この研究は、「ドイツ学術振興」（DFG）によって資金を供給された。 K. SchleimerはNRW州のリセマイトナー科学省のフェローシップや研究によってサポートされていました。私たちは彼女の優れた編集支援のため夫人メアリー·ジョアンBlümichに感謝したいと思います。博士マリアKokozidouはフレゼニウス2011_A61から融資された。

1。実験動物<ol><li>動物保護FELASAガイドライン（K 40、99分の17）に応じて、実行されるすべての実験のための地域倫理委員会の承認を取得します。 </li><li>商業ブリーダー（ 例えば、チャールズリバーWiga社、ズルツフェルト、ドイツ）から購入可能オスのルイスラットを使用してください。ここでは、224の動物（112ドナー、レシピエント112）は112の移植を行うために使用された。受信者は術後6週間で屠殺した。動作時には、動物は250〜350グラムの重量を量る必要があります。 </li><li>食物と水に自由にアクセスさせて気候制御室で動物を保​​管してください。 </li></ol> 2麻酔と手術の準備<ol><li> 1リットル/分の酸素流量、および麻酔を誘導する5％の麻酔ガスとイソフルラン気化器と麻酔器に接続されたプレキシグラス·ボックスにラットを置きます。 1.5％のイソフルランで麻酔を維持します。 </li><li>ラットを置きます温暖化プレート上仰臥位。 </li><li>脱毛クリームで腹部の毛を削除します。 </li><li>ベンチトップをきれいに消毒剤を使用して、次のように無菌手術野を準備します。最初の切開部位用クロルヘキシジン、アルコール秒と、最終的に、スクラブ溶液（ 例えば、クロルヘキシジン及び70％イソプロピルアルコール）の消毒のため。使い捨て手術着や手術用手袋を着用してください。 </li><li>無菌マイクロサージェリー機器を使用してください。 </li><li> 12-20倍の倍率の手術用顕微鏡を使用してください。 </li></ol> 3。外科的手技<ol><li>ドナー操作 準備 <ol><li>中央値開腹を行い、ヒギンズによって記載されているように左側葉および移植片の中葉を切除。14 </li><li>その後、肝十二指腸間膜を準備します。まず小さな胆管の混雑と拡張を誘導するために、胆管を結紮。その後gastroduodeを連結し、横断するNAL静脈と動脈。その後、腹腔動脈と総肝動脈を分離し、左胃動脈と脾動脈を横断する。混雑した胆管を切開し、その中に20-Gのステントを導入。ステントの位置決めは、全体の肝門部胆管システムの排水を確保する必要があります。ステントに対して遠位胆管を横断する。 </li><li>右右腎静脈の上大静脈の肝臓下のセグメントを分離します。 かん流 </li><li>腹腔動脈上で大動脈のライゲーション後、門脈をクランプし、それを切開。門脈に灌流カテーテル（14 G）を導入し、ヒスチジン - トリプトファン - ケトグルタル酸（HTK） - 溶液（12センチH2O、4℃）30mlで灌流を行う。 </li><li>ダイアフラムと灌流液が流出することを可能にする肝上大静脈をカット。 </li><li>大動脈パッチと一緒に腹腔動脈を切除し、ヒスチジン - トリプトファン - ケトグルタル酸溶液ですすいでください。 植 </li><li>右右腎静脈の上に肝臓下の大静脈を横断する。 </li><li>その後、門脈0.3mmの直径のステントを配置して、合字で固定します。