마크 제이 컨즈는 (의사 및 캐나다의 왕립 외과 대학을 통해 수여) UBC 임상 탐정 프로그램 및 UBC 4YF 박사 원정대의 지원을받습니다. 존 H. 보이드는 건강 연구 학술위한 국가 Sanitorium 협회와 마이클 스미스 재단이다. CIHR을 통해 자금. 저자는 설치 및 관류 장비의 유지 보수와 그들의 도움 박사 엠 알라 리처드 Wamboldt을 감사드립니다.

모든 동물은 보관과 전국 및 관리 및 실험 동물의 사용을위한 제도적 지침에 따라 마음에 든다고했다. 이 프로토콜에 대한 윤리 승인은 브리티시 컬럼비아의 동물 관리위원회의 대학에 의해 수여되었다. 300 사이 계량 남성, 흰쥐 - 450 g을이 프로토콜에 사용 하였다. 1. 기부자 운영 <ol><li> 3 방향 스톱 콕 장거리 정맥 내 (IV) 카테터 튜브에 연결된 원통형 플라스크에 심정지 (RT)의 약 100 mL의 유무. 중력에 의해 심정지 전달을 가능하게 약 80cm 수술 표면 위의 플라스크를 상승 스탠드를 사용합니다. </li><li> 적절하게 구조를 시각화하기 위해, 수술 돋보기 한 쌍 또는 해부 현미경 중 하나를 사용하십시오. 참고 : 우리는 현재 3.4 쌍안경 운영 현미경을 사용 - 21.3X 배율. </li><li> 마취 실에서 기증자를 놓고 4 마취 유도 - 5 %의 이소 플루 란을. </lI&gt; <li> 운영 플랫폼에 쥐를 전송하고 1 코 콘에 의해 마취 유지 - 2 % 이소 플루 란을. 건조를 방지하기 위해 동물의 눈에 수의사 연고를 적용합니다. 25 G 바늘과 미다 졸람 (2 ㎎ / ㎏) 내 복강을 관리합니다. </li><li> 수술 가위를 사용하여, 하악에 조류 늑골의 검상 돌기에서 기증자를 면도. (이후 IV 액세스) 왼쪽 사타구니의 작은 영역을 면도. 수술 표면에 제모 제를 적용, 약 5 분 정도 기다린 거즈 조각으로 머리를 제거합니다. </li><li> 포비돈 - 요오드 또는 chlorexidine 기반 솔루션과 수술 사이트를 (우리는 단지 클로르헥시딘 사용) 준비. 0.5 % 리도카인 피하 - 0.1와 절개 사이트를 침투. </li><li> 마취 깊이와 생체 신호의 모니터링을위한 무료 오른쪽 뒷다리를 떠나, 접착 테이프와 운영 플랫폼에 앞발과 왼쪽 뒷다리를 고정합니다. </li><li> 페달 핀치에 의해 적절한 마취 깊이를 확인한 후 사용 왼쪽 사타구니 주름에 절개를22 블레이드 메스. 무딘 절개하여 좌측 대퇴 정맥을 노출하고 다음과 같이 IV 액세스를 얻을 : <ol><li> 부드럽게 대퇴 정맥을 덮는 조직을 철회하고, 24 G 4과 정맥을 cannulate. 식염수 채워진 IV 튜브의 짧은 길이로 IV를 연결하고 접착 테이프로 장소에 튜브를 고정합니다. 참고 : 쥐에 대퇴 정맥 삽관에 대한 절차는 제스 퍼슨 및 동료 (14)에 의해 다른 곳에서 설명되어 있습니다. </li><li> IV의 정확한 위치를 확인하기 위해 혈액을 대기음 부드럽게 IV 튜브에 식염수로 채워진 10 ML의 주사기를 연결합니다. </li><li> IV 튜브를 통해 미분 획 헤파린 500 IU, 이후 3 튜브를 세척 - - (300)를 주입 식염수 5 ㎖. </li></ol></li><li> 다음 다음, 도너를 tracheotomize : <ol><li> 22 블레이드 메스를 사용하여 경정맥 노치과 하악골 사이의 부드러운 조직에서 중간 선 절개를합니다. 멧젠를 사용하여 중간 선에서 갑상선의 캡슐을 침투무딘 절개를 사용 바움 가위, 별도의 로브. </li><li> 무딘 절개를 사용하여 기관의 전방 표면을 노출 정중선에 목의 근육 스트랩을 분리한다. </li><li> 퉁명스럽게 기관 주위에 원주면을 해부 곡선 집게를 사용합니다. 4-0 실크 넥타이 기관을 둘러싸. </li><li> 홍채 가위를 사용하여, 갑상선 연골에 약 5mm 열등, 전방 기관에 횡 절개를합니다. 조심스럽게 기관 캐뉼라 (14 G IV)를 소개하고 4-0 실크 넥타이를 사용하여 제자리에 고정합니다. </li><li> 인공 호흡기에 기관 정맥을 연결합니다. 인공 호흡기 회로를 통해 산소와 이소 플루 란의 흐름을 리디렉션 및 속도와 무게 (13)에 의해 예측 호흡량에서 기증자를 환기 시키십시오. </li></ol></li><li> xipisternum 아래에 경정맥 노치의 절개를 연장 (22 블레이드 메스를 사용하여) 가슴의 중간 선에 절개를합니다. </li><li> 중간 선에 머물고, 뼈 커터를 사용하여 중간 흉골 절개를 수행합니다. 자기 유지 권취으로 흉골의 가장자리 후퇴. 심낭 및 흉막 충치를 입력합니다. </li><li> 흉선을 수행합니다. 먼저 퉁명스럽게 중간 선에서 흉선을 분할하고 무딘과 날카로운 해부의 조합을 사용하여 주변의 구조에서 분리하는 가장 쉬운 방법입니다. 