Name: a rat model of orthotopic liver transplantation using a novel magnetic anastomosis technique for suprahepatic vena cava reconstruction
Uploaded: 2018-03-19T12:30:00.000+00:00
Duration: 14 min 33 s
Description: Watch this Scientific Journal Video about A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction at JoVE.com

この作品は、省の教育革新チーム開発プログラム中国の号からの補助金によって支えられました。IRT16R57)、中国 (第 81470896)、研究の国家自然科学基金の若い才能を募集予定の西安交通大学 (RW) に資金を提供します。

プロトコルは、ケアと実験動物の使用のためのガイドラインに従って行われた、実験動物の倫理の西安交通大学、西安、陝西省、中国の委員会によって承認されました。
注: このプロトコルの手順のどれもを手術用顕微鏡の下で行った。
1. 術前準備
<ol><li>デザインと形状の磁気リング (図 1) の<ol>
<li>ネオジム-鉄-ホウ素磁気リングのペアを照合することによって磁気吻合デバイスを組み立てます。</li>
		<li>それぞれ主要な軸径、短軸径、厚さ 8 mm、5.5 mm、1 mm の 0.5 g の重量とラット グラフトのパラメーターによるとリングをデザインします。</li>
		<li>ワイヤ電極切削法による曲線の所望の形状にリング チタン酸化物エアブラシで得られたリングをコートし、500 mg/L 6 h の 55-60 ° C でエチレン酸化物と消毒します。</li>
	</ol>
</li>
	<li>カフス (図 2)<ol>
<li>ポリエチレン管を切断することによって IHVC と PV の吻合のカフを準備します。確実に 5 mm カフ体 2 mm カフ拡張子と長い。 注: IHVC カフのそれらは、2.6 と 2.8 mm、それぞれありますが、PV カフの内径、外径、1.8 と 2.1 mm です。</li>
	</ol>
</li>
	<li>ステント (図 2)<ol>
<li>両端に傾斜を生成する長さ 5 mm の切削 24 G 静脈カニューレによって BD の吻合のためのステントを準備します。</li>
	</ol>
</li>
	<li>実験動物<ol>
<li>ドナーとして 250 280 g と受信者との間の重量を量る男性スプレイグ ラットを使用します。ドナーよりも、受信者が同じ重量またはわずかに重い (< 10 g) であることを確認します。</li>
		<li>食料と水は手術前に回線付き客室の室温制御にラットをしてください。</li>
	</ol>
</li>
</ol>2. ドナー操作
<ol><li>注入ブプレノルフィン (0.05 mg/kg 皮下鎮痛剤として操作の前に 1 時間。</li>
	<li>イソフルラン吸入によるドナー ラットを麻酔します。5 vol % イソフルラン麻酔および流量 0.6 0.8 で 2 vol % イソフルランの誘導のプレキシ グラス箱に 1.5 L/分の流量で L/円錐分マスク麻酔の維持のため。つま先のピンチと皮膚のピンチを実行することによって麻酔の深さを確認します。</li>
	<li>クリーナーの露出を許可する電気シェーバーを使用してラットの腹部の肌全体を剃る。ポビドン ヨード液と対応する皮膚を消毒します。</li>
	<li>に沿って部の 3部上 1 cm 拡張するペニスの根本から手術用のはさみを使用して十字切開を介して腹腔内を開くlinea alba、腋窩中線に縦切開の中点から延びる。左に消化管を外在し、湿らせたガーゼでカバーします。</li>
	<li>クランプ、止血鉗子を用いた頭に向かって部の 3部を引き出します。肝鎌状間膜、肝周囲の結合織を解剖します。分離し、6-0 絹縫合糸と左下横隔静脈を結紮します。</li>
	<li>胆管にステントを挿入します。<ol>