以下の動作中に、門脈は右腎動脈を介して動脈血されています。ステントに遠位に門脈を横断する。 </li><li>肝上大静脈の結紮後、ダイヤフラムのカフと一緒に横断する。 </li><li>移植片を外植し、氷上でビニール袋に4°摂氏ヒスチジン - トリプトファン - ケトグルタル酸（HTK） - 溶液に保管してください。肝臓下の大静脈にコーナー縫合糸（8-0 Ethilon）を配置します。 </li></ol></li><li>受信者の操作<ol><li>中央値開腹を行い、肝臓下の大静脈を準備します。その後、右腎動脈と尿管を隔離尿管を結紮し、それを横断する。その原点で右腎動脈をクランプし、右腎静脈を結紮。その後、両方の離断後に再NALの血管は、腎摘出を行う。続いて、ヘパリン生理食塩水（40 UIヘパリン/ mlの生理食塩水）ですすいで、右腎動脈を短縮する。 </li><li>肝臓下の大静脈に楕円形の内腔を作成するために肝臓下の大静脈の近位および遠位のクランプの後、結紮の近位に、右腎静脈を横断する。ヘパリン生理食塩水で大静脈をすすぐ。受信者の腹部の右腎領域での移植片を移植する。 </li><li> 8-0 Ethilon連続縫合でrecipient`s大静脈に、ドナーの肝臓下の大静脈の端側を吻合。 </li><li>ヘパリン/食塩水ですすいだ後、右腎動脈（ 図3）に門脈ステントをご紹介します。 2ステッチ（8-0 Ethilon）と門脈および腎動脈を接続します。合字（編組シルク黒7-0）で腎動脈内のリンクステントを固定してください。 </li><li>再灌流を可能にするには、まず両方のその後、動脈クランプを削除静脈クランプ（ 図4）。 </li><li>大動脈大動脈を準備し、その後、近位および遠位microclampを配置し、右の両方クランプ間の大動脈を切開。 10-0 Ethilon連続縫合で、大動脈パッチを使用して、大動脈およびドナーの腹腔動脈との間に端側吻合を実行します。 </li><li>十二指腸への胆管を挿入します。十二指腸壁に巾着縫合糸を適用する、右巾着の真ん中に十二指腸壁に小さな切開を行います。十二指腸にステント付胆管を導入し、巾着縫合糸を締めます。最後に、2つのステッチ（Ethilon 8-0）（ 図5）でしっかりと一緒に十二指腸と胆管を開催しています。 </li><li>その後、最初のEmond らによって記載ネイティブ肝臓の小計切除を行う。急性肝不全を模倣する。7 </li><li>再の横方向の腹壁に、一緒にダイアフラムのカフで、移植片の肝上大静脈を修正4-0ビクリル縫合糸でcipient。生理食塩水（ 図6）で腹部を洗い流す。 </li><li>筋層では4-0ビクリル連続縫合し、皮膚のために3-0のVicryl連続縫合を使用しています。 </li></ol></li></ol> 4。回復と術後管理<ol><li>動物は、赤外線加温ランプ下で麻酔から回復しましょう​​。 </li><li> 2日間、8〜12時間の0.03〜0.05 mg / kg体重の皮下の用量でブプレノルフィンを注入する。 </li><li>全身状態や病気や感染症の兆候について動物を調べます。 </li></ol>初期ネイティブ肝重量の5。計算以下のようにして初期ネイティブ肝臓重量を計算し+/- 0.49％をnと以前の実験で決定された標準偏差完全肝臓重量（体重2.66％（この値から切除肝葉の重量を差し引く同じ株であり、a = 80の動物GE））。15

