Bu çalışma &quot;Deutsche Forschungsgemeinschaft&quot; (DFG) tarafından finanse edildi. K. Schleimer NRW Lise Meitner Bilim Bakanlığı Bursu ve Araştırma tarafından desteklenmiştir. Biz onu mükemmel düzenleme yardım için Bayan Mary-Joan Blümich teşekkür etmek istiyorum. Dr Maria Kokozidou Fresenius 2011_A61 finanse edildi.

1. Deney Hayvanları <ol><li> Hayvan koruma FELASA kurallarına uygun olarak, yapılacaktır tüm deneyler için bir Yerel Etik Kurul onayı (K 40, 17/99) edinin. </li><li> (Örneğin, Charles River WIGA GmbH, Sulzfeld, Almanya) herhangi bir ticari üreticiden satın almak için erkek Lewis sıçanları mevcuttur kullanın. Burada, 224 hayvanlar (112 bağış, 112 alıcılar) 112 nakli gerçekleştirmek için kullanılmıştır. Alıcılar, ameliyat sonrası 6 haftada feda edildi. Operasyon sırasında, hayvanlar 250-350 g ağırlıkta olması gerekmektedir. </li><li> Yiyecek ve su ücretsiz erişim ile iklim kontrollü odalarda hayvanları tutun. </li></ol> 2. Anestezi ve Operatif Hazırlık <ol><li> 1 l / dk arasında bir oksijen akış hızı ve anestezi sağlamak için% 5 anestezi gazı ile bir buharlaştırıcı izofloran ile anestezi makineye bağlı bir pleksiglas kutuya sıçan yerleştirin. % 1.5 isofluorane de anestezi koruyun. </li><li> Yılında sıçan yerleştirinBir ısınma plaka üzerinde sırtüstü yatar pozisyonda. </li><li> Bir epilasyon kremi ile karın kürk çıkarın. </li><li> Aşağıdaki gibi steril bir cerrahi alan hazırlayın: benchtop temizlemek için bir dezenfektan kullanın. Ilk kesi sitesi kullanım klorheksidin dezenfeksiyon, alkol saniye ve son olarak, bodur solüsyonu (örn, klorheksidin ve% 70 izopropil alkol) için. Bir tek kullanımlık cerrahi önlük ve cerrahi eldiven giyin. </li><li> Steril mikrocerrahi aletler kullanın. </li><li> 12-20 kat büyütmeli bir operasyon mikroskobu kullanın. </li></ol> 3. Cerrahi Prosedür <ol><li> Donör operasyonu Hazırlık <ol><li> Median laparotomi gerçekleştirin ve sol lateral lob ve Higgins tarafından açıklandığı gibi greft medyan lobu rezeke. 14 </li><li> Daha sonra triadın hazırlar. İlk küçük safra tıkanıklığı ve dilatasyon uyarmak için safra Arter. Daha sonra bağlanır ve gastroduode transeksiyonunal ven ve arter. Daha sonra, çölyak gövde ve ortak hepatik arter izole ve sol gastrik arter ve dalak arteri transect. Sıkışık safra kanalı kesilirken, ve içine bir 20-G stent tanıtmak. Stent yerleştirme bütün hilar safra kanalı sistemi drenaj sağlamalıdır. Stent distal safra transect. </li><li> Sağ sağ renal venin üstüne kaval ven infrahepatic segmenti izole eder. Perfüzyon </li><li> Çölyak gövde üzerinde aorta bağlanmasını takiben, portal ven kelepçe ve sarın. Portal ven içine perfüzyon kateter (14 g) ortaya koyar ve histidin-triptofan-ketoglutarat (HTK) solüsyonunun 30 ml (12 cm H2O, 4 ° C) ile perfüzyon gerçekleştirin. </li><li> Diyaframı ve perfüzata dışarı akmasına izin suprahepatik kaval ven kesti. </li><li> Aort yama ile birlikte çölyak gövde tüketim ve histidin-triptofan-ketoglutarat çözümü ile durulayın. Eksplantasyonu </li><li> Sağ sağ renal venin yukarıda infrahepatic kaval ven transect. </li><li> Ardından, portal vende 0,3 mm çapında bir stent yerleştirmek ve dikilen ile sabitleyin. Aşağıdaki işlemi sırasında, portal ven sağ renal arter yoluyla arteriyelize edilir. Stent