본 연구는 &quot;도이치 Forschungsgemeinschaft&quot;(DFG)에 의해 투자되었다. K. Schleimer는 노르 트라 인 베스트 팔렌의 리제 마이트 과학 기술부의 친목과 연구에 의해 지원되었다. 우리는 그녀의 뛰어난 편집 지원을 부인 메리 조안 Blümich에게 감사의 말씀을 전합니다. 박사 마리아 Kokozidou이는 Fresenius 2011_A61에서 자금을 조달했다.

1 실험 동물 <ol><li> 동물 보호 FELASA의 지침에 따라 수행 될 모든 실험 지역 윤리위원회의 승인 (K 40, 99분의 17)을 얻습니다. </li><li> (예를 들면, 찰스 강 WIGA GmbH의 Sulzfeld, 독일)의 상업적 개종에서 구매하는 남성 루이스 쥐를 사용할 수 있습니다. 여기에, 224 동물 (112 기증자, 112받는 사람) 112 이식을 수행하는 데 사용되었다. 받는 사람은 수술 후 육주에서 희생되었다. 동작시에, 동물은 250~350그램 무게한다. </li><li> 음식과 물을 무료로 이용할와 기후의 방에있는 동물이 오지 않게하십시오. </li></ol> 2 마취 및 수술 준비 <ol><li> 1 L / 분의 산소 유속, 및 마취를 유도하는 5 % 마취 가스와 함께 기화기 이소 플루오 란 마취 기계에 접속 플렉시 글라스 상자에 래트를 배치했다. 1.5 % 이소 플루오 란 마취를 유지한다. </li><li> 에서 쥐를 놓습니다온난화 접시에 누운 자세. </li><li> 제모 크림과 함께 복부의 털을 제거합니다. </li><li> 다음과 같은 무균 수술 부위 준비 : 벤치 탑을 청소 소독제를 사용합니다. 제 절개 사이트 이용 클로르헥시딘 소독, 알코올 및 제 최종적 스크럽 용액 (예컨대, 클로르헥시딘, 70 % 이소 프로필 알콜)을 참조. 일회용 수술 가운 및 수술 장갑을 착용 할 것. </li><li> 멸균 미세 악기를 사용합니다. </li><li> 12 ~ 20 배의 배율 동작 현미경을 사용합니다. </li></ol> 3 수술 <ol><li> 기부자 작업 준비 <ol><li> 중간 개복술을 수행하고 왼쪽 측면 로브와 히긴스에 의해 설명 된대로 이식의 중간 엽 절제. (14) </li><li> 그런 다음 hepatoduodenal 인대를 준비합니다. 우선 작은 담관의 혼잡과 팽창을 유도하기 위해 담관을 결찰. 그런 다음 결찰 및 gastroduode을 가로로 쪼개다NAL 정맥과 동맥. 그 후, 복강 트렁크 및 간동맥을 격리하며 왼쪽 위 동맥 비장 동맥을 가로로 쪼개다. 혼잡 한 담관을 절개하고, 그것으로 20-G 스텐트를 소개합니다. 스텐트의 위치는 전체 문부 담관 시스템의 배수되도록한다. 스텐트 원위부 담관을 가로로 쪼개다. </li><li> 바로 오른쪽 신장 정맥 위의 caval 정맥의 infrahepatic 세그먼트를 격리합니다. 관류 </li><li> 체강 트렁크 상기 대동맥 결찰 후 간문맥 클램프하고 절개. 포털 정맥 관류 카테터 (14 G)를 소개하고, 히스티딘 트립토판 - 케 토글 루타 레이트 (HTK) -solution의 30 ㎖ (12cm의 H2O, 4 ° C)로 관류를 수행합니다. </li><li> 다이아 프램 및 관류가 유출 할 수 있도록 suprahepatic caval 정맥을 잘라. </li><li> 대동맥 패치와 함께 복강 트렁크를 절제하고 히스티딘 트립토판 - 케 토글 루타 레이트 용액으로 씻어냅니다. 적출 </li><li> 바로 오른쪽 신장 정맥 위의 infrahepatic caval 정맥을 가로로 쪼개다. </li><li> 그리고, 문맥에서 0.3 mm의 직경을 갖는 스텐트를 배치하고 봉합사로 고정한다. 다음 동작 동안, 간문맥는 오른쪽 신장 동맥을 통해 동맥 화된다. 