<ol>
	<li>Boudjema, K., 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=Auxiliary+liver+transplantation+for+fulminant+and+subfulminant+hepatic+failure.">Auxiliary liver transplantation for fulminant and subfulminant hepatic failure.</a> Transplantation. 59, 218-223 (1995).</li><li>Boudjema, K., Jaeck, D., Sim&#233;oni, U., Bientz, J., Chenard, M. P., Brunot, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temporary+auxiliary+liver+transplantation+for+subacute+liver+failure+in+a+child.">Temporary auxiliary liver transplantation for subacute liver failure in a child.</a> The Lancet. 342, 778-779 (1993).</li><li>Chenard-Neu, M. P., 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=Auxiliary+liver+transplantation:+Regeneration+of+the+native+liver+and+outcome+in+30+patients+with+fulminant+hepatic+failure+-+a+multicenter+European+study.">Auxiliary liver transplantation: Regeneration of the native liver and outcome in 30 patients with fulminant hepatic failure - a multicenter European study.</a> Hepatology. 23, 1119-1127 (1996).</li><li>Gubernatis, G., Pichlmayr, R., Kemnitz, J., Gratz, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+partial+orthotopic+liver&#8211;transplantation+(APOLT)+for+fulminant+hepatic+failure:+first+successful+case+report.">Auxiliary partial orthotopic liver&#8211;transplantation (APOLT) for fulminant hepatic failure: first successful case report.</a> World J Surg. 15, 660-666 (1991).</li><li>Erhard, J., Lange, R., Giebler, R., Rauen, U., de Groot, H., Eigler, F. 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T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+liver+grafting:+a+new+concept+in+liver+transplantation.">Auxiliary liver grafting: a new concept in liver transplantation.</a> The Lancet. 342, 758 (1993).</li><li>Terpstra, O. T., Reuvers, C. B., Schalm, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+heterotopic+liver+transplantation.">Auxiliary heterotopic liver transplantation.</a> Transplantation. 45, 1003-1007 (1988).</li><li>Schleimer, K., Lange, R., Rauen, U., Erhard, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+liver+transplantation+in+acute+liver+failure+in+the+rat+-+an+illustrated+description+of+a+new+surgical+approach.">Auxiliary liver transplantation in acute liver failure in the rat - an illustrated description of a new surgical approach.</a> Langenbecks Arch Surg. 384 (2), 204-208 (1999).</li><li>Schleimer, K., 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=Physiologic+microcirculation+of+the+heterotopically+transplanted+rat+liver+with+portal+vein+arterialization+depending+on+optimal+stent+diameter.">Physiologic microcirculation of the heterotopically transplanted rat liver with portal vein arterialization depending on optimal stent diameter.</a> Med Sci Monit. 28 (BR140-145), (2006).</li><li>Hong, H. Q., 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=Study+of+portal+arterialization+with+auxiliary+liver+in+rats.">Study of portal arterialization with auxiliary liver in rats.</a> Hiroshima J. Med. Sci. 40, 29-33 (1991).</li><li>Higgins, G. M., Anderson, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+pathology+of+the+liver.+I--Restoration+of+the+liver+of+the+white+rat+following+partial+surgical+removal.">Experimental pathology of the liver. I--Restoration of the liver of the white rat following partial surgical removal.</a> Arch. Path. 12, 186-202 (1931).</li><li>Schleimer, K., 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=Auxiliary+liver+transplantation+with+flow-regulated+portal+vein+arterialization+offers+a+successful+therapeutic+option+in+acute+hepatic+failure--investigations+in+heterotopic+auxiliary+rat+liver+transplantation.">Auxiliary liver transplantation with flow-regulated portal vein arterialization offers a successful therapeutic option in acute hepatic failure--investigations in heterotopic auxiliary rat liver transplantation.</a> Transpl Int. 19 (7), 581-588 (2006).</li><li>Lee, S., Edgington, T. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+transplantation+in+the+rat.">Liver transplantation in the rat.</a> Surg. Forum. 17, 220-222 (1966).</li><li>Hess, F., Willemen, A., Jerusalem, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+liver+transplantation+in+the+rat,+influence+of+the+condition+of+the+recipient's+liver+on+the+fate+of+the+graft.">Auxiliary liver transplantation in the rat, influence of the condition of the recipient's liver on the fate of the graft.</a> Eur. Surg. Res. 9, 270-279 (1977).</li><li>Aguirrezabalaga, J., Arnal, F., Marini, M., Centeno, A., Fernandez-Selles, C., Rey, I., Gomez, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+liver+transplantation+with+portal+arterialization+in+the+rat:+description+of+a+new+model.">Auxiliary liver transplantation with portal arterialization in the rat: description of a new model.</a> Microsurgery. 22, 21-26 (2002).</li><li>Palmes, D., Dietl, K. H., Drews, G., Holzen, J. P., Spiegel, H. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+partial+orthotopic+liver+transplantation:+treatment+of+acute+liver+failure+in+a+new+rat+model.">Auxiliary partial orthotopic liver transplantation: treatment of acute liver failure in a new rat model.</a> Langenbecks Arch Surg. (386), 534-541 (2002).</li><li>Schleimer, K., 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=Improved+microcirculation+of+a+liver+graft+by+controlled+portal+vein+arterialization.">Improved microcirculation of a liver graft by controlled portal vein arterialization.</a> J Surg Res. 116 (2), 202-210 (2004).</li><li>Schleimer, K., 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=Portal+hyperperfusion+causes+disturbance+of+microcirculation+and+increased+rate+of+hepatocellular+apoptosis:+investigations+in+heterotopic+rat+liver+transplantation+with+portal+vein+arterialization.">Portal hyperperfusion causes disturbance of microcirculation and increased rate of hepatocellular apoptosis: investigations in heterotopic rat liver transplantation with portal vein arterialization.</a> Transplant Proc. 38 (3), 725-729 (2006).</li></ol>