<li>胃十二指腸静脈が門脈を結合点のすぐ上の BD をクランプし、microforceps を使用して、少しの緊張を維持するために引っ張ります。マイクロはさみで約 5 mm 胆道合流点に近位の前の壁に 1 ミリ長さの"V"切開を行います。</li>
		<li>湾曲した microforceps を用いた胆管の内腔にステントを挿入します。胆管外ステント嘘の少なくとも半分を確認します。</li>
		<li>6-0 シルク縫合糸でステントを保護します。縫合糸の一方の端をカットします。後吻合中に保持するために他の 1 つを保ちます。トランセクト ステント下胆管、胆管ステントから来ることができるかどうかを確認します。</li>
	</ol>
</li>
	<li>左腎静脈レベルまでの IHVC を分離します。IHVC の白骨まで綿芽解離により前方と外側の周囲組織から IHVC を解放します。分離し、右副腎静脈と 6-0 シルク縫合糸で右腎静脈を結紮します。</li>
	<li>陰茎背静脈を通して生理食塩水 2 mL で希釈したヘパリンの注入 50 U でラットを heparinize します。</li>
	<li>肝灌流します。<ol>
<li>左腎静脈下腹部大動脈を分離します。22 G のカテーテルを大動脈に挿入します。</li>
		<li>開胸術を実行し、microforceps を使用して胸部下行大動脈のクランプします。</li>
		<li>3 mL/分の速度で輸液ポンプとヘパリン (4 ° C) の 25 U を含む生理食塩水 20 mL で大動脈にカテーテルを通してドナーの肝臓を灌流します。一方で、肝臓の外に流す灌流液を許可するように右腎静脈下大静脈をカットします。</li>
		<li>ドナーの肝臓になった青白いとき横隔膜の一部とともにグラフトと左腎静脈レベルで IHVC を縦断面します。</li>
	</ol>
</li>
	<li>2 つのマイクロ鉗子で PV を囲む結合組織を解剖します。縛るし、8-0 プロレン縫合糸で幽門静脈を分割します。脾静脈のレベルで太陽光発電を縦断面します。ドナーの肝臓を切除し、冷たい生理食塩水ですすいでください。冷たい生理食塩水では 4 ° C をお風呂の場所</li>
</ol>3. 移植準備
注: 肝移植の準備のためのすべての手順は、4 ° C で冷たい生理食塩水のお風呂で実行
<ol><li>カフをドナーの IHVC (図 3) に接続します。<ol>
<li>そのカフ肝臓に向かって指してカフ拡張子を持つ、IHVC に挿入します。IHVC の後部の壁にカフの拡張機能を配置します。お風呂コンテナー ファイルのクリップを使用して接続されている自体曲線 microforceps でカフの拡張と、IHVC を固定します。</li>
		<li>2 つのマイクロ鉗子によるカフの体の上、IHVC の遠位端を片脚します。</li>
		<li>6-0 絹縫合糸の端を固定します。処理中に、IHVC がねじれていないことを確認します。</li>
	</ol>
</li>
	<li>ドナーの PV にカフを付ける PV の 3.1 の手順を繰り返します。</li>
	<li>磁気リングをドナーのグラフト (図 4) に接続します。<ol>
<li>4 つの鉗子を介してダイヤフラムを引っ張るし、グラフトをトリムします。</li>
		<li>チタン合金マイクロ鉗子を用いた磁気リングを通してグラフトを渡します。</li>
		<li>グラフトの遠位端を輪をかけて片脚します。</li>
		<li>6-0 絹縫合糸が付いているリングを固定します。グラフトは、プロセス中にねじれていないことを確認します。</li>
		<li>肝臓の周り余分なダイヤフラムを削除します。</li>
	</ol>
</li>
	<li>冷たい通常の生理食塩水のお風呂でドナーの肝臓を浸すし、4 ° C で保存</li>
</ol>4. 受信者操作
注: 受信者のラット移植注入のスキーマは図 5に示すです。
<ol><li>注入ブプレノルフィン (0.05 mg/kg 皮下鎮痛剤として操作の前に 1 時間。イソフルラン吸入によるドナー ラットを麻酔します。5 vol % イソフルラン麻酔および流量 0.6 0.8 で 2 vol % イソフルランの誘導のプレキシ グラス箱に 1.5 L/分の流量で L/円錐分マスク麻酔の維持のため。</li>