distal portal ven transect. </li><li> Suprahepatik kaval ven ligasyonu sonra, diyaframın bir manşet ile birlikte transect. </li><li> Greft Eksplantasyona ve buz üzerinde bir plastik torba içinde 4 ° santigrat histidin-triptofan-ketoglutarat (HTK) solüsyonunun saklayın. Infrahepatic kaval ven yerleştirin köşe dikişler (8-0 etilon). </li></ol></li><li> Alıcı operasyonu <ol><li> Median laparotomi gerçekleştirin ve infrahepatic kaval ven hazırlamak. Ardından, sağ renal arter ve üreter izole üreter Arter ve transect. Kökeni de sağ renal arter Kelepçe, sonra sağ renal ven Arter. Sonra her iki kesilerinden sonra yenidennal gemiler, nefrektomi yapma. Daha sonra, sağ renal arter kısaltmak heparin tuzlu su çözeltisi (40 UI heparin / ml tuz çözeltisi) ile durulayın. </li><li> Infrahepatic kaval ven oval lümen oluşturmak için infrahepatic kaval ven proksimal ve distal klemplemesinden sonra dikilen proksimal sağ renal ven transect. Heparin tuzlu su çözeltisi ile kaval ven durulayın. Alıcının karnın sağ renal bölgede greft dikiniz. </li><li> Bir 8-0 etilon çalışan sütür ile Alıcının kaval ven donörün infrahepatic kaval ven uç-yan anastomoz. </li><li> Heparin / tuzlu su çözeltisi ile yıkandıktan sonra, sağ renal arter (Şekil 3) içine portal ven stent getirmektedir. 2 ilmek (8-0 etilon) ile portal ven ve renal arter bağlayın. Bir bağ (örgülü ipek siyah 7-0) ile renal arter bağlayan stent Fix. </li><li> Reperfüzyona izin verecek şekilde, ilk olarak, daha sonra, arteryel kelepçe kaldırmakvenöz kelepçeleri (Şekil 4). </li><li> Infrarenal aort hazırlayın, sonra yakın ve uzak microclamp yerleştirin ve sağ hem de kelepçeler arasındaki aort insizyon. Bir 10-0 etilon çalışan sütür ile, aort yama kullanarak, aorta ve vericinin çölyak gövde arasında bir uç-yan anastomoz gerçekleştirin. </li><li> Duodenum içine safra kanalı yerleştirin: duodenum duvarında bir kese dikişi uygulayın, sağ kese ortasında duodenum duvarında küçük bir kesi yapmak. Duodenum içine stent safra kanalı tanıtmak ve kese dikişi sıkın. Son olarak, sıkıca birbirine iki dikiş (etilon 8-0) (Şekil 5) ile onikiparmak bağırsağı ve safra kanalı tutun. </li><li> Daha sonra doğal karaciğer rezeksiyonu ara toplam birinci Emond ve diğerleri tarafından tarif gerçekleştirin. Akut karaciğer yetmezliği taklit. 7 </li><li> Yeniden lateral karın duvarına, birlikte diyafram bir manşet ile, greft suprahepatik kaval ven Fix4-0 vicryl sütür ile cipient. Tuzlu su çözeltisi (Şekil 6) ile karın durulayın. </li><li> Kas tabakası için 4-0 vicryl çalışan dikiş ve kutanöz için 3-0 vicryl çalışan dikiş kullanın. </li></ol></li></ol> 4. Kurtarma ve Postoperatif Yönetimi <ol><li> Hayvanlar bir kızılötesi ısınma lambası altında anestezi kurtarmak edelim. </li><li> 2 gün boyunca 0,03-0,05 mg / kg bw sn her 8-12 st bir dozda buprenorfin enjekte edilir. </li><li> Genel durumunu ve hastalık ya da enfeksiyon belirtileri ilişkin hayvanları inceleyin. </li></ol> İlk Yerli Karaciğer Ağırlık 5. Hesaplama Aşağıdaki şekilde, ilk ana karaciğer ağırlığı hesaplayın: Tüm karaciğer ağırlığı rezeke karaciğer lobları ağırlığını çıkarmak içinden (vücut ağırlığının (+/-% 0,49 arasında bir standart sapma ile bu değer% 2.66, N ile önceki deneylerde saptandı = Aynı soy ve a 80 hayvanlarge)). 15