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最初の異補助ラット肝臓移植はLee らによって記述されました。、1966年に16はヘスに続く。グラフトの17門脈は、レシピエントの門脈を吻合した。この手順の間、ネイティブの肝臓に門脈血液供給が損なわれます。 1991年には香港は、受信者の腸骨動脈を介してPVAでHALTを行った。13大規模なポータルと正弦波の輻輳が発生した血流量調...

急性肝障害が60〜80％の死亡率につながる臨床医学における深刻な問題となる。急性肝不全で8、補助肝移植は、急性期を埋める有望な代替療法である。ネイティブ肝臓を再生するために残されていると再生に、以下の免疫抑制薬を中止することができます。その結果、移植片は、萎縮性になります。しかし補助肝移植の手術技術はまだ議論されている。1-3この文脈において、テルプストラは移植のためのスペースがないことを報告し、ポータル血液は現象と静脈うっ血や排水の問題を盗む。9-10ドナーの門脈受信者の門脈に、端側を吻合した。これは、移植片とネイティブ肝臓の間の門脈血栓症のポータル流れの競争をもたらした。これらの技術的な問題のほとんどは、APOLT法（orthotopの使用により回避することができる補助セグメントのIC注入）。4しかし、この方法は、ネイティブの肝臓および移植片の両方の広範な切除が必要となる。1998年4、エアハルトと彼のチームは、門脈動脈化（PVA）（ 図1を利用し異補助肝移植（HALT）を開発）。門とネイティブ肝臓の門脈には、この方法を利用して尊重される。すなわち、このような門脈血栓症および門脈血液のような望ましくない副作用から受信者は、ネイティブの肝臓の回復を乱す可能性が現象を、盗むスペア。さらに、ネイティブ肝臓および移植片のない大規模な肝切除は（グラフト）と再生（ネイティブ肝臓）をブリッジするため、結果としてより少ない出血、より残りの総肝実質塊で、この方法のために必要ありません。この技術は、最初の臨床結果を約束しあった。5-6この実験的研究の目標は、門脈動脈化のあるHALT技術の開発だった研究目的のために、ラット（ 図2）。以来11。私たちは考慮に再灌流し、肝臓カプセルから大量出血後に移植片の混雑をもたらした動脈血門脈の血流調節を、かかりませんでした予備実験では動脈血門脈におけるその後の血流0.3mmの内径および8mmの長さのステントによって調節した。パイロット実験は、0.3mmのステント直径は、上部生理学的範囲内にある動脈血門脈における平均血流量をもたらすことを示した（1.7 +/- 0.4ミリリットル/分/ g肝臓重量）。血流量調節せずに12本発明者らの結果受信者の腸骨動脈を介してPVAでHALTを実行した香港の結果と相関している。彼の実験では、13の大規模なポータルおよび正弦波の輻輳が発生した血流量調節なしで。 私達のビデオの目的は、Dにあるステップバイステップemonstrateラットにおけるこのne.w実験的手術法。長期的には提案された技術は、肝再生およびインター肝競争と門脈動脈化、門脈過灌流とポータルの血流量調節の研究で使用することができます。

<table border="0" cellpadding="0" cellspacing="0" width="573">
	<tbody>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Braided Silk black 7-0&#160;</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">892100</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Vicryl 3-0</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">J285G</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Vicryl 4-0</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">J284G</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Ethilon 8-0</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">1714G</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Ethilon 10-0</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">7770G</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Vasofix safety 14G</td>
			<td style="width:71px;">B Braun</td>
			<td style="width:119px;">2758853</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Vasofix safety 20G</td>
			<td style="width:71px;">B Braun</td>
			<td style="width:119px;">2758818</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="201">
			<td height="201" style="height:201px;width:216px;">2 French (.012&#8221;/0.3mm ID x .025&#8221;/0.6mm OD) x 24&#8221;/60cm polyurethane catheter</td>
			<td style="width:71px;">Access technologies a division of Norfolk Medical, Inc., Illinois, USA</td>
			<td style="width:119px;">CNC-2P</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="119">
			<td height="119" style="height:119px;width:216px;">HTK solution,</td>
			<td rowspan="2" style="width:71px;">Dr. Franz K&#246;hler Chemie GmbH, Alsbach-H&#228;hnlein, Germany</td>
			<td rowspan="2" style="width:119px;"></td>
			<td rowspan="2" style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Custodiol</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Isofluorane 5 %</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Any genericon</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Heparin</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Any genericon</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">0.9% saline</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Any genericon</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Buprenorphine</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Any genericon</td>
		</tr>
	</tbody>
</table>

heterotopic auxiliary rat liver transplantation with flow-regulated portal vein arterialization in acute hepatic failure

