Name: re-arterialized rat partial liver transplantation with an in vivo vessel-oriented 70% hepatectomy
Uploaded: 2018-04-08T13:00:00.000+00:00
Duration: 14 min 52 s
Description: Watch this Scientific Journal Video about Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy at JoVE.com

この研究は、国家自然科学基金、中国の (許可番号 81501382) と中国浙江省地方自然科学財団のグラント号によって支持されました。LQ13H100003 および LQ16H100002)。

<ol>
	<li>Yan, J. Q., Becker, T., Peng, C. H., Li, H. W., Klempnauer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Split+liver+transplantation:+a+reliable+approach+to+expand+donor+pool.">Split liver transplantation: a reliable approach to expand donor pool.</a> Hepatobiliary Pancreat Dis Int. 4, 339-344 (2005).</li><li>Xia, R., Emond, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+partial+liver+transplantation+in+the+rat:+a+model+of+70%+hepatectomy+and+reduced+size+liver+transplantation.">Orthotopic partial liver transplantation in the rat: a model of 70% hepatectomy and reduced size liver transplantation.</a> Transplantation. 56, 1041-1043 (1993).</li><li>Omura, T., Ascher, N. L., Emond, J. 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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+procedures+for+a+rat+model+of+partial+orthotopic+liver+transplantation+with+hepatic+arterial+reconstruction.">Surgical procedures for a rat model of partial orthotopic liver transplantation with hepatic arterial reconstruction.</a> J Vis Exp. , e4376 (2013).</li><li>Sun, J., Qin, G., Wu, L., Wang, C., Sheil, A. 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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+hepatic+rearterialization+on+reperfusion+injury+and+outcome+after+mouse+liver+transplantation.">Impact of hepatic rearterialization on reperfusion injury and outcome after mouse liver transplantation.</a> Transplantation. 76, 327-332 (2003).</li><li>Huang, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+end-to-side+anastomosis+technique+for+hepatic+rearterialization+in+rat+orthotopic+liver+transplantation+to+accommodate+size+mismatches+between+vessels.">A novel end-to-side anastomosis technique for hepatic rearterialization in rat orthotopic liver transplantation to accommodate size mismatches between vessels.</a> Eur Surg Res. 47, 53-62 (2011).</li><li>Bernal, 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=Guidelines+for+rodent+survival+surgery.">Guidelines for rodent survival surgery.</a> J.Invest Surg. 22, 445-451 (2009).</li><li>Johnson, A. J., Stulting, R. D., Macsai, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Ophthalmic Microsurgical Suturing Techniques. , 21-28 (2007).</li><li>Delriviere, L., 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=Detailed+modified+technique+for+safer+harvesting+and+preparation+of+liver+graft+in+the+rat.">Detailed modified technique for safer harvesting and preparation of liver graft in the rat.</a> Microsurgery. 17, 690-696 (1996).</li><li>Oldani, G., Lacotte, S., Morel, P., Mentha, G., Toso, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+rats.">Orthotopic liver transplantation in rats.</a> J Vis Exp. , (2012).</li><li>Delriviere, L., 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=Technical+details+for+safer+venous+and+biliary+anastomoses+for+liver+transplantation+in+the+rat.">Technical details for safer venous and biliary anastomoses for liver transplantation in the rat.</a> Microsurgery. 18, 12-18 (1998).</li><li>Jin, H., 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=Preliminary+experience+of+a+PDCA-cycle+and+quality+management+based+training+curriculum+for+rat+liver+transplantation.">Preliminary experience of a PDCA-cycle and quality management based training curriculum for rat liver transplantation.</a> J Surg Res. 176, 409-422 (2012).</li><li>Selzner, N., Selzner, M., Tian, Y., Kadry, Z., Clavien, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cold+ischemia+decreases+liver+regeneration+after+partial+liver+transplantation+in+the+rat:+A+TNF-alpha/IL-6-dependent+mechanism.">Cold ischemia decreases liver regeneration after partial liver transplantation in the rat: A TNF-alpha/IL-6-dependent mechanism.</a> Hepatology. 36, 812-818 (2002).</li><li>Tanaka, H., Hashizume, K., Enosawa, S., Suzuki, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Successful+transplantation+of+a+20%+partial+liver+graft+in+rats:+a+technical+innovation.">Successful transplantation of a 20% partial liver graft in rats: a technical innovation.</a> J Surg Res. 110, 409-412 (2003).</li><li>Bockhorn, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Erythropoietin+treatment+improves+liver+regeneration+and+survival+in+rat+models+of+extended+liver+resection+and+living+donor+liver+transplantation.">Erythropoietin treatment improves liver regeneration and survival in rat models of extended liver resection and living donor liver transplantation.</a> Transplantation. 86, 1578-1585 (2008).</li><li>Martins, P. N., Neuhaus, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+anatomy+of+the+liver,+hepatic+vasculature+and+bile+ducts+in+the+rat.">Surgical anatomy of the liver, hepatic vasculature and bile ducts in the rat.</a> Liver Int. 27, 384-392 (2007).</li><li>Kamada, N., Calne, R. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+the+rat.+Technique+using+cuff+for+portal+vein+anastomosis+and+biliary+drainage.">Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage.</a> Transplantation. 28, 47-50 (1979).</li><li>Kashfi, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+various+techniques+of+orthotopic+liver+transplantation+in+the+rat.">A review of various techniques of orthotopic liver transplantation in the rat.</a> Transplant.Proc. 37, 185-188 (2005).</li><li>Wain, R. A. J., Hammond, D., McPhillips, M., Whitty, J. P. M., Ahmed, W., Ahmed, W., Jackson, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Surgical Tools and Medical Devices. , 545-562 (2016).</li><li>Huang, H., 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=Hepatic+arterial+perfusion+is+essential+for+the+spontaneous+recovery+from+focal+hepatic+venous+outflow+obstruction+in+rats.">Hepatic arterial perfusion is essential for the spontaneous recovery from focal hepatic venous outflow obstruction in rats.</a> Am J Transplant. 11, 2342-2352 (2011).</li><li>Czigany, Z., 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=Improving+Research+Practice+in+Rat+Orthotopic+and+Partial+Orthotopic+Liver+Transplantation:+A+Review,+Recommendation,+and+Publication+Guide.">Improving Research Practice in Rat Orthotopic and Partial Orthotopic Liver Transplantation: A Review, Recommendation, and Publication Guide.</a> Eur Surg Res. 55, 119-138 (2015).</li></ol>