참고 : 흉선 멀리 우수한 흉골 가장자리에서 해부시 내흉 동맥의 기원은 부상의 원인이 될 수 있습니다. 출혈을 방지하기 위해, 지혈 클립이 지점에서 흉선을 분리하기 전에 적용될 수있다. </li><li> Metzenbaum 가위를 사용 및 / 또는 날카로운 로어, 하대 정맥 (IVC)에서 멀리 지방 근교 대정맥 해부. IVC는 하방 다이어프램에, 우량 CAVO - 심방 접합에서 지방과 결합 조직의 상대적 없는지 확인합니다. </li><li> 날카로운 로어, 원주없는 상대 정맥 (SVC)를 사용하고 4-0 실크 넥타이를 둘러싸. </li><li> 다음에, 해부주변 구조에서 무료로 대정맥을 떠났고, 근위 및 원위 4-0 실크 넥타이와 그것을 결찰. 왼쪽 쇄골 하 동맥을 노출 맥의 중간 부분을 절제. </li><li> 원주 무료 날카로운 로어를 사용하여 대동맥 궁 혈관. 무명 동맥 근위부와 원위부 수술 클립을 적용하고, 클립 사이를 나눕니다. 왼쪽 경동맥과 좌측 쇄골 하 동맥 않은 클립을 남겨주세요. </li><li> 다음, 24 G IV 카테터 SVC를 cannulate. 이전에 배치 4-0 실크 넥타이와 카테터를 고정합니다. </li><li> 편법,하지만 부드러운 이식 수확을 준비합니다. </li><li> 날카로운 가위를 사용하여, 횡격막 표면에 IVC 그냥 우수한 나눕니다. 근처의기도 구조가 손상되지 않도록 보장의 삽관 사이트에 SVC 우수한 나눈다. </li><li> 인공 호흡기의 전원을 끄고 기관 정맥을 분리합니다. 근위 기관을 가로로 쪼개다. </li><li> 집게로 횡단 기관을 잡고 마음을 제거폐 KO-블록. 심장 - 폐 이식이 제거 될 때이 날카로운 해부와 부드러운 견인을 필요로 할 것이다. 기본 식도에서 기관을 분리합니다. 그것의 긴 부분은 수확 후 그대로 유지되도록, 하행 대동맥 부상을 피한다. </li></ol> 심장 - 폐 이식 2. 준비 참고 : 프로 시저의이 부분을 완료하는 동안, 조수는받는 사람 쥐를 마취 이식 주입을 준비해야한다. <ol><li> 아래로 향하게 마음의 전방 표면과 수술 거즈에 심장 - 폐 이식을 놓습니다. 날카로운 해부를 사용하여 후방기도에 손상을 방지 남아있는 식도를 제거합니다. </li><li> 하행 흉부 대동맥을 찾아 16 G의 무딘 팁 캐뉼라를 삽입합니다. 4-0 실크 넥타이와 장소에 캐 뉼러를 고정합니다. </li><li> 대동맥 궁 혈관을 찾아 왼쪽 쇄골 하 동맥에 외과 용 클립을 적용합니다. 취소 잘린 왼쪽 경동맥을 남겨 서브에 대한필연 드 - 방송. </li><li> 16 G 대동맥 캐뉼라로 심정지 튜브를 접속함으로써 심정지의 제 투여를 관리. 경동맥은 일시적으로 심정지의 적절한 공급을 보장하기 위해 집게로 폐색해야합니다. 주 : 부착 대동맥 캐뉼라를 갖는 이점에 원하는대로 심정지 연속적 및 / 또는 간헐적으로 투여 될 수 있다는 점이다. 45 초 - 30 5 ml의 환약 15 분 - 간헐적 투여를 들어, 우리는 일반적으로 심정지마다 10을 제공합니다. </li><li> Metzenbaum 가위를 사용 및 / 또는 날카로운 로어, 멀리 대동맥에서 요정 대동맥의 지방을 해부하다. 대동맥은 삽관 사이트에 왼쪽 쇄골 하 동맥 (말초 아치)에서 지방과 결합 조직의 상대적으로 자유로운 지 확인합니다. </li><li> 다음, 날카로운 로어를 사용하여 왼쪽 mainstem 기관지를 노출하고 4-0 실크 넥타이와 근위을 결찰. 홍채 가위를 사용하여 합자 왼쪽 기관지의 말단을 가로로 쪼개다. 왼쪽 폐를 결찰에 의해 왼쪽 폐 절제술을 수행4-0 실크 넥타이와 동맥과 정맥. 합자에 말단 횡단면과 왼쪽 폐를 제거합니다. </li><li> 상기와 같이, 오른쪽 상단 및 중간 엽을 제외하고 다른 모든 폐 엽을 제거합니다. 명중 폐엽을 수행하는 과정에서 부상 IVC 피한다. </li><li> 대동맥 정맥에 심정지 튜브를 연결하고받는 사람이 준비되는 동안 지속적으로 심정지를 관리 할 수​​ 있습니다. 멸균 용기 (예를 들어, 비커)에 심장 - 폐 이식을 놓습니다. </li></ol> 3.받는 사람 운영 <ol><li> 상기 도너 조작에 따라 수신자 마취. 건조에서 동물의 눈을 보호하기 위해 수의사 연고를 사용합니다. 페달 핀치에 의해 자주 마취를 확인합니다. </li><li> 활력 징후 및 마취 깊이를 모니터 무료 오른쪽 앞다리를 떠나, 기증자 작업에 따라이 시간을 동물을 배치합니다. </li><li> 조류 늑골의 검상 돌기에서 음경에 복부를 면도. (이후 IV 액세스) 왼쪽 사타구니의 작은 영역을 면도. 적용수술 표면에 에이전트를 제모, 약 5 분 정도 기다린 거즈 조각으로 머리를 제거합니다. </li><li> 포비돈 - 요오드 또는 chlorexidine 기반 솔루션과 수술 사이트를 준비. 0.5 % 리도카인 피하 - 0.1와 절개 사이트를 침투. </li><li> Tracheotomize 위의 지시에 따라 수신자를 환기 시키십시오. 