<ol>
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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. 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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. 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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>

Tüm Yazarlar herhangi bir çıkar ilan etti.

İlk heterotopik yardımcı sıçan karaciğer nakli Lee ve arkadaşları tarafından tanımlanmıştır., 1966 yılında 16 Hess tarafından izledi. Greft 17 portal ven alıcının portal ven ile anastomoz edildi. Bu işlem sırasında, yerli karaciğer portal venöz kan dolaşımı tehlikeye düşer. 1991 yılında Hong kan akış regülasyonu büyük portal olmadan. Alıcının iliak arter yoluyla PVA ile 13 DURDURMASI gerçekleştirilen ...

Akut karaciğer yetmezliği% 60-80 mortalite oranı. 8 akut karaciğer yetmezliği yol açan klinik tıpta önemli bir sorun teşkil etmektedir, yardımcı karaciğer transplantasyonu akut faz köprü için umut verici bir alternatif tedavi yöntemidir. Yerli karaciğer immünosüpresyonun ilaçlar kesilebilir rejenerasyon aşağıdaki yenilemek için sol ve. Bunun bir sonucu olarak, aşı Atrofik olacaktır. Ancak yardımcı karaciğer nakli için ameliyat tekniği halen tartışılmaktadır. 1-3 Bu bağlamda, Terpstra greft için alan eksikliği rapor, portal kan fenomen ve venöz konjesyon ve drenaj sorunu çalmak. 9-10 donörün portal ven Alıcının portal ven uca-yan anastomoz edildi. Bu greft ve yerli karaciğer arasında ve portal ven trombozu portal akım rekabet sonuçlandı. Bu teknik sorunların çoğu APOLT-yöntemin kullanılması (orthotop ile önlenebiliric yardımcı segmentleri implantasyonu). 4 Ancak bu yöntem doğal karaciğer ve greft. 4 1998 yılında hem geniş rezeksiyon gerektirir, Erhard ve ekibi portal ven ve arteri (PVA) (Şekil 1 kullanan heterotopik yardımcı karaciğer transplantasyonu (HALT) geliştirdi ). Hilusu ve yerli karaciğerin portal ven bu yöntemin kullanımı ile saygı gösterilir. Bu yerli karaciğer kurtarma rahatsız olabilir gibi portal ven trombozu ve portal kan gibi istenmeyen yan etkileri alıcı çalmak fenomen, yedek parça. Buna ek olarak, doğal karaciğer ve greft yaygın bir biçimde karaciğer rezeksiyonu (greft) ve rejenerasyon (yerli karaciğer) kapatmak için kullanılabilecek ve sonuçta daha az kanama ve daha fazla kalan toplam karaciğer parenkimal kütlesi ile, bu yöntem için gereklidir. Bu teknik ilk klinik sonuçlar vaat gösterdi. 5-6 Bu deneysel çalışmanın amacı portal ven ve arteri içinde olan bir HALT tekniğinin gelişmesi olduAraştırma amacıyla, sıçan (Şekil 2). Ilk deneylerde karaciğer kapsül reperfüzyon ve masif kanama sonrasında nakledilen greft tıkanıklık sonuçlandı, damar dikkate arteriyelize portal kan akımı regülasyonu sürmedi. 11 arteriyelize portal vende sonra kan akışı bu yana 0.3 mm&#39;lik bir iç çapa ve 8 mm arasında bir uzunluğa sahip olan bir stent ile ayarlanmıştır. Pilot Deneyler, 0.3 mm&#39;lik bir stentin çapı, üst fizyolojik aralığında arteriyelize portal ven içinde bir ortalama kan akımını verir olduğunu göstermiştir (1.7 +/- 0.4 ml / dak / g karaciğer ağırlık). Kan akımı düzenleme özelliğine sahip 12 Sonuçlarımız kan akış regülasyonu büyük portal olmadan alıcının iliak arter yoluyla PVA ile yaptığı deneylerde. 13 DURDURMASI gerçekleştirilen ve sinüzoid kısıtlarını meydana Hong, sonuçları ile ilişkilidir. Bizim video amacı d etmektiradım adım emonstrate sıçanda bu ne.w deneysel cerrahi tekniği. Uzun vadede önerilen teknik karaciğer rejenerasyonu ve inter-karaciğer rekabet ve portal ven damarlanma, portal ven hiperperfüzyon ve portal kan akımı düzenlenmesinde çalışmalarda kullanılabilir.