すべての著者は開示する利害の対立があります。

ラット PLTx は、コストの高され、長い期間20トレーニング プログラムの洗練された顕微鏡視下の手順です。研究者をこの動物モデルを用いたラット PLTx プロトコルの複雑さが発生しました。フルサイズ ラット LTx と比べると、ラット PLTx 提案、微小血管移植手順の課題だけでなく、小動物の肝切除術の課題とします。したがって、詳細に全体の手順を記述する可視化顕微鏡?...

現在、寄付された肝臓器官の数と寄付された肝臓を待っている患者の数に大きな違いがあります。肝臓の臓器の不足は、地球規模の問題です。ドナー プールを展開し、分割肝移植 (LTx) および生きているドナー LTx に部分肝を移植1として使用する開発されました。
部分肝移植 (PLTx) の背後にあるメカニズムをさらに調査するには、適切な動物モデルが確立された2,3,4,5をされています。ラット PLTx、肝葉切除体内と体外している生体臓器提供者に対しての条件を模倣し、ひとそれぞれに対してを分割可視化実験のジャーナルに掲載された論文は、50% のラットを使用して、前のヴィヴォ肝5PLTx を含む詳細なプロトコルを提示しました。しかし、体内肝切除ラット PLTx はまだ報告されていない可視化文学。
体内と体外肝切除の違いに加えて自体肝切除を行う手法は、またラット PTLx を使用して多くの外科研究の現在、PLTx の結果を決定する上で重要な役割を果たしています。モデル、肝葉切除肝葉2,3,6,7,8,9椎弓根で単結紮を配置した後。ただし、切除は適していませんとして異なる肝葉のすべての肝葉の前に単結紮を置く異なった形およびサイズがあります。中央の葉の基部に単結紮部分肝移植10,11の流出に影響を与える可能性があります最終的に上大静脈の圧迫感の原因のハイリスクが伴います。したがって、ラット肝の解剖学の知識に基づいて更新肝切除法は、ラット PTLx のフィールドに必要です。
本研究では記述されているプロトコル、ラット PLTx のプロシージャに更新の容器型 70% 切除を設立されました。ポータル静脈と左葉外側 (LLL) と中葉 (ML) の肝動脈解剖、肝実質の除去の前に個別に分かれています。その後、縫合をピアスで LLL と ML の肝静脈を結紮されました。単結紮を配置するのではなく、個々 をおよび複数のピアス縫合糸を使用して、ML の残切り株上大静脈に広がることができた。したがって、外科的結紮による infrahepatic 上大静脈の収縮が回避されました。さらに、個々 を前に肝実質の除去実験11に血損失の影響を最小限に抑えること、残肝の出血率の減少によって LLL と ML の血液供給の閉塞。
それは、microsurgeons、この船の非常に小さい直径のため肝移植の適切な肝動脈 (PHA) を再構築するための重要な課題です。肝動脈を再構築するための多数の顕微鏡技術が提案された14 をされている LTx で再動脈化術が本当に必要かどうかの質問は、まだ議論12,13,14下ですが、.ここでは、最後に側に拡大 pha (CHA) 総肝動脈の吻合肝臓の移植の再本来の手法を紹介します。この辺にテクニックを使用して、2 つの肝動脈は径の大きな吻合と接続されます。吻合部の直径が大きくなるほど、実行することです簡単に手縫合より吻合部の開存の改善になります。

<table><tbody><tr><td>Surgical microscope</td><td>M&amp;ouml;ller-Wedel, Germany</td><td>654359</td><td></td></tr><tr><td>Light source</td><td>OSRAM (China) Lighting Co., Ltd</td><td>64627</td><td></td></tr><tr><td>Isoflurane Vaporizer</td><td>MIDMARK Corporation</td><td>VIP3000</td><td></td></tr><tr><td>Isoflurane</td><td>Baxter Healthcare Corporation</td><td>40032609</td><td>For Inhalation anesthesia</td></tr><tr><td>Normal Saline</td><td>Zhejiang Tianrui Pharmaceutical Co., Ltd.</td><td>716092103</td><td></td></tr><tr><td>Povidone Iodine Solution</td><td>HANGZHOU MINSHENG PHARMACEUTICAL</td><td>H33021567</td><td>For disinfection</td></tr><tr><td>Hemostatic forceps</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>J31020</td><td></td></tr><tr><td>Surgical scissors</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>JC2116</td><td></td></tr><tr><td>Needle-holder</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>J32030</td><td></td></tr><tr><td>Dressing Forceps</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>J42020</td><td></td></tr><tr><td>Mosquito hemostatic forceps</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>W40340</td><td>For liver resection; For retracting the stay sutures.