저자는 기술 지원을 위해 파스칼 Paschenda과 Mareike Schulz 감사드립니다.

<ol>
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우리는 공개 할 충돌 관심이 없습니다. 쥐가 실험실 동물 과학 협회 (FELASA)에 대한 연방 정부의 지침에 따라 특정 - 병원체가없는 조건에서 자리 잡고 있습니다. 모든 실험 동물의 보호와 &quot;실험실 동물의 관리 및 사용에 대한 안내&quot;(보건 발행 번호 86-23 국립 연구소, 1985 개정)에 관한 독일 연방 법에 따라 실시되었다.

쥐 올트의 첫 번째 모델은 리 외에 의해보고되었다. 1,973 9, 간장 동맥을 포함한 모든 선박은 손 놓은 방법으로 재건되었으며 extracorporeal portosystemic 션트가 사용 된 인치 이 모델은 기술적으로 복잡하고 수행하는 것이 어려웠습니다. 다음 모델은 1975 년 10 동일한 저자에 의해 개발 된 간장 동맥 재건 및 extracorporeal 션트없이 하나했습니다. 그 후 1979 년, Kamada 외이 있습니다. 간 rearterialization 11없이 모델에 대한 커프 문합 기술을 도입했습니다. 이러한 수정을 통해 쥐의 올트받는 작업의 단축 anhepatic 시간으로 단순화 된 널리 인정 실험 모델로 사용되었습니다. arterialization가 까다로운 작업했지만하지만,이 쥐의 간 arterialization 올트 8 중요성으로 이후 많은 논란이있었습니다 디이식 후 생존에 영향을 미치지 싶네요. 다양한 재건 기술을 이용하여 간 arterialization에 대한 많은 연구가 이러한 대동맥 세그먼트 - 투 - 대동맥 문합 3,9,17, 커프 문합 기술 18,19,20, 텔레 스코핑 기법 (5), 스텐트 기술 13 등 8,보고되었습니다 16, 및 슬리브 문합 기술 12,21-23. 쥐 올트에 대한 기술이 오늘날 표준화되지 않지만, arterialized 모델은 점점 더 그 생리 우위 8,12,13,14의 관점에서 선호하고 있습니다. 위에서 언급 한 기술 중 수행 할 간단하고 빠르 스텐트 기술은 레만 외에 의해보고되었다. 2005 년 16. 연구는 우수한 결과를 보여 주었다 : 더 가림 율은 reperfusion 후 8 시간, 24 시간, 6 개월에서 복원 간장 동맥에서 관찰되지 않았습니다. 따라서 간 arterialization이 기술을 채택했다. 우리는 perforSHVC와 IHVC의 재건을위한 mA 손집니다 문합. 이 방법은 혈전증 8 감소 발생에 이르게 최적의 생리 상태와 anastomotic 사이트를 제공하고 최고의 미세 시뮬레이션 및 의사에 대한 교육입니다. 또한, 문합는 짧은 선박 stumps 가능 할 수 있습니다. IHVC의 문합에 대하여,이 방법은 커프 문합 기술에 비해 이식 측면에 긴 IHV​​C을 필요로하지 않습니다. 따라서 기증자 신장 정맥이 오래 이식 IHVC를 만들기 위해 해부를 할 때,이 방법은 짧은 intrahepatic과 오른쪽 측면과 꼬리가있는 엽으로 구성되어 30 %의 이식과 같은 긴 IHV​​C을 필요로 작은 이식​​의 이식 적용됩니다 SHVC이없는 대정맥. 여러 가지 방법을 현재까지 쥐에서 간 절제술의 기술에 관한 것이보고되었습니다, 두 가지 주요 기술은 전통적인 대량 봉합사 기술 아르그리고 선박 중심의 기술 24. 우리는 50 % 간 절제술 15 클래식 봉합사 기술을 수행하지만, 수술 현미경 절차 미세한를하고, 나머지 엽과 구조에 대한 손상을 방지 할 수 있습니다. 우리는 우리의 모델에서받는 쥐의 대표 결과를 설명하고, 쥐가 명백한 합병증없이 7 일간 관찰 기간 동안 살아 남았습니다. 모델은 같은 장기간 냉장, 심장 사후 기증을 포함 연장 따뜻한 국소 빈혈 및 간 부상 또는 질병의 실험 모델에서 작은 간 이식 또는 이식의 사용과 같은 다른 설정을 선택하여 실험의 다른 목적으로 수정할 수 있습니다. , 운영 시간, 전자 출혈의 양 : 우리의 경험에서, 이식, 쥐 올트의 결과에 대해 가장 신뢰할 수있는 매개 변수를 한 후 생존에 영향을 미칠 수있는 절차를 통해 세 가지 핵심 요소가 있습니다특히 포털 정맥과 IHVC 시간을 클램핑하고, 각 선박의 재건의 적절성은 어느 협착증, 혈전증이 발생할 수, 또는 출혈이. 이 모델의 연수 기간에서 실패의 대부분은 아마도 이러한 요인과 관련이있을 수도 있습니다. 이 비디오 문서에서는, 우리는 간장 동맥 재건과 일부 올트 우리의 쥐 모델에 대한 수술 절차에 대한 단계별 지침을 제시한다. 올트의 쥐 모델이 복잡하고 고급 microsurgical 기술을 필요로하는 반면,이 문서에서는이 모델의 교육 및 학습을위한 좋은 가이드 역할을해야 실질적인 정보를 충분히 제공합니다. 효율적으로이 모델을 학습하면 학습 기간을 단축 연습에 필요한 동물과 비용의 수를 줄이고, 나중에 실험에서 신뢰할 수있는 결과를 재현을 위해 특히 중요합니다. 이에 러셀과 버치가 postulated 된 동물 실험의 3Rs 개념 (교체, 감소, 및 개선)와 라인에1959 25.

