA) in general have wide therapeutic promise that have been stymied by current ...

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 Heterotopic Mouse Model for Studying Laryngeal Transplantation

Laryngeal heterotopic transplantation, although a technically challenging procedure, offers more scientific analysis and cost benefits compared to other animal models. Although first described by Shipchandler et al. in 2009, this technique is not widely used, possibly due to the difficulties in learning the microsurgical technique and time required to master it. This paper describes the surgical steps in detail, as well as potential pitfalls to avoid, in order to encourage effective use of this technique.
In this model, the bilateral carotid arteries of the donor larynx are anastomosed to the recipient carotid artery and external jugular vein, allowing for blood flow through the graft. Blood flow can be confirmed intraoperatively by the visualization of blood filling in the graft bilateral carotid arteries, reddening of the thyroid glands of the graft, and bleeding from micro vessels in the graft. The crucial elements for success include delicate preservation of the graft vessels, making the correct size arteriotomy and venotomy, and using the appropriate number of sutures on the arterial-arterial and arterial-venous anastomoses to secure vessels without leakage and prevent occlusion. 
Anyone can become proficient in this model with sufficient training and perform the procedure in approximately 3 h. If performed successfully, this model allows for immunologic studies to be performed with ease and at low cost.

The aim of this manuscript is to describe the microsurgical steps required to perform a heterotopic laryngeal transplant in mice. The advantages of this mouse model compared to other animal models of laryngeal transplantation are its cost-effectiveness and the availability of immunologic assays and data.

喉頭移植研究のための異所性マウスモデル

Laryngeal heterotopic transplantation, although a technically challenging procedure, offers more scientific analysis and cost benefits compared to other ...