	<li>ラットの腹部の肌全体を剃る。ポビドン ヨード液と対応する皮膚を消毒します。</li>
	<li>4 cm 正中切開を介して腹腔内を開きます。腹部のリトラクターを配置します。正中線からできるだけ離れて肋骨を引き出します。</li>
	<li>クランプ、止血鉗子を用いた頭に向かって部の 3部を引き出します。肝臓の周りの靭帯、結合組織を解剖します。縛るし、肝食道靱帯を分割します。縛るし、8-0 プロレン縫合糸で肝動脈を分割します。</li>
	<li>最初の肝内門脈から肝内胆管を分離します。その部門の下の総胆管周囲 6-0 絹糸を置き、総胆管を結紮します。</li>
	<li>右腎静脈に IHVC を解剖します。縛るし、右副腎静脈を分割します。肝グラフトにストラップを配置します。</li>
	<li>IHVC 微小血管クリップで右腎静脈の真上をクランプします。微小血管クリップ幽門静脈レベルで太陽光発電をクランプします。<ol>
<li>肝臓から血液をフラッシュ、ゆっくり PV を通して生理食塩水入りの 2 mL を注入します。以前に配置したストラップを引いて肝臓をドラッグします。Satinsky クランプ付きグラフトと横隔膜の部分をクランプします。</li>
		<li>実質の近くに IHVC と肝門部の PV を縦断面します。Transecting グラフト肝のすぐ上で、肝臓を切除します。受信者の肝臓をすぐに削除します。</li>
	</ol>
</li>
	<li>受信者のグラフトに磁気リングを取り付けます。<ol>
<li>磁気リングに受信者の残りのグラフトを挿入します。</li>
		<li>Satinsky クランプの磁気吸引力によって磁気リングを修正します。</li>
		<li>容器は、リングの端をカバーするまで、磁気リングをグラフトを片脚します。</li>
	</ol>
</li>
	<li>ドナー肝移植がんを置き、冷たいぬれたガーゼでカバーします。</li>
	<li>生理食塩水で、ドナーと受信者の SHVCs の内腔を埋めます。空気塞栓を防ぐために徹底的に空気の泡を削除します。グラフト再建を完了するには、磁気力を通して一緒にドナーと受信者のグラフトに埋め込まれた磁性体リングを結合します。</li>
	<li>PV を再構築します。<ol>
<li>PV のテンションを維持するために 8-0 プロレン縫合糸を引いて受信者の PV の左右分岐管。</li>
		<li>幽門静脈から静脈を完全に公開するための脾静脈にダウン受信者の PV のクランプを移動します。</li>
		<li>受信者の前壁を切開します。PV の内腔を生理食塩水で満たしなさい。</li>
		<li>カーブタイプ鉗子とドナーの PV のカフの拡張子を保持します。</li>
		<li>受信者の生食による太陽光発電ルーメンのフラッシュを維持しながら PV にドナーの PV のカフ本体を挿入します。</li>
		<li>6-0 絹糸を結紮、円周で固定します。太陽光発電とグラフトのクランプを外して移植肝臓への血流を再確立します。</li>
	</ol>
</li>
	<li>IHVC、IHVC を再構築するために 4.11 手順を繰り返します。</li>
	<li>ステント法による胆管を再建します。<ol>
<li>受信者の胆管の前方の壁に"V"を切開します。</li>
		<li>受信者 BD の内腔にドナー ステントを挿入し、6-0 周絹糸でそれを修正します。2 胆管を一緒に縫合糸を引き、お互いに近い BD に縫合糸を結ぶ。</li>
		<li>大網組織を胆汁の漏れを防ぐために吻合に折り返します。</li>
	</ol>
</li>
	<li>温かい生理食塩水を腹腔内に水を引きます。3-0 非吸収性の外科的縫合連続の二層で腹部切開を閉じる。</li>
	<li>すぐに 16 mg/kg セフロキシム (抗生物質)、0.05 mg/kg ブプレノルフィン (鎮痛剤) を皮下注射、術後後管理します。</li>
	<li>きれいなケージでラットを配置し、食品を許可する自由水します。</li>
	<li>注入セフロキシム (16 mg/kg)、ブプレノルフィン (0.05 mg/kg 腹腔内 3 日間 12 時間ごと。</li>
</ol>