또한, 코 콘 마취하에받는 사람을 유지한다. </li><li> 전술 한 바와 같이 대퇴 정맥에 카테터를 소개한다. IV 튜브를 통해 미분 획 헤파린 500 IU, 이후 3 튜브를 세척 - - (300)를 주입 식염수 5 ㎖. </li><li> 음경에 조류 늑골의 검상 돌기에서 22 블레이드 메스로 중간 선 복부 절개를하여 개복술을 수행합니다. 자체 유지 견인기를 사용하여 복벽을 후퇴. 다음으로, 우량와 쥐의 왼쪽에 창자를 철회. 따뜻한 식염수를 적신 거즈를 감싸. </li><li> 크게 상부 재를 통해 해부하여 IVC 및 복부 대동맥을 노출TRO-복막 지방. </li><li> 곡선 용기 클램프가 가능하고 준비합니다. </li><li> 원주 근위 및 원위 하대 정맥과 대동맥를 해제하고 4-0 실크 넥타이로 둘러싸. 이 사이트 사이의 공간의 3 CM - 약 2가 있어야합니다. </li><li> 조심스럽게 하대 정맥과 대동맥 모두의 충분한 부분이 클램프의 턱 위에 노출되는 것을 보장, 곡선 용기 클램프를 적용합니다. </li><li> 식염수로 채워진 1 ML의 주사기에 연결된 25 G 바늘 IVC의 앞쪽 벽에 절개를합니다. 기증자 IVC 구멍의 길이에 맞게 포츠 가위로 절개를 확장 할 수 있습니다. </li><li> 컨테이너에서 심장 - 폐 이식을 제거하고는 심정지에서 분리합니다. 정맥 문합을 수행 할 최적의 위치에받는 사람의 복부에 이식을 놓습니다. 참고 : 심장 - 폐 이식이 궁극적으로 복부의 방향으로 정점을 가리키는, 약간 비스듬하게 지향한다 낮은 사분면을 떠났습니다. </li><li> 발 뒤꿈치를 고정발가락은 9-0 나일론 봉합사와 봉합의 끝납니다. 봉합의 긴 팔에 부착 된 바늘, 이후​​에 연결되는 봉합사의 짧은 끝을 떠나, 각 끝 부분에 안전한 매듭을 묶어. 심정지의 복용량을 관리합니다. </li><li> 패션을 실행에, 반대 짧은 봉합 팔에 봉합 라인과 넥타이의 완전한 절반. 심정지의 복용량을 관리합니다. </li><li> 봉합 라인의 나머지 절반을 완료하고 넥타이. 정맥 문합이 완료됩니다. 심정지의 복용량을 관리합니다 </li><li> 동양은 복부의의를 향한 마음의 정점을 가리키는 심장 - 폐 이식은 하복부를 떠났다. 정맥 문합이 꼬거나 비틀어되지 않았는지 확인합니다. </li><li> 긴장 - 무료 및 취소 꼬여 구성에서 IVC와 대동맥 문합을 모두 유지,받는 사람 대동맥에 도달해야합니다 기증자의 대동맥의 길이를 평가합니다. </li><li> 홍채 가위 (왼쪽 쇄골 하 동맥 원위부) 하행 대동맥을 나눈다. </li><li> incisio 확인N 식염수로 채워진 1 ML의 주사기에 연결된 25 G 바늘받는 사람 대동맥의 앞쪽 벽에. 도너 대동맥 오리피스의 길이와 일치하도록 포츠 가위로 절개 확장. </li><li> 9-0 나일론 봉합사를 이용하여 대동맥받는 사람에게 기증자의 대동맥의 발가락 (우수한 측면을) 앵커. 이어서, 대동맥의 중간 형태에 바늘을 통과 패션에서 실행 문합의 절반을 완성. </li><li> 대동맥 문합 뒤꿈치에서 봉합 라인 (스티치의 방향 전환)의 방향을 반대로하고, 패션에서 실행 문합의 좌우 절반을 완료. </li><li> 천천히 SVC의 IV 카테터를 제거하고 수술 클립 SVC를 결찰. </li><li> 드 공기 않은 잘린 왼쪽 경동맥의 위치에 의해 대동맥. 공기가 자유롭게 대피 할 수 있도록 수직 위치에서 열기 동맥을 잡으십시오. </li><li> 간단히 D 곡선 용기 클램프의 턱을 열고 (경동맥은 피 할 수 있도록전자 공기) 2 - 3 초. 선박 클램프를 다시 적용 할 수 있습니다. </li><li> 경동맥을 외과 용 클립을 적용합니다. 곡선 용기 클램프를 제거합니다. </li><li> 봉합 라인에서 출혈이 있는지 확인합니다. 존재하는 경우, 9-0 나일론 봉합사 (출혈의 정도에 따라 다름)의 길이와 수술 거즈 또는 수리와 부드러운 압축을 적용합니다. </li><li> 기증자 심장 분 이내에 치고 다시 시작해야합니다. </li><li> 받는 사람의 생체 신호, 마취의 깊이에주의하고 필요에 따라 볼륨 소생술을 관리 할 수​​ 있습니다. </li><li> 중, 실험 엔드 포인트에 따라 동물의 복부를 닫고 마취에서 회복하자, 또는 심장 혈관 평가를 위해 동물을 준비합니다. 참고 : 이식 평가의 예는 부하에 따라 부하 독립적 인 혈역학의 생체 측정, 시험 관내 랑겐 돌프 함수의 측정 및 작업 심장 모드, (생존 수술) 심 초음파 나 MRI 조사에 포함. </li><li> 에서급성 터미널 실험의 끝, 동물 방혈에 의해 안락사된다. </li></ol>