&amp;nbsp;</td></tr><tr><td>Micro-scissors</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>WA1040</td><td></td></tr><tr><td>Micro needle-holder</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>WA2060</td><td></td></tr><tr><td>Curved micro typing forceps</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>WA3061</td><td></td></tr><tr><td>straight micro typing forceps</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>WA3060</td><td></td></tr><tr><td>Micro hemostatic forceps</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>WA3020</td><td></td></tr><tr><td>Bulldog clamp</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>XEC350</td><td></td></tr><tr><td>Vessel clamps</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>W40160</td><td></td></tr><tr><td>Micro Serrefine-curved</td><td>&amp;nbsp;Fine Science Tools, Inc</td><td>18055-05</td><td></td></tr><tr><td>Dacryosyrinx</td><td>Shi Zhuo medical instruments Co., Ltd.</td><td>5# curve</td><td>Use as the curved needle for forcing out the air bubbles in the vascular lumen</td></tr><tr><td>Sterilized gauze swabs</td><td>Fuqing Health &amp;amp; Integral Medical</td><td>20230R</td><td></td></tr><tr><td>5mL syringe</td><td>Zhejiang Yusheng Medical Instrument Co.,Ltd</td><td>811932416</td><td></td></tr><tr><td>18G needles</td><td>Shanghai Kindly Enterprise Development Group Co., Ltd</td><td>01-014</td><td>For fixing the upper abdomen wall</td></tr><tr><td>Intima II 22G intraveous catheter</td><td>Becton Dickinson Medical Devices (Shanghai) Co Ltd.</td><td>383207y</td><td>For making biliary stent; For infusing perfusate</td></tr><tr><td>7-0 polypropylene sutures</td><td>Ningbo Medical Needle Co., Ltd</td><td>151026</td><td>For SHVC anastomsis</td></tr><tr><td>6-0 silk suture</td><td>Shanghai Pudong Jinhuan Medical Products Co., Ltd</td><td>2650182</td><td>For ligation</td></tr><tr><td>Culture dish (100mm)</td><td>Corning Incorporated</td><td>430165</td><td>used in back table for storage ice. The back table dish was placed on it.&amp;nbsp;</td></tr><tr><td>Culture dish (30 mm)</td><td>Corning Incorporated</td><td>3296</td><td>As a vessel for organ preservation</td></tr><tr><td>Angiocath 12G intravenous catheter</td><td>Becton Dickinson Medical Devices (Shanghai) Co Ltd.</td><td>382277</td><td>For making a IHVC cuff</td></tr><tr><td>Angiocath 14G intravenous catheter</td><td>Becton Dickinson Medical Devices (Shanghai) Co Ltd.</td><td>382269</td><td>For making a PV cuff</td></tr><tr><td>Buprenorphine (hydrochloride)</td><td>Tianjin Institute of Pharmaceutical Research Pharmaceutical Co., Ltd</td><td>H12020275</td><td>For analgesic</td></tr><tr><td>Cefuroxime sodium for injection</td><td>Esseti FarmaceuticiS.r.l</td><td>H20160013</td><td>As an antibiotics</td></tr></tbody></table>