스텐트에 말단 포털 정맥을 가로로 쪼개다. </li><li> suprahepatic caval 정맥 결찰 한 후, 다이아 프램의 커프스와 함께 그것을 가로로 쪼개다. </li><li> 이식 절편과 얼음에 비닐 봉지에 4 ° 섭씨 히스티딘 트립토판 - 케 토글 루타 레이트 (HTK) -solution에 보관합니다. infrahepatic caval 정맥 장소 코너 봉합 (8-0 Ethilon). </li></ol></li><li> 받는 사람 운영 <ol><li> 중간 개복술을 수행하고 infrahepatic caval 정맥을 준비합니다. 그리고, 오른쪽 신장 동맥 및 요관을 분리 요관을 결찰하고 쪼개다. 그 기원에서 오른쪽 신장 동맥 클램프, 다음 오른쪽 신장 정맥 결찰. 그리고 모두의 절개 후 다시NAL 혈관, 신장 절제술을 수행합니다. 이어서, 오른쪽 신장 동맥을 단축 헤파린 식염수 (40 UI 헤파린 / ml의 생리 식염수 용액)로 헹군다. </li><li> infrahepatic caval 정맥 타원형 루멘을 만들 infrahepatic caval 정맥의 근위 및 원위 체결 후 합자에 근위 오른쪽 신장 정맥을 가로로 쪼개다. 헤파린 생리 식염수로 caval 정맥을 씻어. 받는 사람의 복부의 오른쪽 신장 지역에서 이식편을 이식. </li><li> 8-0 Ethilon 실행 봉합 recipient`s caval 정맥에 기증자의 infrahepatic caval 정맥 엔드 - 투 - 사이드 Anastomose. </li><li> 헤파린 / 식염수로 세정 한 후, 오른쪽 신장 동맥 (도 3)로 문맥에 스텐트를 소개한다. 이 바늘 (8-0 Ethilon)와 포털 정맥 및 신장 동맥을 연결합니다. 합자 (꼰 실크 블랙 7-0)로 신장 동맥에 연결 스텐트를 고정합니다. </li><li> 재관류를 허용하려면 먼저 모두 다음, 동맥 클램프를 제거정맥 클램프 (그림 4). </li><li> infrarenal 대동맥을 준비하고 근위 및 원위 microclamp를 배치하고 오른쪽 두 클램프 사이에 대동맥을 절개. 10-0 Ethilon 실행 봉합, 대동맥 패치를 사용하여 대동맥과 기증자의 복강 트렁크 사이의 엔드 - 투 - 측 문합을 수행합니다. </li><li> 십이지장으로 담관을 삽입 : 십이지장 벽에 pursestring 봉합을 적용, 오른쪽 pursestring의 중간에 십이지장 벽에 작은 절개를합니다. 십이지장에 스텐트 담관을 소개하고, pursestring 봉합사를 조입니다. 마지막으로, 단단히 두 개의 바늘 (Ethilon 8-0) (그림 5)와 십이지장과 담도를 개최합니다. </li><li> 그런 다음 기본 간 절제술, 첫째 Emond 등으로 설명을 수행합니다. 급성 간부전을 모방합니다. 7 </li><li> 다시의 측면 복벽에 함께 다이어프램의 팔목, 이식편의 suprahepatic caval 정맥 수정4-0 Vicryl 봉합사와 cipient. 생리 식염수 (그림 6)과 복부를 씻어. </li><li> 근육 층은 4-0 Vicryl 실행 봉합하고 피부에 3-0 Vicryl 실행 봉합을 사용합니다. </li></ol></li></ol> 4 복구 및 수술 후 관리 <ol><li> 동물 적외선 온난화 램프에서 마취에서 회복하자. </li><li> 이일에 대한 0.03-0.05 ㎎ / ㎏ BW SC마다 8 ~ 12 시간의 용량으로 부 프레 놀핀을 주입한다. </li><li> 일반 상태와 질병이나 감염의 징후에 관한 동물을 검사합니다. </li></ol> 초기 기본 간 무게의 5 계산 다음과 같이 초기 원시 간 중량을 계산한다 : 완전한 간 중량로부터 절제 간 로브의 중량을 뺀다 (체중 (+/- 0.49 %의 표준 편차가이 값의 2.66 %가 N과 이전의 실험에서 얻은 = 같은 변형과의 80 동물GE의)). 15

<ol>
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모든 저자는 경쟁 이익을 선언하지 않습니다.