<ol>
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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=The+effect+of+load+on+atrophy,+myosin+isoform+shifts+and+contractile+function:+studies+in+a+novel+rat+heart+transplant+preparation.">The effect of load on atrophy, myosin isoform shifts and contractile function: studies in a novel rat heart transplant preparation.</a> J Mol Cell Cardiol. 27 (1), 407-417 (1995).</li><li>Yokoyama, H., Ohmi, M., Murata, S., Nakame, T., Tabayashi, K., Mohri, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proposal+of+a+working+left+heart+model+with+a+heterotopic+transplantation+technique+in+rats.">Proposal of a working left heart model with a heterotopic transplantation technique in rats.</a> J Heart Lung Transplant. 14 (4), 706-712 (1995).</li><li>Maruyama, T., Swartz, M. T., McBride, L. R., Pennington, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Working+heart+model+of+heterotopic+heart-lung+transplantation+in+rats.">Working heart model of heterotopic heart-lung transplantation in rats.</a> J Thorac Cardiovasc Surg. 107 (1), 210-215 (1994).</li><li>Pacher, P., Nagayama, T., Mukhopadhyay, P., B&#225;tkai, S., Kass, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+cardiac+function+using+pressure-volume+conductance+catheter+technique+in+mice+and+rats.">Measurement of cardiac function using pressure-volume conductance catheter technique in mice and rats.</a> Nat Protoc. 3 (9), 1422-1434 (2008).</li><li>Jespersen, B., Knupp, L., Northcott, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Femoral+arterial+and+venous+catheterization+for+blood+sampling,+drug+administration+and+conscious+blood+pressure+and+heart+rate+measurements.">Femoral arterial and venous catheterization for blood sampling, drug administration and conscious blood pressure and heart rate measurements.</a> J Vis Exp. (59), 3496-3410 (2012).</li><li>Habertheuer, A., Kocher, 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=Innovative,+simplified+orthotopic+lung+transplantation+in+rats).">Innovative, simplified orthotopic lung transplantation in rats).</a> J Surg Res. 185 (1), 419-425 (2013).</li></ol>