急性肝不全補助肝移植では興味深い代替アプローチである。目的は、障害のあるネイティブ肝臓が再生するまでの一時的なサポートを提供することです。1-3 APOLT法、補助の区分の同所移植は技術的な問題のほとんどをaverts。しかし、この方法は、ネイティブの肝臓および移植片の両方の広範な切除が必要となる。1998年4、エアハードは門脈動脈化（PVA）（図1）を利用した異補助肝移植（HALT）を開発した。この技術は、最初の臨床結果を約束しあった。5-6私たちは、移植片の形態と機能（図2）上のフローレギュPVAの影響を調べるために、ラットに、フローレギュレートされたPVAとHALT-技術を開発した。 元のサイズの30％に低下し、肝臓移植は、heterotopically右腎の外移植後のレシピエントの右腎領域で移植した。60;移植片の赤外線肝大静脈は、レシピエントの肝臓下の大静脈を吻合した。ドナーの門脈の動脈化は、ステント技術に受信者の右腎動脈を介して行われた。動脈血門脈の血流の調節は、0.3mmの内径を有するステントの使用により達成された。移植片の腹腔動脈は、エンドツー側受信者の大動脈を吻合し、胆管が十二指腸内に注入した。ネイティブ肝臓の小計切除が急性肝不全を誘導するために行った。7 このようにして、112移植を行った。周術期の生存率は90％であり、6週間の生存率は80％であった。六週間運転後に、ネイティブの肝臓は9.8±1グラムに2.3±0.8グラムから重量の増加を示し、再生した。この時点で、移植片の重量が2.3±0.8グラムに3.3±0.8グラムから減少した。 </p&gt; 私たちは、移植片の形態および機能の点で有望な長期的結果を得ることができた。ネイティブ肝臓が再生するまでのフローレギュレートされたPVAとHALT確実急性肝不全を埋める。

移植の平均持続時間は約150分であった。冷虚血時間は、64±7分であった温虚血時間未満、25分であった。 周術期の生存率は90％であり、6週間の生存率は80％であった。 20％の損失には、次の5週間の間、術後1週の間にあるため、ステント脱臼および胆汁の漏れ（4匹）の腹膜炎によるものであったと肺炎、腹腔内膿瘍および腸閉塞（7匹）へ。生存している動物は、優れた一?...

Watch this Scientific Journal Video about Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure at JoVE.com

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

補助肝移植は、失敗した肝臓の再生まで、急性肝不全の一時的なサポートを提供します。門脈動脈化（PVA）と異補助肝移植（HALT）は、十分な肝機能をレンダリングします。私たちは、移植片の形態と機能に門脈動脈化の影響を調べるために、ラットに類似した技術を開発しました。

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 ...

heterotopic-auxiliary-rat-liver-transplantation-with-flow-regulated

Research

JoVE Journal

Medicine

12.6K ビュー.  University Hospital RWTH Aachen. 補助肝移植は、失敗した肝臓の再生まで、急性肝不全の一時的なサポートを提供します。門脈動脈化（PVA）と異補助肝移植（HALT）は、十分な肝機能をレンダリングします。私たちは、移植片の形態と機能に門脈動脈化の影響を調べるために、ラットに類似した技術を開発しました。 

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

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 ...

医学

Rat Heterotopic Abdominal Heart/Single-lung Transplantation in a Volume-loaded Configuration