<table border="1"><tbody><tr><td> 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 코멘트 </td></tr><tr><td> 외과 현미경 </td><td> Leica </td><td> M651 </td><td></td></tr><tr><td> 광원 </td><td> SCHOTT </td><td> KL1500LCD </td><td></td></tr><tr><td> 면봉 </td><td> NOBA Verbandmittel </td><td> 974202 </td><td></td></tr><tr><td> 압박 붕대 면봉 (5x5 cm) </td><td> Fuhrmann </td><td> 10,002 </td><td></td></tr><tr><td> povidone-요오드 솔루션 </td><td> Mundipharma </td><td> 6108022.00.01 </td><td></td></tr><tr><td> 석유 기반의 진흙 </td><td> Debika 법인 </td><td> 090148 </td><td></td></tr><tr><td> TachoSil </td><td> 다케다 제약 국제 GmbH의 </td><td> EU/1/04/277/001-004</td><td> resected 간 표면에 적용 </td></tr><tr><td> 메스 블레이드 11 호 </td><td> Pfm 의료 </td><td> 200130011 </td><td> 커프스와 stents의 작성 </td></tr><tr><td> 14 게이지 카테터 </td><td> B. 브라운 </td><td> 4268210S </td><td> PV 용 커프 </td></tr><tr><td> 18 게이지 카테터 </td><td> B. 브라운 </td><td> 4268130S </td><td> PV를 통해 재관류 </td></tr><tr><td> 24 게이지 카테터 </td><td> B. 브라운 </td><td> 4269071S </td><td> BD와 HA에 대한 스텐트 </td></tr><tr><td> 4-0 실크 봉합사 </td><td> Resorba </td><td> H3F </td><td> 간 절제술 </td></tr><tr><td> 6-0 실크 봉합사 </td><td> Resorba </td><td> H1F </td><td></td></tr><tr><td> 7-0 Prolene (폴리 프로필렌) 봉합 </td><td> Ethicon </td><td> 8701H </td><td> SHVC 및 IHVC </td></tr><tr><td> 4-0 Vicryl 봉합사 </td><td> Ethicon </td><td> V304H </td><td> 복부 폐쇄 </td></tr><tr><td> 5 ML의 주사기 </td><td> Terumo </td><td> SS + T05ES1 </td><td> 베개를 백업 </td></tr><tr><td> 생리대 </td><td> 열 </td><td> 190 X 260mm </td><td></td></tr><tr><td> 자기 fixator의 후퇴 시스템 </td><td> 정밀 과학 도구 주식회사 </td><td> 18200-01 18200-02 18200-03 18200-12 </td><td></td></tr><tr><td> 냉수 목욕 </td><td> 후버 </td><td> 740.000X </td><td> 그라프 트 보존 </td></tr><tr><td> 양극성 포셉 </td><td> Söring </td><td> MBC-200 </td><td></td></tr><tr><td> 모기 포셉 </td><td> BONIMED </td><td> 451-476-03 </td><td> 사용 두 쌍 </td></tr><tr><td> Adson 마이크로 포셉 </td><td> Dimeda </td><td> 10.176.12 </td><td></td></tr><tr><td> 구부러진 마일CRO 포셉 </td><td> AESCULAP </td><td> FD281R </td><td></td></tr><tr><td> 스트레이트 마이크로 포셉 </td><td> Bonimed </td><td> 451-476-03 </td><td></td></tr><tr><td> 구부러진 마이크로 가위 </td><td> Medicon </td><td> 05.15.83 </td><td></td></tr><tr><td> 스트레이트 마이크로 가위 </td><td> AESCULAP </td><td> FD12 </td><td> 정밀 절개 </td></tr><tr><td> 가위 </td><td> AESCULAP </td><td> BC211W </td><td></td></tr><tr><td> 마이크로 니들 홀더 </td><td> AESCULAP </td><td> FD241R </td><td> 재건 </td></tr><tr><td> 시장 - Hegar 니들 홀더 </td><td> 미즈호 Ikakogyo </td><td> 06-798-00 </td><td> 복부 폐쇄 </td></tr><tr><td> DeBakey 불독 클램프 (직선) </td><td> 울리히 </td><td> CV3054 </td><td></td></tr><tr><td> DeBakey 불독 클램프 (곡선) </td><td> CODMAN </td> &lt;TD&gt; 37-1062 </td><td></td></tr><tr><td> Satinsky 클램프 </td><td> Mizuhoika </td><td> 09-230-24 </td><td></td></tr><tr><td> 주변 혈관 클램프 </td><td> Teleflex 의료 </td><td> 353494 </td><td> 받는 사람 SHVC </td></tr><tr><td> 마이크로 혈관 클램프 (일회용) </td><td> AROSurgical 기기 주식회사 </td><td> TKM-1~60그램 </td><td> PV, 이식 IHVC, 그리고 수신자 HA </td></tr><tr><td> 마이크로 혈관 클램프 (금속) </td><td> 정밀 과학 도구 주식회사 </td><td> 18052-01 </td><td> 받는 사람 IHVC </td></tr><tr><td> Lactated 벨소리 솔루션 </td><td> Fresenius Kabi </td><td> 6150917.00.00 </td><td></td></tr><tr><td> 일반 생리 식염수 </td><td> DeltaSelect </td><td> 1299.99.99 </td><td></td></tr><tr><td> HTK 솔루션 </td><td> 프란즈 Köhler Chemie GmbH의 </td><td> 31268.00.00 </td><td> 보존 솔루션 </td></tr><tr><td> 헤파린 - 나트륨 </td><td> Ratiopharm </td><td> 5394.02.00 </td><td> 이식 재관류 전에 500 IU </td></tr><tr><td> 8.4 % 나트륨 중탄산염 </td><td> Fresenius Kabi </td><td> 4399.97.99 </td><td> reperfusion 후 0.5 ML </td></tr><tr><td> 5 % 포도당 용액 </td><td> B. 브라운 </td><td> 6714567.06.00 </td><td> reperfusion 1.0 ML </td></tr><tr><td> Cefuroxim 나트륨 </td><td> Fresenius Kabi </td><td> 38985.01.00 </td><td> 항생제, 16 밀리그램 / kg </td></tr><tr><td> Buprenorphine </td><td> 섹스 제약 </td><td> 997.00.00 </td><td> 진통제, 0.1 밀리그램 / kg </td></tr><tr><td> 집중 케어 단위 케이지 </td><td> Brinsea 제품 (주) </td><td> Vetario S10 </td><td> 수술 치료 </td></tr></tbody></table>