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肝移植で多くの臨床進行は動物研究に起因することができます。ラット OLT は、臓器保存、移植免疫学、生理学、病理学の研究に広く使われている、よく受け入れられたモデルです。しかし、それは非常に複雑なプロシージャではまた、顕微鏡の高度なスキルが必要です。ラット OLT の手順で多くの改善にもかかわらずグラフト吻合困難なステップに残った。この記事でラット OLT グラフト再?...

同所性肝移植 (OLT) のモデルラットは移植研究1,2のために不可欠です。OLT の最初のラットは、李ら1973年3によって記述されていた。このモデルで全ての船舶は手縫合技術が再建されました。手縫合技術は高度な顕微鏡技術、その利用を大幅に制限する必要があります。それ以来、ラット OLT のプロトコルを元に様々 な変更が報告されています。再建術4 大幅簡体字とは、その中でも、カフ門脈 (太陽光発電) ・鎌田らによって 1979 年に報告された infrahepatic 上大静脈 (IHVC) 吻合法はこのモデルから大幅に改善とみなされます.ただし、受信者の suprahepatic 上大静脈 (グラフト) の短いの前方壁のためカフ テクニックがグラフト再建に使用する非常に困難です。この分野のほとんどの研究者はまだラット OLT5,6,7でボトルネック ステップになりますグラフト再建手縫合手法を使用します。
磁気吻合法 (すなわちmagnamosis) は、2 隻または 2 つ磁石8,9,10,11の間の吸引力を使用して他の管状構造物への接続方法です。磁気力は徐々 に圧縮し、強力なステリ吻合12,13に組織を改造します。この圧縮吻合は、人間14,15で効果的な実証されています。我々 はラットのグラフト吻合用磁気リングのペアを設計しています。私たちの最近の研究は、磁気吻合技術がラット OLT16グラフト再建手縫合法より優れていることを示しています。この記事の目的は、新規磁気吻合法を用いたラットにおけるグラフト再建のため詳細なステップバイ ステップのプロトコルを提供することです。

<table><tbody><tr><td>Anesthesia Machine</td><td>Harvard</td><td>&amp;nbsp;tabletop</td><td>Animal anaesthesia</td></tr><tr><td>PLX7000B HF Mobile Digital C-arm System</td><td>Perlong Medical</td><td>PLX7000B</td><td>It is mainly used for the angiography and photography of various operations</td></tr><tr><td>Syringe Pump</td><td>Mindray</td><td>BeneFusion SP5</td><td>intravenous infusion</td></tr><tr><td>Isoflurane</td><td>RWD life Science Co.</td><td></td><td>anesthetic:for the induction and maintenanceof anesthesia</td></tr><tr><td>iohexol</td><td>Shanghai General Pharmaceutical Co</td><td></td><td>&amp;nbsp;intravascular contrast media&amp;nbsp;&amp;nbsp;</td></tr><tr><td>heparin sodium injection</td><td>SPH No.1 Biochemical &amp;amp; Pharmaceutical Co., LTD&amp;nbsp;</td><td></td><td>&amp;nbsp;prevent the formation of thrombosis&amp;nbsp;</td></tr><tr><td>cefuroxime&amp;nbsp;</td><td>Glaxo Operations UK Limited&amp;nbsp;</td><td></td><td>an antibiotic</td></tr><tr><td>buprenorphine&amp;nbsp;</td><td>TIPR Pharmaceutical Responsible Co.,Ltd</td><td></td><td>an analgesic</td></tr><tr><td>curved microforceps</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>W40350</td><td>surgical tool</td></tr><tr><td>hemostatic forceps(straight)</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>J31010</td><td>surgical tool</td></tr><tr><td>hemostatic forceps(curved)</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>J31020</td><td>surgical tool</td></tr><tr><td>Satinsky clamp</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>XEC050</td><td>surgical tool</td></tr><tr><td>needle holder</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>J32010</td><td>surgical tool</td></tr><tr><td>microneedle holder</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>WBA040</td><td>surgical tool</td></tr><tr><td>notched forceps</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>J42010</td><td>surgical tool</td></tr><tr><td>tissue forceps(with hook)</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>J41010</td><td>surgical tool</td></tr><tr><td>tissue scissor</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>Y00040</td><td>surgical tool</td></tr><tr><td>surgical scissors</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>Y00030</td><td>surgical tool</td></tr><tr><td>micro scissors</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>MR-S121T</td><td>surgical tool</td></tr><tr><td>microvessel clips</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>XEC240</td><td>surgical tool</td></tr><tr><td>straight microforceps(titanium alloy)</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>WCC010</td><td>surgical tool</td></tr><tr><td>curved microforceps&amp;nbsp; (titanium alloy)</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>WCC020</td><td>surgical tool</td></tr></tbody></table>