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft involves anastomosis of donor inferior vena cava (IVC) to recipient IVC, and donor ascending aorta (Ao) to recipient abdominal Ao. The right upper and middle lung lobes are preserved and function as conduits for blood flow from right heart to left heart.
There are several advantages to using this technique, and it lends itself to a broad range of applications. Because the graft is transplanted in a configuration that allows for dyamic volume-loading, cardiac function may be directly assessed in vivo. The use of pressure-volume conductance catheters permits characterization of load-dependent and load-independent hemodynamic parameters. The graft may be converted to a loaded configuration by applying a clamp to the recipient&#8217;s infra-hepatic IVC. We describe modified surgical techniques for both donor and recipient operations, and an ideal myocardial protection strategy. Depending on the experimental aim, this model may be adapted for use in both acute and chronic studies of graft function, immunologic status, and variable ventricular loading conditions. The conducting airways to the transplanted lung are preserved, and allow for acute lung re-ventilation. This facilitates analysis of the effects of the mixed venous and arterial blood providing coronary perfusion to the graft.
A limitation of this model is its technical complexity. There is a significant learning curve for new operators, who should ideally be mentored in the technique. A surgical training background is advantageous for those wishing to apply this model. Despite its complexity, we aim to present the model in a clear and easily applicable format. Because of the physiologic similarity of this model to orthotopic transplantation, and its broad range of study applications, the effort invested in learning the technique is likely to be worthwhile.

We describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The transplant configuration results in a partially loaded graft circulation, allowing direct functional assessment. This model may be employed for acute or chronic studies of function and immunologic status of the transplanted graft.

ラット異腹部ボリュームにロードされた構成でのハート/シングル肺移植

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft ...

Heterotopic Cervical Heart Transplantation in Mice

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies this model by avoiding challenging suture anastomoses of small vessels thereby reducing warm ischemia time. In comparison to abdominal graft implantation the cervical model is less invasive and the implanted graft is easily accessible for further follow-up examinations. Anastomoses are performed by pulling the ascending aorta of the graft over the cuff with the recipient&#8217;s common carotid artery and by pulling the main pulmonary artery over the cuff with the external jugular vein. Selection of appropriate cuff size and complete mobilization of the vessels are important for successful revascularization. Ischemia-reperfusion (I/R) injury can be minimized by perfusing the graft with a cardioplegic solution and by hypothermia. In this article, we provide technical details for a simplified and improved cuff technique, which should allow surgeons with basic microsurgical skills to perform the procedure with a high success rate.

The cuff technique shortens and facilitates the model of heterotopic cervical heart transplantation in mice by avoiding technically challenging suture anastomoses of small vessels. The technical details shown in this video paper should allow researches to establish this model in their laboratories.

マウスにおける異子宮頸心臓移植

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies ...

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) Procedure

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver metastases, the amount of liver tissue left behind after liver resection has become the main limiting factor of resectability of large or multiple liver tumors. A minimal amount of functional tissue is required to avoid the severe complication of post-hepatectomy liver failure, which has high morbidity and mortality. Inducing liver growth of the prospective remnant prior to resection has become more established in liver surgery, either in the form of portal vein embolization by interventional radiologists or in the form of portal vein ligation several weeks prior to resection. Recently, it was shown that liver regeneration is more extensive and rapid, when the parenchymal transection is added to portal vein ligation in a first stage and then, after only one week of waiting, resection performed in a second stage (Associating Liver Partition and Portal vein ligation for Staged hepatectomy = ALPPS). ALPPS has rapidly become popular across the world, but has been criticized for its high perioperative mortality. The mechanism of accelerated and extensive growth induced by this procedure has not been well understood. Animal models have been developed to explore both the physiological and molecular mechanisms of accelerated liver regeneration in ALPPS. This protocol presents a rat model that allows mechanistic exploration of accelerated regeneration.

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy ALPPS Procedure

Inducing rapid liver hypertrophy using Associating Liver Partition and Portal vein ligation for a Staged hepatectomy (ALPPS) has been proposed for resection of borderline resectable liver tumors. This model may elucidate mechanisms involved in rapid hypertrophy and allows testing of drugs that promote or block the acceleration of regeneration.

肝のパーティションを関連付けると段階的な肝切除 (ALPPS) プロシージャの門脈結紮ラットモデル

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver ...

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. ...