surgical procedures for a rat model of partial orthotopic liver transplantation with hepatic arterial reconstruction

전체 또는 일부 그라프 트를 사용하여 쥐의 Orthotopic 간 이식 (올트)는 이러한 이식을 보존하고 국소 빈혈 reperfusion - 상해 1,2, 면역 반응 3,4, 혈류 역학 5,6에 대한 연구로 이식 연구에 필수적인 실험 모델이며, 소형에 대한 크기 증후군 7. 쥐 올트는 실험 수술에서 가장 어려운 동물 모델들입니다 배울 수있는 시간이 오래 걸릴 고급 microsurgical 능력을 요구합니다. 따라서,이 모델의 사용이 제한되었습니다. 결과의 신뢰성 및 재현성 이러한 복잡한 동물 모델이 사용되는 실험의 핵심 구성 요소입니다 때문에,이 모델에 잘 표준화하고 정교한 절차에서 훈련 할 쥐 올트에 관련된 의사 필수적입니다. 첫 번째 모델은 describ 때부터 다양한 기술과 쥐의 올트의 수정은 8을보고되었지만그들은 정말 좋은 정도 재건 절차를 단순화하기 때문에 리 외하여 에드. 1973 년 9 간장 동맥 재건 10 Kamada 외하여 커프 문합 기술의 도입을 제거. 11,이 모델의 주요 발전했다 . Kamada 외.에 의해 모델에서 간 rearterialization도 제거되었습니다. 쥐가 간 이식 후 간장 동맥 흐름없이 살아남을 수 있기 때문에 간 arterialization의 가치 이상의 상당한 논란이 있었다. 그러나, arterialized 모델의 생리적 우수성이 점점 특히 담즙 덕트 시스템 8,12, 간 무결성 8,13,14를 보존의 관점에서, 인정되었습니다. 이 문서에서는, 우리는 예 생체 간 resec 후 50 % 부분 이식을 사용하여 간장 동맥 재건과 올트의 쥐 모델에 대한 자세한 수술 절차를 제시기. 과와 간장 동맥에 대한 스텐트 기술, 포털 정맥의 커프 기술, 위에와 infrahepatic 대정맥에 7-0 폴리 프로필렌 연속 봉합 : 각 혈관과 담즙 덕트의 재건 절차는 다음과 같은 방법으로 수행됩니다 담즙 덕트.