a rat model of orthotopic liver transplantation using a novel magnetic anastomosis technique for suprahepatic vena cava reconstruction

同所性肝移植 (OLT) のモデルラットは移植研究のために不可欠です。非常に洗練された動物モデルで、急な学習曲線が必要です。門脈 (PV) と infrahepatic 上大静脈 (IHVC) の吻合のためカフ手法の導入は、ラットの移植手順を大幅に簡略化は。ただし、受信者の suprahepatic 上大静脈 (グラフト) の短いの前方壁のためカフ テクニックがグラフト再建に使用する非常に困難です。この分野のほとんどの研究者はまだラット同所性肝移植におけるボトルネックのステップはそれにグラフト再建手縫合手法を使用します。磁気吻合法 (すなわちmagnamosis) は、2 つの磁石との吸引力を使用して 2 つの容器を接続する方法です。私たちの最近の研究は、磁気吻合技術がラットにおけるグラフト再建手縫合法より優れていることを示しています。この記事で新規磁気吻合法を用いたラットにおけるグラフト再建の手順を追ってプロトコルを示す.このモデルでステント法による胆管 (BD) の再建を行った間に、太陽光発電と IHVC の再建が標準カフによる実行されました。肝再動脈化術は実行されませんでした。磁気吻合技術に簡単にグラフト再建を行った、anphepatic 相を大幅短縮します。合理的な学習曲線の後、高度な顕微鏡技術なしも研究者は OLT のこのモデルラットを使用して信頼性が高く、再現性のある結果を生成できます。

約 10 回試行後グラフト再建のため磁気吻合法正常に事前のトレーニングがなかった者によって習得されました。グラフト再建は、2 分未満を取った。受信者のラットの無肝期は約 10 分です。血栓症、出血または angiostegnosis られなかったグラフト吻合部に 1, 5, 30 日再灌流後に。大静脈栓塞は、移植後 2 週間で行われました。図 6のように、磁性?...

Watch this Scientific Journal Video about A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction at JoVE.com

A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction

Suprahepatic 下大静脈 (グラフト) の再建のままラット同所性肝移植における困難なステップです。この記事で新規磁気吻合法を用いたラットにおけるグラフト再建の手順を追ってプロトコルを示す.

同所性同種のモデルラット Suprahepatic 上大静脈再建のため新規磁気吻合法を用いた肝移植

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep ...

a-rat-model-orthotopic-liver-transplantation-using-novel-magnetic

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JoVE Journal

Medicine

9.8K ビュー.  First Affiliated Hospital, Xi'an Jiaotong University. Suprahepatic 下大静脈 (グラフト) の再建のままラット同所性肝移植における困難なステップです。この記事で新規磁気吻合法を用いたラットにおけるグラフト再建の手順を追ってプロトコルを示す.