제 이소성 보조 쥐 간 이식가 Lee 등으로 기술되었다., 1966 년 16 헤스 하였다. 그래프트 17 간문맥은 수신자의 간문맥으로 문합 하였다. 이 절차 동안, 네이티브간에 포털 정맥 혈액 공급이 저하된다. 1991 년 홍콩은 혈액의 흐름 조절 대규모 포털 않고.받는 사람의 장골 동맥을 통해 PVA와 13 HALT를 수행 정현파 정체가 발생했습니다. <p class="jove_co...

급성 간부전은 60~80%의 사망률. 8 급성 간부전에서 선도 임상 의학에서 심각한 문제가있다, 보조 간 이식은 급성기를 해소 할 수있는 유망한 대체 요법이다. 기본 간은 면역 억제 약물을 중단 할 수 있습니다 재생에 다음 재생성를 유지한다. 결과적으로 이식이 위축 될 것이다. 그러나 보조 간 이식을위한 수술 기법이 여전히 논의된다. 1-3 이러한 맥락에서, 테르 프 스트라가 그래프트 공간의 부족을보고, 포털 혈액 현상 및 정맥 울혈 및 배수의 문제를 훔칠. 9-10 도너의 간문맥 받는 사람의 문맥에 엔드 - 투 - 측 문합 하였다. 이것은 이식하고 원시 간 사이 포털 정맥 혈전증 포털 흐름 경쟁의 결과. 이러한 기술적 문제는 대부분 APOLT-방법의 사용 (orthotop으로 회피 할 수있다IC 보조 세그먼트의 주입). 4 그러나이 방법은 원시 간 이식. 4 1998 년 모두의 광범위한 절제를 필요로, 에어 하드와 그의 팀은 포털 정맥 arterialization (PVA) (그림 1을 이용하여 이소성 보조 간 이식 (HALT)을 개발 ). 폐문하고 원시 간 문맥은이 방법의 사용으로 존경합니다. 즉, 원시 간 회복을 방해 할 수있는 등 포털 정맥 혈전증 및 포털 혈액과 같은 원치 않는 부작용에서받는 사람 훔치는 현상을 여분. 또한, 네이티브 간 이식의 더 광범위한 간 절제술은 (이식) 및 재생 (기본 간)을위한 연계 결과적으로 더 적은 출혈이 더 남아있는 총 간 실질 질량이 방법에 대한 필요하지 않습니다. 이 기술은 초기 임상 결과를 약속 하였다. 5-6 일이 실험의 목적은 간문맥 arterialization에서와 HALT 기술의 개발이었다연구 목적으로 쥐 (도 2). 예비 실험에서 우리는 간 캡슐에서 재관류 및 출혈 후 이식 이식의 혼잡 초래하는 정맥 고려 동맥 화 포털의 혈류 조절을하지 않았다. 11 동맥 화 문맥에서 다음 혈액의 흐름 때문에 0.3 mm의 내부 직경 8 mm의 길이를 가진 스텐트 의해 조절 하였다. 파일럿 실험은 0.3 mm의 스텐트 직경이 상부 생리적 범위 동맥 화 간문맥의 평균 혈류를 얻을 수 있음을 보여 주었다 (1.7 +/- 0.4 ㎖ / 분 / g 간 중량). 혈류 조절없이 12 이상의 결과 혈액의 흐름 조절 대규모 포털 않고받는 사람의 장골 동맥을 통해 PVA와 함께 자신의 실험에서. 13 HALT를 수행 정현파 혼잡이 발생한 홍콩의 결과와 상관 관계. 비디오 우리의 목적은 D이다단계별로 emonstrate 단계 쥐에서이 ne.w 실험적인 수술 기법. 장기적으로 제안 된 기술은 간 재생 및 상호 간 경쟁과 간문맥 arterialization, 간문맥과 환류 및 포털 혈류 조절에 관한 연구에 사용될 수있다.