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			<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

Akut karaciğer yetmezliği yardımcı karaciğer nakli ilginç bir alternatif bir yaklaşımdır. Amaç başarısız doğal karaciğer yeniden kadar geçici bir destek sağlamaktır. 1-3 APOLT metodu, yardımcı kesimleri-en ortotopik implantasyon teknik sorunların çoğunu kaçırır. Ancak bu yöntem doğal karaciğer ve greft. 4 1998 yılında hem geniş rezeksiyon gerektirir, Erhard portal ven ve arteri (PVA) (Şekil 1) kullanılarak heterotopik yardımcı karaciğer transplantasyonu (HALT) geliştirdi. Bu teknik ilk klinik sonuçlar vaat gösterdi. 5-6 Biz greft morfolojisi ve fonksiyonu (Şekil 2) debi ayarlı PVA etkisini incelemek için sıçan akış düzenlenen PVA ile, bir HALT-teknik geliştirdi. Orijinal boyutunun% 30 düşürülmüş bir karaciğer greft, heterotopically sağ böbreğin eksplantasyonu sonra alıcının sağ renal bölgede implante edildi.60; Greft kızıl-hepatik kaval ven alıcının infrahepatic kaval ven ile anastomoz edildi. Donörün portal ven arteriali stent tekniği ile alıcının sağ renal arter yoluyla yürütülmüştür. Arteriyelize portal ven kan akış düzenleme 0.3 mm&#39;lik bir iç çapa sahip bir stent kullanımı ile elde edilmiştir. Greft çölyak gövde uç-yan alıcının aorta ve safra kanalı ile anastomoz duodenum içine implante oldu. Yerli karaciğer subtotal rezeksiyon akut karaciğer yetmezliğine yol açan yapıldı. 7 Bu şekilde, 112 transplantasyon yapılmıştır. Perioperatif sağkalım oranı% 90 idi ve 6 haftalık sağkalım oranı% 80 idi. Altı hafta işleminden sonra yerli karaciğer 9.8 ± 1 g ile 2.3 ± 0.8 g ağırlığında bir artış gösteren sokulabilir. Şu anda, greft ağırlığı 2.3 ± 0.8 g 3.3 ± 0.8 g &#39;a düşmüştür. </p&gt; Biz greft morfolojisi ve fonksiyonu açısından umut verici uzun vadeli sonuçlar elde etmek mümkün. Yerli karaciğer yeniler kadar akış düzenlenen PVA ile HALT güvenilir akut karaciğer yetmezliği köprü.

Transplantasyon süresi ortalama yaklaşık 150 dakika idi. Soğuk iskemi zamanı, 64 +/- 7 dk sıcak iskemi süresi en az 25 dakika idi. Perioperatif sağkalım oranı% 90 idi ve 6 haftalık sağkalım oranı% 80 idi. % 20 kayıp postoperatif ilk haftasında çünkü stent çıkık ve safra kaçağı (dört hayvanlar) peritonit nedeniyle oldu ve nedeni aşağıdaki beş hafta boyunca pnömoni, karın içi abse ve ileus (yedi hayvanlar). Hayatta kalan hayvanların mükemmel bir genel durumda...