Invasive Hemodynamic Characterization of the Portal-hypertensive Syndrome in Cirrhotic Rats

This is a detailed protocol describing invasive hemodynamic measurements in cirrhotic rats for the characterization of portal hypertensive syndrome. Portal hypertension (PHT) due to cirrhosis is responsible for the most severe complications in patients with liver disease. The full picture of the portal hypertensive syndrome is characterized by increased portal pressure (PP) due to the increased intrahepatic vascular resistance (IHVR), hyperdynamic circulation, and increased splanchnic blood flow. Progressive splanchnic arterial vasodilation and increased cardiac output with elevated heart rate (HR) but low arterial pressure characterizes the portal hypertensive syndrome.
Novel therapies are currently being developed that aim to decrease PP by either targeting IHVR or increased splanchnic blood flow &#8212; but side effects on systemic hemodynamics may occur. Thus, a detailed characterization of portal venous, splanchnic, and systemic hemodynamic parameters, including measurement of PP, portal venous blood flow (PVBF), mesenteric arterial blood flow, mean arterial pressure (MAP), and HR is needed for preclinical evaluation of the efficacy of novel treatments for PHT. Our video article provides the reader with a structured protocol for performing invasive hemodynamic measurements in cirrhotic rats. In particular, we describe the catheterization of the femoral artery and the portal vein via an ileocolic vein and the measurement of portal venous and splanchnic blood flow via perivascular Doppler-ultrasound flow probes. Representative results of different rat models of PHT are shown.

Here we describe a detailed protocol for invasive measurements of hemodynamic parameters including portal pressure, splanchnic blood flow, and systemic hemodynamics in order to characterize the portal hypertensive syndrome in rats.

肝硬変ラットにおけるポータル高血圧症侵襲血行動態評価

This is a detailed protocol describing invasive hemodynamic measurements in cirrhotic rats for the characterization of portal hypertensive syndrome. ...

Generation of a Rat Model of Acute Liver Failure by Combining 70% Partial Hepatectomy and Acetaminophen

Acute liver failure (ALF) is a clinical condition caused by various etiologies resulting in the loss of metabolic, biochemical, synthesizing, and detoxifying functions of the liver. In most irreversible liver damage cases, orthotropic liver transplant (OLT) remains the only available treatment. To study the therapeutic potential of a treatment for ALF, its prior testing in an animal model of ALF is essential. In the current study, an ALF model in rats was developed by combining 70% partial hepatectomy (PHx) and injections of acetaminophen (APAP) that provides a therapeutic window of 48 h. The median and left lateral lobes of the liver were removed to excise 70% of the liver mass and APAP was given 24 h postsurgically for 2 days. Survival in ALF-induced animals was found to be severely decreased. The development of ALF was confirmed by altered serum levels of the enzymes alanine amino transferase (ALT), aspartate amino transferase (AST), alkaline phosphatase (ALP); changes in prothrombin time (PT); and assessment of the international normalized ratio (INR). Study of the gene expression profile by qPCR revealed an increase in expression levels of genes involved in apoptosis, inflammation, and in the progression of liver injury. Diffused degeneration of hepatocytes and infiltration of immune cells was observed by histological evaluation. The reversibility of ALF was confirmed by the restoration of survival and serum levels of ALT, AST, and ALP after intrasplenic transplantation of syngeneic healthy rat hepatocytes. This model presents a reliable alternative to the available ALF animal models to study the pathophysiology of ALF as well as to evaluate the potential of a novel therapy for ALF. The use of two different approaches also makes it possible to study the combined effect of physical and drug-induced liver injury. The reproducibility and feasibility of current procedure is an added benefit of the model.

The acute liver failure animal model developed in the current study presents a feasible alternative for the study of potential therapies. The current model employs the combined effect of physical and drug-induced hepatic injury and provides a suitable time window to study the potential of novel therapies.

70%部分肝切除術とアセトアミノフェンを組み合わせた急性肝不全ラットモデルの生成

Acute liver failure (ALF) is a clinical condition caused by various etiologies resulting in the loss of metabolic, biochemical, synthesizing, and ...

Minimizing Post-Infusion Portal Vein Bleeding during Intrahepatic Islet Transplantation in Mice

Although the liver is currently accepted as the primary transplantation site for human islets in clinical settings, islets are transplanted under the kidney capsule in most rodent preclinical islet transplantation studies. This model is commonly used because murine intrahepatic islet transplantation is technically challenging, and a high percentage of mice could die from surgical complications, especially bleeding from the injection site post-transplantation. In this study, two procedures that can minimize the incidence of post-infusion portal vein bleeding are demonstrated. The first method applies an absorbable hemostatic gelatin sponge to the injection site, and the second method involves penetrating the islet injection needle through the fat tissue first and then into the portal vein by using the fat tissue as a physical barrier to stop bleeding. Both methods could effectively prevent bleeding-induced mouse death. The whole liver section showing islet distribution and evidence of islet thrombosis post-transplantation, a typical feature for intrahepatic islet transplantation, were presented. These improved protocols refine the intrahepatic islet transplantation procedures and may help laboratories set up the procedure to study islet survival and function in pre-clinical settings.