저자가 공개하는 게 없다.

여기에 설명 된 기술과 성공은 여러 가지 요인에 입각한다. 그 중 주요 균일 이식 냉각 완전한 심정지 체포, 총 허혈 시간을 최소화을 보장, 안전하고 최소한의 혈액 손실과 관련된 세심한 수술 기법을 채택, 모두 기증자와받는 사람 동물의 안정성을 보장하고, 적절하게 탈 방송 이식을 할 것이다. 위의 인정으로, 기술의 기술적 인 복잡성은 최고 제한 사항입니다. 우리는 여?...

The first rodent model of heterotopic abdominal heart transplantation (HAHT) was described by Abbott and colleagues in 19641. This technique, and subsequent modifications have been widely applied to characterize transplant graft function and immunologic status. The majority of HAHT techniques described involve a non-volume loaded heart2,3. Models of HAHT involving volume-loaded ventricles have been described, but they are frequently limited in one or more respects.
Heterotopic abdominal heart-lung transplantation (HAHLT) with a volume-loaded left ventricle (LV) has been described previously. Chen and colleagues4, and subsequently Ibrahim and colleagues5 described HAHLT with a single aorto-aortic (donor ascending to recipient abdominal aorta) anastomosis. The only volume load presented to the ventricle in this circulation is the coronary venous return. Asfour and colleagues described a HAHT technique in which the lung circuit was eliminated by anastomosing donor pulmonary artery (PA) to donor left atrium (LA)6. In this circulation, venous inflow to right ventricle (RV) occurs via a donor SVC to recipient IVC anastomosis, and the subsequent LV load is ejected into the aorto-aortic anastomosis. Cardiac function was partially assessed in vivo, and also in vitro using a Langendorff rig. Figueiredo and colleagues described a HAHLT model similar to our own7, but in mice. Venous inflow to the RV occurs via donor SVC to recipient IVC anastomosis. Blood subsequently passes through the single lung circulation and LV load is ejected into the aorto-aortic anastomosis. Cardiac function in their study was assessed by magnetic resonance imaging (MRI). Wen and colleagues described a unique HAHT technique in which the LV is loaded by means of a recipient aorta to donor LA anastomosis8. The LV, therefore, fills at systemic pressures. Cardiac function, and whether LV stroke volume is ejected antegradely in their model was not assessed.
Many of the techniques referenced above involve non-physiologic LV loading conditions, including the techniques whose partial LV load is represented only by coronary venous return. On the other hand, many techniques do approach physiologic LV loading. The majority of these techniques, as with the technique of Asfour and colleagues, omit the pulmonary circulation and utilize a donor PA to donor LA anastomosis6,9. The circulation described by Galinanes and colleagues10 employs a direct recipient cava to donor LA anastomosis, omitting the pulmonary circulation and the right heart. Yokoyama and colleagues achieve the same effect by ligating the donor PA and creating an interatrial communication in the donor heart (omitting donor lung and right heart circulations)11. The circulation of Maruyama and colleagues12 involves an anastomosis between donor left PA and recipient Ao, which permits LV filling via the pulmonary circulation as a conduit, but effectively excludes the right heart.
In cases where near physiologic loading conditions were met, we advance the technique of HAHLT in 2 major respects. First, to our knowledge, the exact configuration we report has not been described in rats. It is possibly the most versatile circulation for investigators wishing to study the physiology, structure, and immunology of the transplanted heart-lung graft. Second, we describe how the function of the transplant graft can be directly characterized in vivo. For this application, pressure-volume conductance catheters can be introduced directly into the LV apex of the transplant graft, which allows for complete cardiac functional characterization. 
The technique described here can be applied to both acute and chronic studies of transplant graft function, while the functional assessment may be performed either in vivo or in vitro. We present a model in which the loading conditions can be near physiologic, however the degree of ventricular loading may be manipulated both acutely and chronically by diverting venous return towards or away from the graft. Afterload conditions can also be manipulated. Because the lung and its airway are retained in this transplant configuration, investigators can re-ventilate the donor lung acutely. Uniquely, lung re-ventilation changes the composition of blood perfusing the transplant coronary arteries. Under non-ventilated conditions, blood ejected from the donor aorta is deoxygenated, and mixes with oxygenated blood in the recipient aorta. Under acutely ventilated conditions, ejected blood becomes oxygenated. Thus, transplant graft function can be compared under ventilated and non-ventilated conditions, and also under variably loaded conditions.
The protocol below describes important modifications to previously described HAHLT donor and recipient operations. It also describes an optimal technique for protecting the transplant graft throughout the period of ischemia (time between donor explant and recipient implant). Advantages of this technique include physiologic conditions potentially approaching that of an orthotopically transplanted graft, and a wide range of investigative applications. An important limitation is its technical complexity. With adequate mentoring and practice, the advantages of this technique will likely outweigh the challenges in adopting it.