re-arterialized rat partial liver transplantation with an in vivo vessel-oriented 70% hepatectomy

分割肝移植と生体肝ドナー肝移植肝臓臓器をより効率的に活用する診療所で開発されました。これらの手術の背後にあるメカニズムを理解、関連する外科研究ラット部分肝移植 (PLTx) モデルを設置しました。ラットの PLTx モデルの複雑さのため、詳細な説明とプロトコルが必要です。掲載された記事は以前体外肝切除が 50% ラット PLTx を達成するために使用されたプロトコルを報告しました。このプロトコルとは対照的、生体内で70% 切除再門脈 PLTx を導入しました。更新された容器指向の肝切除は、顕微鏡のプロシージャを調整する PLTx のラットに組み込まれました。ポータル静脈と左葉外側と中央の葉の肝動脈が個別に解剖し、残肝切り株で出血の確率を減らす、肝実質の除去前に結紮.さらに、拡大固有肝動脈と総肝動脈側に血管吻合は、肝動脈を再 arterialize に導入されました。この側に血管吻合技術を用いたは、手縫合の難しさを減少し、吻合部の開存性の高率を維持、吻合部の直径が拡大されました。また、infrahepatic 下大静脈のカフ吻合は若干変更されました。受信者の下大静脈周囲円周の肝実質のセクションは、カフ吻合血管の内腔の三次元形状を維持するために中に保たれていた.肝実質のこのセクションは、吻合部を完了した後削除されました。この変更を加えたカフ吻合時間をさらに短縮宿泊縫合の配置を含む工程は省略されました。ラット PLTx のこのプロトコルを使用して、手術後肝酵素の低レベル、そのまま肝小葉アーキテクチャと高い生存率が達成されました。

合計では、ラット門脈 PLTx 31 例は、このプロトコルを使用して完成しました。すべての受信者は、観測時間の最後まで生き残った。受信者の体重は術後 (POD) 4 後回復し始めた。体重の斜面は、間接的に受信者の回復ポッド 6 (図 8) 後、ルイス ・ ラットが正常に近いでした。組織学的胆管のわずかな増殖を認めた、受信者と受信者の肝小葉アーキ...