ビデオ: A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction

Orthotopic Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat liver transplantation in 1979 by Kamada et al.3, this model has remained the gold standard despite various proposed alternative techniques4. Nevertheless, its broader use is limited by its steep learning curve5.
In this video paper, we show a simple and easy-to-establish revision of Kamada's two-cuff technique. The suprahepatic vena cava anastomosis is performed manually with a running suture, and the vena porta and infrahepatic vena cava anastomoses are performed utilizing a quick-linker cuff system6. Manufacturing the quick-linker kit is shown in a separate video paper.

We present an easy-to-establish revision of the classical two-cuff technique for orthotopic liver transplantation in rat.

ラット同所性肝移植

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

医学

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

肝動脈再建を用いた部分同所性肝移植のラットモデルのための外科的処置

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the failing native liver has regenerated.1-3 The APOLT-method, the orthotopic implantation of auxiliary segments- averts most of the technical problems. However this method necessitates extensive resections of both the native liver and the graft.4 In 1998, Erhard developed the heterotopic auxiliary liver transplantation (HALT) utilizing portal vein arterialization (PVA) (Figure 1). This technique showed promising initial clinical results.5-6 We developed a HALT-technique with flow-regulated PVA in the rat to examine the influence of flow-regulated PVA on graft morphology and function (Figure 2).
A liver graft reduced to 30 % of its original size, was heterotopically implanted in the right renal region of the recipient after explantation of the right kidney.&#160; The infra-hepatic caval vein of the graft was anastomosed with the infrahepatic caval vein of the recipient. The arterialization of the donor&#8217;s portal vein was carried out via the recipient&#8217;s right renal artery with the stent technique. The blood-flow regulation of the arterialized portal vein was achieved with the use of a stent with an internal diameter of 0.3 mm. The celiac trunk of the graft was end-to-side anastomosed with the recipient&#8217;s aorta and the bile duct was implanted into the duodenum. A subtotal resection of the native liver was performed to induce acute hepatic failure. 7
In this manner 112 transplantations were performed. The perioperative survival rate was 90% and the 6-week survival rate was 80%. Six weeks after operation, the native liver regenerated, showing an increase in weight from 2.3&#177;0.8 g to 9.8&#177;1 g. At this time, the graft&#8217;s weight decreased from 3.3&#177;0.8 g to 2.3&#177;0.8 g.
We were able to obtain promising long-term results in terms of graft morphology and function. HALT with flow-regulated PVA reliably bridges acute hepatic failure until the native liver regenerates.

Auxiliary liver transplantation provides a temporary support in acute hepatic failure, until regeneration of the failing liver. The heterotopic auxiliary liver transplantation (HALT) with portal vein arterialization (PVA) renders sufficient liver function. We developed an analogous technique in the rat, to examine the influence of the portal vein arterialization on the morphology and function of the graft.

急性肝障害における流れ調節性門脈動脈血と異補助ラット肝移植

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the ...

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 ...

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. To better understand the mechanism behind these surgical procedures, a rat partial liver transplantation (PLTx) model was established for relevant surgical studies. Because of the complexity of the rat PLTx model, a protocol with detailed descriptions is required. An article published previously reported a protocol in which ex vivo hepatectomy was used to achieve 50% rat PLTx. In contrast to this protocol, we introduced a re-arterialized PLTx with an in vivo 70% hepatectomy. An updated vessel-oriented hepatectomy was incorporated into the rat PLTx to refine the microsurgical procedure. The portal veins and hepatic arteries of the left lateral lobe and the median lobe were individually dissected and ligated before removal of the liver parenchyma, thereby decreasing the probability of bleeding in the remnant liver stump. Furthermore, an end-to-side vessel anastomosis between the common hepatic artery and the enlarged proper hepatic artery was introduced to re-arterialize the hepatic artery. By using this end-to-side vessel anastomosis technique, the diameter of the anastomosis was enlarged, thereby decreasing the difficulty of hand suture and maintaining a high rate of anastomotic patency. Moreover, the cuff anastomosis of the infrahepatic vena cava was slightly modified. A section of circumferential liver parenchyma around the vena cava of a recipient was preserved during cuff anastomosis to maintain the three-dimensional shape of the vascular lumen. This section of liver parenchyma was removed after completing the anastomosis. With this modification, the step involving placement of stay sutures was omitted, thereby further shortening the cuff anastomosis time. By using this protocol of rat PLTx, a low liver enzyme level, an intact liver lobular architecture and a high survival rate were achieved after microsurgery.