<table><tbody><tr><td>Celsior Cardioplegic Soution</td><td>Genzyme</td><td></td><td>The solution is kept on ice throughout the procedure. We prepare our own solution, with slight modifications.</td></tr><tr><td>Rodent Ventilator</td><td>Harvard Apparatus</td><td>Model 683</td><td></td></tr><tr><td>Vital Sign Monitor</td><td>Nonin</td><td>Model 9847V</td><td>Displays SpO&lt;sub&gt;2&lt;/sub&gt; and heart rate.</td></tr><tr><td>IV Cannulae</td><td>Jelco</td><td>3063</td><td>24 - 26 G x 3/4&quot; cannulae.</td></tr><tr><td>IV Tubing</td><td>CareFusion</td><td>MP9259-C</td><td>Short-length connector tubing (18 cm).</td></tr><tr><td>Surgical Clips</td><td>Teleflex Medical</td><td>001204</td><td>Horizon titanium ligating clips.</td></tr><tr><td>Sutures</td><td>Ethicon, Sharpoint</td><td>LA54G, AK-0107</td><td>3-0 silk reel, and 9-0 prolene suture (single-armed, DR5 needle).</td></tr><tr><td>Surgical Instruments</td><td>Not Applicable</td><td>Not Applicable</td><td>The instruments used are generic, and can be purchased from any surgical supply company.</td></tr></tbody></table>

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.

위에서 설명한 HAHLT 기술은 고도의 기술과 세부 사항입니다. 표에 세심한주의를 필요로 한 하이라이트 실패 절차 대 성공과 관련된 중요한 요소의 일부, 그리고 기술적 인 문제를 해결하기위한 가이드로 사용할 수 있습니다. 받는 사람 대동맥이 클램핑 해제 한 후, 이식 관상 동맥은 산소가 혈액으로 채우기 위해 볼 수 있어야합니다. 따라서, 심근는 ...

Watch this Scientific Journal Video about Rat Heterotopic Abdominal Heart/Single-lung Transplantation in a Volume-loaded Configuration at JoVE.com

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

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.

쥐 이소성 복부 심장 / 볼륨로드 구성에서 단일 폐 이식

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

rat-heterotopic-abdominal-heartsingle-lung-transplantation-volume

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

8.3K 조회수.  UBC James Hogg Research Center, St. Paul's Hospital. 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.

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

Heterotopic Heart Transplantation in Mice

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable for studying rejection and immune response now that newer transgenic and gene knockout mice are available, and a large number of immunologic reagents have been developed. The heart transplant model is less stringent than the skin transplant models, although technically more challenging. We have developed a modified technique and have completed over 1000 successful cases of heterotopic heart transplantation in mice. When making anastomosis of the ascending aorta and abdominal aorta, two stay sutures are placed at the proximal and distal apexes of recipient abdominal aorta with the donor s ascending aorta, then using 11-0 suture for anastomosis on both side of aorta with continuing sutures. The stay sutures make the anastomosis easier and 11-0 is an ideal suture size to avoid bleeding and thrombosis.
When making anastomosis of pulmonary artery and inferior vena cava, two stay sutures are made at the proximal apex and distal apex of the recipient s inferior vena cava with the donor s pulmonary artery. The left wall of the inferior vena cava and donor s pulmonary artery is closed with continuing sutures in the inside of the inferior vena cava after, one knot with the proximal apex stay suture the right wall of the inferior vena cava and the donor s pulmonary artery are closed with continuing sutures outside the inferior vena cave with 10-0 sutures. This method is easier to perform because anastomosis is made just on the one side of the inferior vena cava and 10-0 sutures is the right size to avoid bleeding and thrombosis. In this article, we provide details of the technique to supplement the video.

The mouse heterotopic heart transplantation model has been proven by many investigators to be an important method for studying mechanisms of rejection and immune response. However, the techniques involved are still challenging. By modifying standard techniques we have had success with more than 1000 transplants.

마우스의 Heterotopic 심장 이식

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable ...

연구

생물학

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 ...

의학

Murine Heterotopic Heart Transplant Technique

It is now over forty years since this technique was first reported by Corry, Wynn and Russell. Although it took some years for other labs to become proficient in and utilize this technique, it is now widely used by many laboratories around the world. A significant refinement to the original technique was developed and reported in 2001 by Niimi. Described here are the techniques that have evolved over more than a decade in the hands of three surgeons (Plenter, Grazia, Pietra) in our center. These techniques are now being passed on to a younger generation of surgeons and researchers.
Based largely on the Niimi experience, the procedures used have evolved in the fine details - details which we will endeavor to relate here in such a way that others may be able to use this very useful model. Like Niimi, we have found that a video aid to learning is a priceless resource for the beginner.