Watch this Scientific Journal Video about Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy at JoVE.com

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

ここでは、再門脈ラット部分肝移植を含むプロトコルが表示されます。具体的には、70% 肝切除だった体内肝の血管指向の更新手法を用いた肝動脈は、-側の方法で再建されました。カフのテクニックは、infrahepatic 下大静脈の吻合時間を短縮に変更されました。

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. ...

re-arterialized-rat-partial-liver-transplantation-with-an-vivo-vessel

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

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

11.5K ビュー.  The First Affiliated Hospital of Wenzhou Medical University. ここでは、再門脈ラット部分肝移植を含むプロトコルが表示されます。具体的には、70% 肝切除だった体内肝の血管指向の更新手法を用いた肝動脈は、-側の方法で再建されました。カフのテクニックは、infrahepatic 下大静脈の吻合時間を短縮に変更されました。

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

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 ...

Visualization of Vascular and Parenchymal Regeneration after 70% Partial Hepatectomy in Normal Mice

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the silicone compound, via a 26 G catheter, into the portal or hepatic vein. After silicone injection, organs were explanted and prepared for ex-vivo micro-CT (&#181;CT) scanning. The silicone injection procedure is technically challenging. Achieving a successful outcome requires extensive microsurgical experience from the surgeon. One of the challenges of this procedure involves determining the adequate perfusion rate for the silicone compound. The perfusion rate for the silicone compound needs to be defined based on the hemodynamic of the vascular system of interest. Inappropriate perfusion rate can lead to an incomplete perfusion, artificial dilation and rupturing of vascular trees.
The 3D reconstruction of the vascular system was based on CT scans and was achieved using preclinical software such as HepaVision. The quality of the reconstructed vascular tree was directly related to the quality of silicone perfusion. Subsequently computed vascular parameters indicative of vascular growth, such as total vascular volume, were calculated based on the vascular reconstructions. Contrasting the vascular tree with silicone allowed for subsequent histological work-up of the specimen after &#181;CT scanning. The specimen can be subjected to serial sectioning, histological analysis and whole slide scanning, and thereafter to 3D reconstruction of the vascular trees based on histological images. This is the prerequisite for the detection of molecular events and their distribution with respect to the vascular tree. This modified silicone injection procedure can also be used to visualize and reconstruct the vascular systems of other organs. This technique has the potential to be extensively applied to studies concerning vascular anatomy and growth in various animal and disease models.

Tools used for visualizing vascular regeneration require methods for contrasting the vascular trees. This film demonstrated a delicate injection technique used to achieve optimal contrasting of the vascular trees and illustrate the potential benefits resulting from a detailed analysis of the resulting specimen using &#181;CT and histological serial sections.

正常マウスで70％肝切除後の血管と実質再生の可視化

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the ...

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 ...

Intrasplenic Transplantation of Hepatocytes After Partial Hepatectomy in NOD.SCID Mice

Partial hepatectomy is a versatile and reproducible method to study liver regeneration and the effect of cell based therapeutics in various pathological conditions. Partial hepatectomy also facilitates the increased engraftment and proliferation of transplanted cells by accelerating neovascularization and cell migration towards the liver. Here, we describe a simple protocol for performing 30% hepatectomy and transplantation of cells in the spleen of a non-obese diabetic/severe combined immunodeficient NOD.SCID (NOD.CB17-Prkdcscid/J) mouse. 
In this procedure, two small incisions are made. The first incision is to expose and resect the left lobe of the liver, and another small incision is made to expose the spleen for the intrasplenic transplantation of cells. This procedure does not require any specialized surgical skills, and it can be completed in 5-7 minutes with less stress and pain, faster recovery, and better survival. We have demonstrated the transplantation of hepatocytes isolated from a green fluorescent protein (GFP) expressing mouse (Transgenic C57BL/6-Tg (UBC-GFP) 30Scha/J), as well as hepatocyte like cells of human origin (NeoHep) in partially hepatectomized NOD.SCID mice.

We have described a protocol for performing partial hepatectomy (PHx) and cell transplantation via spleen in NOD.SCID (NOD.CB17-Prkdcscid/J) mice. In this protocol, an incision is made to expose and resect the left lobe of the liver followed by another incision for the intrasplenic transplantation of cells.