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Here, a protocol involving re-arterialized rat partial liver transplantation is presented. Specifically, 70% liver was resected in vivo by using an updated technique of vessel-oriented hepatectomy. The hepatic artery was reconstructed in an end-to-side manner. The cuff technique was modified to shorten the anastomosis time of the infrahepatic vena cava.

再門脈部分肝移植のラット体内血管指向 70% 肝切除

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. ...

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation of human liver transplantation due to the absence of arterial reconnection. Described here is a modified transplantation procedure that includes the incorporation of hepatic artery (HA) reconnection, leading to a marked improvement in transplant outcomes. With a mean anhepatic time of 12 min and 14 s, HA reconnection results in improved perfusion of the transplanted liver and an increase in long-term recipient survival from 37.5% to 88.2%. This protocol includes the use of 3D-printed cuffs and holders to connect the portal vein and infrahepatic inferior vena cava. It can be implemented for studying multiple aspects of liver transplantation, from immune response and infection to technical aspects of the procedure. By incorporating a simple and practical method for arterial reconnection using a microvascular technique, this modified rat OLT protocol closely mimics aspects of human liver transplantation and will serve as a valuable and clinically relevant research model.

The goal of this study is to modify the rat orthotopic liver transplant model to better represent human liver transplantation and improve recipient survival. The presented method reestablishes hepatic arterial inflow by connecting the donor liver&#39;s common hepatic artery to the recipient liver&#39;s proper hepatic artery.

ラット型異形肝移植モデルにおける動脈リコネクション後の合併症の減少

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...

Organ Ischemia-Reperfusion Injury by Simulating Hemodynamic Changes in Rat Liver Transplant Model

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, including ischemia-reperfusion injury (IRI) of extrahepatic organs. This model requires numerous experiments and devices. The duration of anhepatic phase is closely related to the time to develop IRI after transplantation. In this experiment, we used hemodynamic changes to induce extrahepatic organ damage in rats and determined the maximum tolerance time. The time until the most severe organ injury varied for different organs. This method can easily be replicated and can also be used to study IRI of the extrahepatic organs after liver transplantation.

This paper provides a detailed description of how to build an animal model of the anhepatic phase (liver ischemia) in rats to facilitate basic research into ischemia-reperfusion injury after liver transplantation.

ラット肝移植モデルにおける血球力学的変化をシミュレートすることによる臓器虚血再灌流損傷

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, ...

Porcine Liver Transplantation Without Veno-Venous Bypass As an Extended Criteria Donor Model