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

급성 간부전 보조 간 이식에 흥미있는 또 다른 방법이다. 목표는 실패 원시 간 재생 될 때까지 임시 지원을 제공하는 것입니다. 1-3 APOLT - 방법, 보조 segments-의 소성을 주입은 기술적 인 문제의 대부분을 피한다. 그러나이 방법은 원시 간 이식. 4 1998 년 모두의 광범위한 절제를 필요로한다, 에어 하드는 포털 정맥 arterialization (PVA) (그림 1)를 이용 이소성 보조 간 이식 (HALT)를 개발했다. 이 기술은 초기 임상 결과를 약속했다. 5-6 우리는 이식의 형태와 기능 (그림 2)에 유량 조절 PVA의 영향을 조사하기 위해 쥐의 흐름을 조절 PVA와 HALT-기술을 개발했다. 원래 크기의 30 %로 감소 간 이식은 heterotopically 오른쪽 신장 적출 후 수신자의 오른쪽 신장 지역에 주입 하였다.60; 이식의 적외선 간 caval 정맥받는 사람의 infrahepatic caval 정맥과 문합 하였다. 기증자의 문맥의 arterialization은 스텐트 기술로받는 사람의 권리 신장 동맥을 통해 실시 하였다. 동맥 화 간문맥의 혈류 조절은 0.3 mm의 내부 직경을 갖는 스텐트를 이용하여 달성되었다. 이식의 복강 트렁크는 엔드 - 투 - 사이드받는 사람의 대동맥과 담관과 문합은 십이지장에 이식 하였다. 원시 간 절제술 급성 간부전을 유도 하였다. 7 이러한 방식으로 112 이식을 시행 하였다. 수술 후 생존율은 90 %이었고, 6 주간의 생존율은 80 %이었다. 6 주 수술 후, 기본 간은 9.8 ± 1g에 2.3 ± 0.8 g 중량의 증가를 보여주는, 재생. 이때, 그래프트의 중량을 2.3 ± 0.8 g에 3.3 ± 0.8 g 감소 하였다. </p&gt; 우리는 그래프트 형태와 기능의 관점에서 유망 장기적인 결과를 얻을 수 있었다. 기본 간은 다시 생성 될 때까지 흐름을 조절 PVA와 HALT 안정적으로 급성 간부전 교량.

이식의 평균 지속 시간은 약 150 분이었다. 냉 허혈 시간은 64 분이었다 +/- 7 따뜻한 허혈 시간은 25 분 미만이었다. 수술 후 생존율은 90 %이었고, 6 주간의 생존율은 80 %이었다. 20 % 손실은 첫번째 수술 후 주 동안 때문에 스텐트 전위 및 담즙 누출 (네 동물)의 복막염에 의한이었고, 다음과 같은 이유로 오주 동안 폐렴, 복강 내 농양 및 장폐색증 (일곱 동물)이다. 살아남은 동물은...

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

보조 간 이식은 실패 간 재생 될 때까지, 급성 간부전에 임시 지원을 제공합니다. 포털 정맥 arterialization (PVA)와 이소성 보조 간 이식 (HALT)는 충분한 간 기능을 렌더링합니다. 우리는 이식의 형태와 기능에 대한 포털 정맥 arterialization의 영향을 조사하기 위해, 쥐에서 유사한 기술을 개발했다.

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 ...

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

Research

JoVE Journal

Medicine

12.6K 조회수.  University Hospital RWTH Aachen.  보조 간 이식은 실패 간 재생 될 때까지, 급성 간부전에 임시 지원을 제공합니다. 포털 정맥 arterialization (PVA)와 이소성 보조 간 이식 (HALT)는 충분한 간 기능을 렌더링합니다. 우리는 이식의 형태와 기능에 대한 포털 정맥 arterialization의 영향을 조사하기 위해, 쥐에서 유사한 기술을 개발했다. 

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

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

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

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 ...

연구

의학

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.

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 ...

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.

마우스의 이소성 자궁 심장 이식

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

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

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

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

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

연결 간 파티션과 무대 Hepatectomy (ALPPS) 절차에 대 한 포털 정 맥 결 찰의 쥐 모델

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.

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. ...

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.

포털-고혈압 증후군 Cirrhotic 쥐에서의 침입 Hemodynamic 특성화

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% 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 ...

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

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

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

마우스에서 간내 섬 이식 중 주입 후 포털 정맥 출혈 최소화

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

Rat Model of Normothermic Ex-Situ Perfused Heterotopic Heart Transplantation

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

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

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 ...

내피에서 중간엽으로의 전환을 연구하기 위한 생체 내 심장 이식

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

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

저자 스포트라이트: 내피에서 중간엽으로의 전환 연구를 위한 신생아 이종성 쥐 심장 이식 모델  

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

내피에서 중간엽으로의 전환 연구를 위한 신생아 이종성 쥐 심장 이식 모델

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