うなずくにおける肝切除後肝脾内移植。SCID マウス

Partial hepatectomy is a versatile and reproducible method to study liver regeneration and the effect of cell based therapeutics in various pathological ...

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

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep learning curve. The introduction of the cuff technique for anastomosis of the portal vein (PV) and infrahepatic vena cava (IHVC) has significantly simplified the transplant procedure in rats. However, due to the short anterior wall of the recipients' suprahepatic vena cava (SHVC), the cuff technique is very difficult to use for the reconstruction of the SHVC. Most researchers in this field still use the hand-suture technique for SHVC reconstruction, which makes it the bottleneck step in rat orthotopic liver transplantation. The magnetic anastomosis technique (i.e., magnamosis) is a method of connecting two vessels using the attractive force between two magnets. Our recent study has shown that the magnetic anastomosis technique is superior to the hand-suture technique for SHVC reconstruction in rats. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using the novel magnetic anastomosis technique. In this model, the reconstruction of the PV and IHVC was performed by the standard cuff technique, while the reconstruction of the bile duct (BD) was performed by a stent technique. The hepatic re-arterialization was not performed. The magnetic anastomosis technique made SHVC reconstruction much easier and significantly shortened the anphepatic phase. After a reasonable learning curve, even researchers without advanced microsurgical skills can produce reliable and reproducible results using this rat model of OLT.

The reconstruction of the suprahepatic vena cava (SHVC) remains a difficult step in rat orthotopic liver transplantation. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using a novel magnetic anastomosis technique.

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 ...

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 ...

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 ...

<meta charset="utf8"></head><body>ラットにおける肝移植と同種移植片の開存性の分析

Heterotopic Auxiliary Whole Liver Rat Transplant Model Utilizing a Hepaticoureterostomy for Allograft Rejection Studies

Small animal transplant models are indispensable for organ tolerance studies investigating feasible therapeutic interventions in preclinical studies. Rat liver transplantation (LTx) protocols typically use an orthotopic model where the recipients&#39; native liver is removed and replaced with a donor liver. This technically demanding surgical procedure requires advanced micro-surgical skills and is further complicated by lengthy anhepatic and lower body ischemia times. This prompted the development of a less complicated heterotopic method that can be performed faster with no anhepatic or lower body ischemia time, reducing post-surgery stress for the recipient animal.
This heterotopic LTx protocol includes two main steps: excising the liver from the donor rat and transplanting the whole liver into the recipient rat. During the excision of the donor liver, the surgeon ligates the supra-hepatic vena cava (SHVC) and hepatic artery (HA). On the recipient side, the surgeon removes the left kidney and positions the donor liver with the portal vein (PV), infra-hepatic vena cava (IHVC), and bile duct facing the renal vessels. Further, the surgeon anastomoses the recipient&#39;s renal vein end to end with the IHVC of the liver and arterializes the PV with the renal artery using a stent. A hepaticoureterostomy is utilized for biliary drainage by anastomosing the bile duct to the recipient&#39;s ureter, permitting the discharge of bile via the bladder.
The average duration of the transplantation was 130 min, cold ischemia duration was around 35 min, and warm ischemia duration was less than 25 min. Hematoxylin and eosin histology of the auxiliary liver from syngeneic transplants showed normal hepatocyte structure with no significant parenchymal alterations 30 days post-transplant. In contrast, 8-day post-transplant allogeneic graft specimens demonstrated extensive lymphocytic infiltration with a Banff Schema rejection activity index score of 9. Therefore, this LTx method facilitates a low morbidity rejection model alternative to orthotopic LTx.

<meta charset="utf8"></head><body>同種移植拒絶反応研究のための肝瘻造設術を用いたヘテロトピア補助全肝ラット移植モデル

The rat heterotopic auxiliary liver transplant protocol described here offers a practical investigational tool for exploring mechanisms of hepatic allograft rejection. This model helps to alleviate the surgical hurdles and animal stress of orthotopic liver transplantation in rats.

同種移植拒絶反応研究のための肝瘻造設術を用いたヘテロトピア補助全肝ラット移植モデル

Small animal transplant models are indispensable for organ tolerance studies investigating feasible therapeutic interventions in preclinical studies. Rat ...