Liver transplantation is regarded as the gold standard for the treatment of a variety of fatal hepatic diseases. However, unsolved issues of chronic graft failure, ongoing organ donor shortages, and the increased use of marginal grafts call for the improvement of current concepts, such as the implementation of organ machine perfusion. In order to evaluate new methods of graft reconditioning and modulation, translational models are required. With respect to anatomical and physiological similarities to humans and recent progress in the field of xenotransplantation, pigs have become the main large animal species used in transplantation models. After the initial introduction of a porcine orthotopic liver transplant model by Garnier et al. in 1965, several modifications have been published over the past 60 years.
Due to specifies-specific anatomical traits, a veno-venous bypass during the anhepatic phase is regarded as a necessity to reduce intestinal congestion and ischemia resulting in hemodynamic instability and perioperative mortality. However, the implementation of a bypass increases the technical and logistical complexity of the procedure. Furthermore, associated complications such as air embolism, hemorrhage, and the need for a simultaneous splenectomy have been reported previously.
In this protocol, we describe a model of porcine orthotopic liver transplantation without the use of a veno-venous bypass. The engraftment of donor livers after static cold storage of 20 h - simulating extended criteria donor conditions - demonstrates that this simplified approach can be performed without significant hemodynamic alterations or intraoperative mortality and with regular uptake of liver function (as defined by bile production and liver-specific CYP1A2 metabolism). The success of this approach is ensured by an optimized surgical technique and a sophisticated anesthesiologic volume and vasopressor management.
This model should be of special interest for workgroups focusing on the immediate postoperative course, ischemia-reperfusion injury, associated immunological mechanisms, and the reconditioning of extended criteria donor organs.

In this protocol, a model of porcine orthotopic liver transplantation after static cold storage of donor organs for 20 h without the use of a veno-venous bypass during engraftment is described. The approach uses a simplified surgical technique with minimization of the anhepatic phase and sophisticated volume and vasopressor management.

拡張基準ドナーモデルとしての静脈バイパスなしのブタ肝移植

Liver transplantation is regarded as the gold standard for the treatment of a variety of fatal hepatic diseases. However, unsolved issues of chronic graft ...

マウスへの肝移植と同種移植片の開存性の分析

Successful Orthotopic Liver Transplantation in Mice Utilizing Microcomputed Tomography Angiography

Microcomputed tomography (microCT) angiography is an invaluable resource to researchers. New advances in this technology have allowed for high-quality images to be obtained of micro-vasculature and are high-fidelity tools in the field of organ transplantation. In this model of orthotopic liver transplantation (OLT) in mice, microCT affords the opportunity to evaluate allograft anastomosis in real time and has the added benefit of not having to sacrifice study animals. The choice of contrast, as well as image acquisition settings, create a high-definition image, which gives researchers invaluable information. This allows for evaluation of the technical aspects of the procedure as well as potentially evaluating different therapeutics over an extended duration of time. In this protocol, we detail an OLT model in mice in a stepwise fashion and finally describe a microCT protocol that can give high-quality images, which aid researchers in in-depth analysis of solid organ transplantation. We provide a step-by-step guide for liver transplantation in a mouse, as well as briefly discuss a protocol for evaluating the patency of the graft through microCT angiography.

著者スポットライト:マウスモデルにおけるMicroCT血管造影による移植研究の強化 

In this protocol, we discuss the implementation of a model of successful orthotopic liver transplantation (OLT) in mice. Additionally, adjuvants to further analyze allograft patency after successful OLT in a mouse are discussed as well, specifically utilizing microcomputed tomography (microCT) scans.

マイクロコンピュータ断層撮影血管造影を用いたマウスの同所性肝移植の成功

Microcomputed tomography (microCT) angiography is an invaluable resource to researchers. New advances in this technology have allowed for high-quality ...

同所性同種のモデルラット Suprahepatic 上大静脈再建のため新規磁気吻合法を用いた肝移植

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ラット同所性肝移植

肝動脈再建を用いた部分同所性肝移植のラットモデルのための外科的処置

急性肝障害における流れ調節性門脈動脈血と異補助ラット肝移植

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

再門脈部分肝移植のラット体内血管指向 70% 肝切除

ラット型異形肝移植モデルにおける動脈リコネクション後の合併症の減少

ラット肝移植モデルにおける血球力学的変化をシミュレートすることによる臓器虚血再灌流損傷

拡張基準ドナーモデルとしての静脈バイパスなしのブタ肝移植

マイクロコンピュータ断層撮影血管造影を用いたマウスの同所性肝移植の成功

マイクロコンピュータ断層撮影血管造影を用いたマウスの同所性肝移植の成功