모든받는 사람 쥐 (N = 20) (각 시점에서 N = 5) 포털 reperfusion 후 (7 일) 1, 3, 24, 그리고 168 시간의 혈액 샘플링에 대한 계획 안락사까지 뚜렷한 합병증없이 살아 남았습니다. 혈액 샘플은 27 게이지 바늘로 직접 주사하여 IHVC에서 수집되었다. 10 분에 5,340 XG에서 원심 분리 후, 혈청 샘플이 획득되었으며 이식 후 간세포 손상의 정도를 반영 알라닌 아미노 전이 효소 (ALT) 수준에 대해 분석했다. 혈청 ALT 수준의 변화의 시간 코스는 그림 16에 표시됩니다. ALT 수준은 24 시간에서 피크에 도달 (± 표준 편차 의미 : 212.6 ± 67.9 IU / L)를 한 후 168 시간 (33.6 ± 6.8 IU / L)에 정상 범위 내에을 거부했다. <img alt="그림 1" src="/files/ftp_upload/4376/4376fig1.jpg" /> 그림 1. 1 일부터 포털 정맥 (PV)에 대한 커프4 게이지 카테터 및 24 게이지 카테터에서 간장 동맥 (HA) 및 담즙 덕트 (BD)를 stents. <img alt="그림 2" fo:content-width="4in" fo:src="/files/ftp_upload/4376/4376fig2highres.jpg" src="/files/ftp_upload/4376/4376fig2.jpg" /> 그림 2. 기증 쥐의 간 제거의 스키마 BD, 담즙 덕트,. HA, 간장 동맥, IHVC, infrahepatic 대정맥, PV, 포털 정맥, SHVC, suprahepatic 대정맥. <img alt="그림 3" src="/files/ftp_upload/4376/4376fig3.jpg" /> 그림 3. 기부자 작동. 가. 쥐가 자기 fixator의 후퇴 시스템을 가열 패드에 표시됩니다. 복부는 양국 확장과 중간 선 절개가 열립니다. B는. 스텐트의 삽입은 담즙 덕트에. C는. 간 재관류은 포털 정맥을 통해. 약어 설명 아르그림 2 거구나. <img alt="그림 4" src="/files/ftp_upload/4376/4376fig4.jpg" /> 4 그림. 포털 정맥에 커프의 첨부 파일. A, B. 포털 정맥 트렁크를 grasps DeBakey 불독 클램프는 금속 컵 위에 배치됩니다. 컵은 얼음 조각. C로 가득 플라스틱 상자에 장착되어 있습니다. 포털 정맥은 즉석으로 삽입됩니다. 라이. 포털 정맥의 벽이 7시에 즉석 외부 지라 정맥의 그루터기와 함께 즉석으로 everted 수 있습니다 시 위치 12시 위치에서 즉석의 확장. 전자. 포털 정맥은 즉석에서 원주 6-0 실크 스레드와 함께 고정된다. 검은 색 화살표는 지라 정맥의 그루터기를 나타냅니다. <img alt="그림 5" src="/files/ftp_upload/4376/4376fig5.jpg" /> 그림 5. 예 생체간장 동맥에 스텐트 삽입. 가. 간은 횡경막의 두 가장자리를 고정으로 고정되어 있으며, 간장 동맥은 바로 동맥에 출혈도 잡았 스레드를 잡고 당겨 있습니다. B는. 간장 동맥에 작은 절개의 앞쪽 벽이 바로 마이크로 포셉와 함께 개최됩니다 . C, D. 스텐트는 간장 동맥에 삽입하고 6-0 실크 스레드와 함께 고정된다. <img alt="그림 6" fo:content-width="3.5in" fo:src="/files/ftp_upload/4376/4376fig6highres.jpg" src="/files/ftp_upload/4376/4376fig6.jpg" /> 6 그림. 예 생체 50 % 간 절제술의 스키마는. 회색의 엽 (叶)이 제거됩니다. ACL, 앞 꼬리가있는 엽, PCL, 후부 꼬리가있는 제품 엽, LLL, 왼쪽 측면 엽, LML, 중간 엽의 왼쪽 부분, 중간 엽의 RML, 오른쪽 부분, SRL, 우수한 오른쪽 측면 엽, IRL, 열악한 오른쪽 측면 엽. 그림 7. 전 생체 50 % 간 절제술. 중간 엽. C의 왼쪽 부분의 작은 꽃자루의. 