The manuscript describes the steps required to perform the heterotopic heart transplant in the mouse.

쥐 이소성 심장 이식 기술

It is now over forty years since this technique was first reported by Corry, Wynn and Russell. Although it took some years for other labs to become ...

An Immunological Model for Heterotopic Heart and Cardiac Muscle Cell Transplantation in Rats

Heterotopic heart transplantation in rats has been a commonly used model for diverse immunological studies for more than 50 years. Several modifications have been reported since the first description in 1964. After 30 years of performing heterotopic heart transplantation in rats, we have developed a simplified surgical approach, which can be easily taught and performed without further surgical training or background.
After dissection of the ascending aorta and the pulmonary artery and ligation of superior and inferior caval and pulmonary veins, the donor heart is harvested and subsequently perfused with ice-cold saline solution supplemented with heparin. After clamping and incising the recipient abdominal vessels, the donor ascending aorta and pulmonary artery are anastomosed to the recipient abdominal aorta and inferior vena cava, respectively, using continuous running sutures.
Depending on different donor-recipient combinations, this model allows analyses of either acute or chronic rejection of allografts. The immunological significance of this model is further enhanced by a novel approach of in-ear injection of vital cardiac muscle cells and subsequent analysis of draining cervical lymphatic tissue.

We describe a model of heterotopic abdominal heart transplantation in rats, implying modifications of current strategies, which lead to a simplified surgical approach. Additionally, we describe a novel rejection model by in-ear injection of vital cardiac muscle cells, allowing further transplant immunological analyses in rats.

쥐에 있는 이종 심혼 및 심장 근육 세포 이식을 위한 면역학 모형

Heterotopic heart transplantation in rats has been a commonly used model for diverse immunological studies for more than 50 years. Several modifications ...

면역학 및 감염병학

Blood Circuit Reconstruction in an Abdominal Mouse Heart Transplantation Model

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the established technique for a modified blood circuit reconstruction in a heterotopic abdominal heart transplantation model is presented. This method uses the intrathoracic inferior vena cava (IIVC) instead of the pulmonary artery of the donor heart for the anastomosis to the inferior vena cava of the recipient. It is facilitating and improving success rates for abdominal heart transplantation in mice.

A novel technique for blood circuit reconstruction in a heterotopic abdominal mouse heart transplantation model is demonstrated.

복부 마우스 심혼 이식 모형에 있는 혈액 회로 재건

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the ...

A Simplified Model for Heterotopic Heart Valve Transplantation in Rodents

There is an urgent clinical need for heart valve replacements that can grow in children. Heart valve transplantation is proposed as a new type of transplant with the potential to deliver durable heart valves capable of somatic growth with no requirement for anticoagulation. However, the immunobiology of heart valve transplants remains unexplored, highlighting the need for animal models to study this new type of transplant. Previous rat models for heterotopic aortic valve transplantation into the abdominal aorta have been described, though they are technically challenging and costly. For addressing this challenge, a renal subcapsular transplant model was developed in rodents as a practical and more straightforward method for studying heart valve transplant immunobiology. In this model, a single aortic valve leaflet is harvested and inserted into the renal subcapsular space. The kidney is easily accessible, and the transplanted tissue is securely contained in a subcapsular space that is well vascularized and can accommodate a variety of tissue sizes. Furthermore, because a single rat can provide three donor aortic leaflets and a single kidney can provide multiple sites for transplanted tissue, fewer rats are required for a given study. Here, the transplantation technique is described, providing a significant step forward in studying the transplant immunology of heart valve transplantation.

This protocol describes a simple and efficient method for the transplantation of aortic valve leaflets under the renal capsule to allow for the study of alloreactivity of heart valves.

설치류의 이종 주제 심장 판막 이식을위한 단순화 된 모델

There is an urgent clinical need for heart valve replacements that can grow in children. Heart valve transplantation is proposed as a new type of ...

A Modified Cuff Technique for Mouse Cervical Heterotopic Heart Transplantation Model

Cardiac allograft rejection limits the long-term survival of patients after heart transplantation. A mouse heart transplantation model is ideal for investigating the mechanism of cardiac allograft rejection in preclinical studies because of their high homology with human genes. This understanding would help develop unique approaches to improving patients&#39; long-term survival treated with cardiac allografts. In a mouse model, abdominal donor heart implantation is commonly performed with an end-to-end&#160;anastomosis to the recipient&#39;s aorta and inferior vena cava using stitches. In this model, the donor&#39;s heart is implanted by end-to-end anastomosis to the recipient&#39;s carotid artery and jugular vein by the modified-Cuff technique. The transplantation surgery is performed without stitching and thus may increase the survival of the recipient since there is no interference with the blood supply and venous reflux of the lower body. This mouse model would help investigate the mechanisms underlying the immunological and pathological (acute/chronic) rejection of cardiac allografts.