Here we present refined surgical procedures on successfully performing intraportal islet transplantation, a clinically relevant but technically challenging surgical procedure, in mice.

マウスの肝内膵島移植中の注入後門脈静脈出血の最小化

Although the liver is currently accepted as the primary transplantation site for human islets in clinical settings, islets are transplanted under the ...

Rat Model of Normothermic Ex-Situ Perfused Heterotopic Heart Transplantation

Heart transplantation is the most effective therapy for end-stage heart failure. Despite the improvements in therapeutic approaches and interventions, the number of heart failure patients waiting for transplantation is still increasing. The normothermic ex situ preservation technique has been established as a comparable method to the conventional static cold storage technique. The main advantage of this technique is that donor hearts can be preserved for up to 12 h in a physiologic condition. Moreover, this technique allows resuscitation of the donor hearts after circulatory death and applies required pharmacologic interventions to improve donor function after implantation. Numerous animal models have been established to improve normothermic ex situ preservation techniques and eliminate preservation-related complications. Although large animal models are easy to handle compared to small animal models, it is costly and challenging. We present a rat model of normothermic ex situ donor heart preservation followed by heterotopic abdominal transplantation. This model is relatively cheap and can be accomplished by a single experimenter.

Here, we present an assessment protocol of a heterotopically implanted heart after normothermic ex situ preservation in the rat model.

ノルマザーミックEx-situ灌流異所性心臓移植のラットモデル

Heart transplantation is the most effective therapy for end-stage heart failure. Despite the improvements in therapeutic approaches and interventions, the ...

内皮から間葉への移行を研究するための生体内心臓移植

A Neonatal Heterotopic Rat Heart Transplantation Model for the Study of Endothelial-to-Mesenchymal Transition

Endocardial fibroelastosis (EFE), defined by subendocardial tissue accumulation, has major impacts on the development of the left ventricle (LV) and precludes patients with congenital critical aortic stenosis and hypoplastic left heart syndrome (HLHS) from curative anatomical biventricular surgical repair. Surgical resection is currently the only available therapeutic option, but EFE often recurs, sometimes with an even more infiltrative growth pattern into the adjacent myocardium.
To better understand the underlying mechanisms of EFE and to explore therapeutic strategies, an animal model suitable for preclinical testing was developed. The animal model takes into consideration that EFE is a disease of the immature heart and is associated with flow disturbances, as supported by clinical observations. Thus, the heterotopic heart transplantation of neonatal rat donor hearts is the basis for this model.
A neonatal rat heart is transplanted into an adolescent rat&#39;s abdomen and connected to the recipient&#39;s infrarenal aorta and inferior vena cava. While perfusion of the coronary arteries preserves the viability of the donor heart, flow stagnation within the LV induces EFE growth in the very immature heart. The underlying mechanism of EFE formation is the transition of endocardial endothelial cells to mesenchymal cells (EndMT), which is a well-described mechanism of early embryonic development of the valves and septa but also the leading cause of fibrosis in heart failure. EFE formation can be macroscopically observed within days after transplantation. Transabdominal echocardiography is used to monitor the graft viability, contractility, and the patency of the anastomoses. Following euthanasia, the EFE tissue is harvested, and it shows the same histopathological characteristics as human EFE tissue from HLHS patients.
This in vivo model allows for studying the mechanisms of EFE development in the heart and testing treatment options to prevent this pathological tissue formation and provides the opportunity for a more generalized examination of EndMT-induced fibrosis.

著者スポットライト:内皮から間葉への移行の研究のための新生児ヘテロトピックラット心臓移植モデル  

This work presents an animal model of endothelial-to-mesenchymal transition-induced fibrosis, as seen in congenital cardiac defects such as critical aortic stenosis or hypoplastic left heart syndrome, which allows for detailed histological tissue evaluation, the identification of regulatory signaling pathways, and the testing of treatment options.

内皮から間葉への移行研究のための新生児異所性ラット心臓移植モデル

Endocardial fibroelastosis (EFE), defined by subendocardial tissue accumulation, has major impacts on the development of the left ventricle (LV) and ...