뒤쪽 꼬리가있는 엽의의 작은 꽃자루의 결합. 나. 결합. 50 % 절제 전에 간. 라. 50 % 절제 후 간. <img alt="그림 8" src="/files/ftp_upload/4376/4376fig8.jpg" /> 그림 8. suprahepatic 대정맥의 예 생체 plasty. 가. 간은 모기 포셉과 격막의 양쪽 가장자리를 고정으로 고정됩니다. 나. 유지 봉합을 7-0 폴리 프로필렌으로이 두 코너에 부착된다. <img alt="그림 9" fo:content-width="4in" fo:src="/files/ftp_upload/4376/4376fig9highres.jpg" srC = &quot;/ files/ftp_upload/4376/4376fig9.jpg&quot;/&gt; 9 그림. 받는 사람 쥐에 이식 주입의 스키마. 재건 절차는 7-0 연속 봉합, 커프 기술에 의해 포털 정맥 (PV), 그리고 간장과 위에와 infrahepatic 대정맥 (SHVC 및 IHVC)에 대해 수행됩니다 스텐트 기술에 의해 동맥 (HA) 및 담즙 덕트 (BD). <img alt="그림 10" src="/files/ftp_upload/4376/4376fig10.jpg" /> 10 그림. 기본 간을 제거 할 때까지받는 작업. 가. 복부은 중간 선 절개가 열립니다. 나. 오른쪽 부신 정맥은 출혈도 잡았 있습니다. C는. 원시 간은 excised 수 있습니다. 약어는 그림 2에 설명되어 있습니다. <img alt="그림 11" src="/files/ftp_upload/4376/4376fig11.jpg" /> 11 그림. suprahe의 문합patic 대정맥. 는 나. suprahepatic 대정맥의 주변 혈관 클램프는 석유 기반 진흙의 덩어리에서 해결되었습니다. 두 코너의 숙박 봉합은 superiolaterally 문합 부위를 확대하기 위해 부드러운 미끄러지는 유지됩니다. C는. 진행 뒤 행의 연속 intraluminal 봉합사가. D는. 진행 앞 행의 연속 봉합은. <img alt="그림 12" src="/files/ftp_upload/4376/4376fig12.jpg" /> 그림 12. 포털 정맥의 재건. A, B. 포털 정맥을 클램핑 모기 포셉은 석유 기반의 진흙에서 수정 및 간 hilum쪽으로 당겨 있습니다. CF는. 포털 정맥에 즉석의 삽입. <img alt="그림 13" src="/files/ftp_upload/4376/4376fig13.jpg" /> 13 그림. 간장 동맥의 재건. A, B. </stron<html> g 적절한 간장 동맥 (PHA)과 gastroduodenal 동맥 (GDA)의 분기에서받는 일반적인 간장 동맥 (CHA)로 스텐트의&gt; 삽입. <img alt="그림 14" src="/files/ftp_upload/4376/4376fig14.jpg" /> 14 그림. infrahepatic 대정맥의 문합. infrahepatic 대정맥의. 두 모서리에있는 숙박 봉합. 나. 뒤쪽 행의 연속 봉합 해. 다. 앞 행의 연속 봉합 해. 라. Reperfusion. 약어는 그림 2에 설명되어 있습니다. <img alt="그림 15" src="/files/ftp_upload/4376/4376fig15.jpg" /> 그림 15. 모든 재건 절차가 완료됩니다. 약어는 그림 2에 설명되어 있습니다. <img alt="&lt;/ HTML">&quot;그림 16&quot;SRC = &quot;/ files/ftp_upload/4376/4376fig16.jpg&quot;/&gt; 그림 16 혈청 알라닌 아미노 전이 효소 (ALT) 수준의 변화의 시간 코스 수술 (N = 20, N 각 시점에서 = 5).. 데이터는 표준 편차를 나타내는 오류 바가있는 수단으로 표현됩니다. ALT 수준은 24 시간에서 피크를 (212.6 ± 67.9 IU / L)에 도달 한 후 168 시간 (33.6 ± 6.8 IU / L)에 정상 범위 내에을 거부했다.