In the present protocol, a mouse heart transplantation model is used for investigating the mechanism of cardiac allograft rejection. In this heterotopic heart transplantation model, operation efficiency is improved, and the survival of cardiac grafts is ensured by a cervical end-to-end anastomosis of heart implantation using a modified Cuff technique.

마우스 자궁경부 이종 심장 이식 모델을 위한 수정된 커프 기술

Cardiac allograft rejection limits the long-term survival of patients after heart transplantation. A mouse heart transplantation model is ideal for ...

A Heterotopic Rat Heart Transplantation Model using Circulatory Death Donor Hearts

The objective of this protocol is to set up a rat heterotopic heart transplantation model with donation after circulatory death (DCD) donor hearts. There are two setups for this protocol: heart donor setup and recipient setup. In the heart donor setup, Sprague Dawley rats are anesthetized, endotracheally intubated, and ventilated. The right carotid artery is cannulated to deliver heparin and the paralytic agent vecuronium-bromide. The DCD process is initiated by terminating the ventilation. After 20 min, the heart is exposed and the aorta distal to the brachiocephalic branch is clamped. At 25 min from terminating the ventilator, ice-cold University of Wisconsin (UW) solution is perfused through the carotid catheter to flush the heart. The heart is procured by dividing the aorta, pulmonary artery, venae cavae, and pulmonary veins and stored in UW solution for implantation. In the recipient setup, the Lewis rat is anesthetized with isoflurane. Slow-release buprenorphine is administered subcutaneously to facilitate a smooth postoperative recovery. Through a midline abdominal incision, the infra-renal aorta and the inferior vena cava are isolated and clamped with an atraumatic vascular clamp. The donor heart aorta and pulmonary artery are sutured to the recipient abdominal aorta and vena cava, respectively, with a running 8-0 Prolene. The vascular clamp is removed to reperfuse the heart. The abdominal wall is closed and the rat is recovered. After a set interval (24 h to 2 weeks), the recipient rat is anesthetized, the transplanted heart is exposed, and a balloon-tip-catheter is inserted into the left ventricle via the apex to record developed pressure and dP/dt using a data acquisition system. The heart tissue is collected for histology, immunology, or molecular analysis. A successful DCD donor rat heart transplantation model will allow further studies on the cardioprotective approaches to improve heart transplantation outcomes from DCD donors.

Here, we describe the preparation and technical details of murine heterotopic heart transplantation utilizing a circulatory death donor heart.

The objective of this protocol is to set up a rat heterotopic heart transplantation model with donation after circulatory death (DCD) donor hearts. There ...

Rat Model of Normothermic Ex-Situ Perfused Heterotopic Heart Transplantation

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

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

Normothermic Ex-Situ 관류 이종 심장 이식의 쥐 모델

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

Modified Heterotopic Abdominal Heart Transplantation and a Novel Aortic Regurgitation Model in Rats

Over the past 50 years, many researchers have reported heterotopic abdominal heart transplantation in mice and rats, with some variations in the surgical technique. Modifying the transplantation procedure to strengthen the myocardial protection could prolong the ischemia time while preserving the donor&#39;s cardiac function. This technique&#39;s key points are as follows: transecting the donor&#39;s abdominal aorta before harvesting to unload the donor&#39;s heart; perfusing the donor&#39;s coronary arteries with a cold cardioplegic solution; and topical cooling of the donor&#39;s heart during the anastomosis procedure. Consequently, since this procedure prolongs the acceptable ischemia time, beginners can easily perform it and achieve a high success rate.
Moreover, a new aortic regurgitation (AR) model was established in this work using a technique different from the existing one,&#160;which is created by inserting a catheter from the right carotid artery and puncturing the native aortic valve under continuous echocardiographic guidance. A heterotopic abdominal heart transplantation was performed using the novel AR model. In the protocol, after the donor&#39;s heart is harvested, a stiff guidewire is inserted into the donor&#39;s brachiocephalic artery and advanced toward the aortic root. The aortic valve is punctured by pushing the guidewire further even after the resistance is felt, thus inducing AR. It is easier to damage the aortic valve using this method than with the procedure described in the conventional AR model. Additionally, this novel AR model does not contribute to the recipient&#39;s circulation; therefore, this method is expected to produce a more severe AR model than the conventional procedure.

This study demonstrates a reproducible heterotopic abdominal heart transplantation technique in rats that beginners can learn and perform. Additionally, a novel aortic regurgitation model in rats is generated by performing heterotopic abdominal heart transplantation and damaging the donor's aortic valve using a guidewire after harvesting.

쥐의 수정된 이종 복부 심장 이식 및 새로운 대동맥 역류 모델

Over the past 50 years, many researchers have reported heterotopic abdominal heart transplantation in mice and rats, with some variations in the surgical ...