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

Yardımcı karaciğer nakli başarısız karaciğer rejenerasyonu kadar, akut karaciğer yetmezliğinde geçici bir destek sağlar. Portal ven ve arteri (PVA) ile heterotopik yardımcı karaciğer transplantasyonu (HALT) yeterli karaciğer fonksiyon oluşturur. Biz greft morfolojisi ve fonksiyonu üzerinde portal ven arteriali etkisini incelemek için, sıçan benzer bir tekniği geliştirdi.

Heterotopik Yardımcı Sıçan Karaciğer Nakli ile Akut Karaciğer Yetmezliği Portal Ven arteriali Akış ayarlı

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 Görüntüleme.  University Hospital RWTH Aachen.  Yardımcı karaciğer nakli başarısız karaciğer rejenerasyonu kadar, akut karaciğer yetmezliğinde geçici bir destek sağlar. Portal ven ve arteri (PVA) ile heterotopik yardımcı karaciğer transplantasyonu (HALT) yeterli karaciğer fonksiyon oluşturur. Biz greft morfolojisi ve fonksiyonu üzerinde portal ven arteriali etkisini incelemek için, sıçan benzer bir tekniği geliştirdi. 

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

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.

Sıçanlarda Ortotopik Karaciğer Nakli

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

Tıp

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.

Hepatik Arter İmar Kısmi Ortotopik Karaciğer Nakli Bir Rat Modeli Cerrahi İşlemleri

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

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.

Sıçan Heterotopik Abdominal Kalp / Bir Ses yüklenen Yapılandırması Tek akciğer transplantasyonu

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.

Farelerde Heterotopik Servikal Kalp Nakli

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

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ischemia/reperfusion (I/R) injury with myriad negative consequences. Potential I/R injury in marginal organs destined for liver transplantation contributes to the current donor shortage secondary to a decreased organ utilization rate. A significant need exists to explore hepatic I/R injury in order to mediate its impact on graft function in transplantation. Rat liver hilar clamp models are used to investigate the impact of different molecules on hepatic I/R injury. Depending on the model, these molecules have been delivered using inhalation, epidural infusion, intraperitoneal injection, intravenous administration or injection into the peripheral superior mesenteric vein. A rat liver hilar clamp model has been developed for use in studying the impact of pharmacologic molecules in ameliorating I/R injury. The described model for rat liver hilar clamp includes direct cannulation of the portal supply to the ischemic hepatic segment via a side branch of the portal vein, allowing for direct segmental hepatic delivery. Our approach is to induce ischemia in the left lateral and median lobes for 60 min, during which time the substance under study is infused. In this case, pegylated-superoxide dismutase (PEG-SOD), a free radical scavenger, is infused directly into the ischemic segment. This series of experiments demonstrates that infusion of PEG-SOD is protective against hepatic I/R injury. Advantages of this approach include direct injection of the molecule into the ischemic segment with consequent decrease in volume of distribution and reduction in systemic side effects.

A unique rat liver hilar clamp model was developed for studying the impact of pharmacologic molecules in ameliorating ischemia-reperfusion injury. This model includes direct cannulation of the portal supply to the ischemic liver segment via a branch of the portal vein, allowing for direct hepatic delivery.

Bir Sıçan Karaciğer Hilar Kelepçe Modeli Kullanılarak Doğrudan Segmental İçi hepatik Dağıtım Yöntemi

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

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.

Karaciğer bölümü ilişkilendirme ve Portal ven ligasyonu hazırlanmış Hepatectomy (ALPPS) yordamı için fare modeli

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.

Yeniden Arterialized sıçan kısmi karaciğer nakli bir vivo içinde gemi odaklı % 70 ile Hepatectomy

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.

Sirozlu fareleri Portal hipertansif sendromunda invaziv hemodinamik karakterizasyonu

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 Parsiyel Hepatektomi ve Asetaminofen birleştirerek Akut Karaciğer Yetmezliği Bir Sıçan Modeli Nin Üretimi

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.

Ortotopik Karaciğer Transplantasyonunun Sıçan Modelinde Arteriyel Reconnection Sonrası Azaltılmış Komplikasyonlar

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