Watch this Scientific Journal Video about Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction at JoVE.com

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

쥐의 Orthotopic 간 이식은 생명 의학 연구에 필수적인 실험 모델입니다. 여기 50 % 부분 이식을 사용하여 간장 동맥 재건과 orthotopic 쥐의 간 이식에 대한 수술 절차를 제시한다.

간 간선 재건과 부분 Orthotopic 간 이식의 쥐 모델에 대한 외과 절차

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

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36.0K 조회수.  RWTH-Aachen University. 쥐의 Orthotopic 간 이식은 생명 의학 연구에 필수적인 실험 모델입니다. 여기 50 % 부분 이식을 사용하여 간장 동맥 재건과 orthotopic 쥐의 간 이식에 대한 수술 절차를 제시한다.

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

Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant vasculopathy (TVP).
The commonly used animal model for cardiovascular chronic rejection studies is the heterotopic heart transplant model performed in laboratory rodents. This model is used widely in experiments since Ono and Lindsey (3) published their technique. To analyze the findings in the blood vessels, the heart has to be sectioned and all vessels have to be measured.
Another method to investigate chronic rejection in cardiovascular questionings is the aortic transplant model (1, 2). In the orthotopic aortic transplant model, the aorta can easily be histologically evaluated (2). The PVG-to-ACI model is especially useful for CAV studies, since acute vascular rejection is not a major confounding factor and Cyclosporin A (CsA) treatment does not prevent the development of CAV, similar to what we find in the clinical setting (4). A7-day period of CsA is required in this model to prevent acute rejection and to achieve long-term survival with the development of TVP.
This model can also be used to investigate acute cellular rejection and media necrosis in xenogeneic models (5).

This video demonstrates the orthotopic aortic transplant model as a simple model to study the development of transplant vasculopathy (TVP) in rats.

Orthotopic 대동맥 이식 : 만성 Vasculopathy의 개발 연구에 생쥐 모델

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant ...

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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.

쥐에게 Orthotopic 간 이식

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

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.

이소성 보조 쥐 간 이식 급성 간 장애에 포털 정맥 Arterialization를 유량 조절

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

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.

쥐 간 문부 클램프 모델을 사용하여 직접 분절 인트라 간 납품 방법

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

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.

다시는 vivo에서 선박 지향 70% 쥐 부분 간 이식 Arterialized Hepatectomy

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

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.

Orthotopic의 쥐 모델 간 이식 Suprahepatic 베 나 정 맥 재건에 대 한 새로운 자석 문 합 기술을 사용 하 여

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

Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of renal replacement therapy. Over the past few decades, the rat kidney transplantation model has been used to study the immunological phenomena of rejection and tolerance. This model has become an indispensable tool to test new immunomodulatory pharmaceuticals and regimens prior to proceeding with expensive preclinical large animal studies.
This protocol provides a detailed overview of how to reliably perform orthotopic kidney transplantation in rats. This protocol includes three distinctive steps that increase the probability of success: perfusion of the donor kidney by flushing through the portal vein and the use of a cuff system to anastomose the renal veins and ureters, thereby decreasing cold and warm ischemia times. Using this technique, we have achieved survival rates beyond 6 months with normal serum creatinine in animals with syngeneic or tolerant kidney transplants. Depending on the aim of the study, this model can be modified by pre- or posttransplant treatments to study the acute, chronic, cellular, or antibody-mediated rejection. It is a reproducible, reliable, and cost-effective animal model to study different aspects of kidney transplantation.

The purpose of this manuscript and protocol is to explain and demonstrate in detail the surgical procedure of orthotopic kidney transplantation in rats. This method is simplified to achieve the correct perfusion of the donor kidney and shorten the reperfusion time by using the venous and ureteral cuff anastomosis technique.

직교 쥐 신장 이식: 새롭고 단순화 된 외과 접근

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of ...

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% 부분 간 절제술과 아세트아미노펜을 결합하여 급성 간 기능 부전의 쥐 모델 생성

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.

직교 간 이식의 쥐 모형에 있는 동맥 재연결 후에 감소된 합병증

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

Left Lung Orthotopic Transplantation in a Juvenile Porcine Model for ESLP

Lung transplantation is the gold-standard treatment for end-stage lung disease, with over 4,600 lung transplantations performed worldwide annually. However, lung transplantation is limited by a shortage of available donor organs. As such, there is high waitlist mortality. Ex situ lung perfusion (ESLP) has increased donor lung utilization rates in some centers by 15%-20%. ESLP has been applied as a method to assess and recondition marginal donor lungs and has demonstrated acceptable short- and long-term outcomes following transplantation of extended criteria donor (ECD) lungs. Large animal (in vivo) transplantation models are required to validate ongoing in vitro research findings. Anatomic and physiologic differences between humans and pigs pose significant technical and anesthetic challenges. An easily reproducible transplant model would permit the&#160;in vivo&#160;validation of current ESLP strategies and the preclinical evaluation of various interventions designed to improve donor lung function. This protocol describes a porcine model of orthotopic left lung allotransplantation. This includes anesthetic and surgical techniques, a customized surgical checklist, troubleshooting, modifications, and the benefits and limitations of the approach.

This protocol describes a juvenile porcine model of orthotopic left lung allotransplantation designed for use with ESLP research. Focus is made on anesthetic and surgical techniques, as well as critical steps and troubleshooting.

ESLP를 위한 어린 돼지 모델에서의 좌측 폐 정형외과적 이식

Lung transplantation is the gold-standard treatment for end-stage lung disease, with over 4,600 lung transplantations performed worldwide annually. ...