이 작품은 CKB에 NIH (RO1 HL098228)에서 교부금에 의해 부분적으로 지원되었다.

참고 : 모든 동물의 절차는 전국 어린이 병원 기관 동물 관리 및 사용위원회에 의해 승인되었습니다. 심장 밸브 기증자 마우스 1. 폐 심장 밸브 수확 <ol><li> 수술 전에 수술 도구를 압력솥 : 미세 가위, 배 마이크로 집게, 배 미세 혈관 클램프, 1X 클램프 적용 집게, 1X 마이크로 니들 홀더, 1X 봄 가위, 1X 트랙터를 1x는. </li><li> 6-8 주 된 여성 C57BL / 6 마​​우스는 폐 심장 판막 기증자로 사용됩니다. 케이지에서 마우스를 제거하고 무게를 달고, 다음 케타민 / 자일 라진 칵테일 (케타민, 200 ㎎ / ㎏과 자일 라진, 20 ㎎ / kg, IP) 과다 복용으로 안락사. </li><li> 가슴 부분을 클립 및 패드에 지느러미 드러 누움 위치에 마우스를 놓습니다. 그런 다음 개흉술을합니다. 심장을 노출 우심방에 작은 상처를 확인하고 차가운 얼음 식염수 좌심실을 perfuse. </li><li> 퉁명스럽게 상승에서 폐동맥 (PA)를 해부하다오르타. 폐동맥의 2mm의 커프스와 함께 폐 밸브 (PV)를 잘라. 마음의 나머지를 폐기하십시오. </li><li> 차가운 헤파린과 생리 식염수 (100 단위 / ml)에 PV를 저장합니다. 주 : PV는 기증자의 심장에 이식하기 전에 2 시간 동안 용액에 보관하실 수 있습니다. </li></ol> 2. 기증자의 심장 준비 <ol><li> 6-8 주 된 여성 C57BL / 6 마​​우스가 심장 기증자로 사용됩니다. 케이지에서 마우스를 제거하고 무게를 달고, 다음 케타민 / 자일 라진 칵테일 (케타민, 200 ㎎ / ㎏과 자일 라진, 20 ㎎ / kg, IP) 과다 복용으로 안락사. 이 터미널의 절차입니다. </li><li> 가슴 부분을 클립 및 패드에 지느러미 드러 누움 위치에 마우스를 놓습니다. 그런 다음 개흉술을합니다. 퉁명스럽게 심장, 하대 정맥 (IVC), 우수한 대정 맥 (SVC), 상행 대동맥, PA, 폐 정맥을 분리합니다. 얼음 차가운 멸균 식염수로 IVC를 Perfuse. </li><li> IVC, SVC하고 우수한 잘라 6-0 실크 봉합사와 폐 혈관을 결찰합자. </li><li> 2mm 팔목과 대동맥 및 PA를 잘라. </li><li> PV를 잘라서 폐기하십시오. </li></ol> 기증자의 심장 위에 3. 심장 밸브 이식 <ol><li> 즉시 2.5 단계 후, 기증자의 마음에 단계 1.5에서 심장 밸브를 배치하고 심장 밸브의 방향. </li><li> 테이퍼 바늘에 10-0 모노 필라멘트 봉합사를 사용하여 밸브의 오른쪽에 스티치와 PV를 확보하고 태양 광 발전의 다른 측면에서 5-6 stiches와 함께 지속적으로 봉합하기 시작합니다. </li><li> 전방 측을 마친 후 수평 심장 회전 공여체 심장에 PV의이면을 봉합하기 시작한다. </li><li> 차가운 멸균 헤파린 / 식염수에 마음을 저장합니다. 참고 : 기증자의 마음이받는 마우스에 주입하기 전에 2 시간 동안 용액에 보관하실 수 있습니다. </li></ol> 받는 사람 마우스에 4. 이소성 심장 이식 <ol><li> 6-8 주 된 여성 C57BL / 6 마​​우스 RECI로 사용되었다pient. 케이지에서 마우스를 제거하고 케타민 / 자일 라진 칵테일 (케타민, 100 ㎎ / ㎏과 자일 라진 10 ㎎ / ㎏)으로 마취, 그것은 무게. 케토 프로 펜 (5 ㎎ / kg)을 preanesthesia 진통제로서 사용된다. </li><li> 곤란 꼬리에 의한 진정 작용의 수준을 확인한 후, 복부와 가슴 머리를 클립. 멸균 안과 연고와 함께 눈에 기름칠을하고, 패드에 지느러미 드러 누움 위치에 마우스를 놓습니다. 베타 딘, 알코올 패드로 복부를 소독. 그런 다음 멸균 드레이프와 마우스를 커버 만 절개 영역을 노출합니다. </li><li> 치골 지역에 xyphoid 아래에서 중간 선 개복술의 절개를하고, 자체 유지 견인기를 삽입합니다. 식염수를 적신 거즈에 내장을 감싸십시오. 퉁명스럽게 신동맥 대동맥과 대정맥을 정의합니다. </li><li> 혈액 순환을 억제하는 대동맥과 IVC 주변의 근위 및 원위 두 6-0 실크 봉합사를 놓습니다. </li><li> 복부 대동맥의 오른쪽에있는 기증자의 심장을 놓고 멸균 GAU로 커버&#39;제. 식염수로 보습. </li><li> 30 G 바늘을 사용하여 복부 대동맥에 aortotomy을 확인하고 기증자의 대동맥의 크기에 가위로 구멍을 확장합니다. </li><li> 테이퍼 바늘에 멸균 10-0 모노 필라멘트 봉합사를 사용하여 측면 엔드 - 투 문합을 수행합니다. 복부 대동맥의 입구의 기단에 하나의 스티치 기증자의 대동맥을 확보하고 복부 대동맥의 말단부에서 4-5 stiches와 함께 지속적으로 봉합하기 시작합니다. </li><li> 왼쪽에 마음을 뒤집기 식염수 주입 거즈로 커버하고, 복부 대동맥의 말단부에서 4-5 stiches와 지속적으로 봉합하기 시작합니다. </li><li> 30 G 바늘을 사용하여 IVC의 venotomy을 확인하고 기증자 폐동맥의 크기로 구멍을 확장합니다. </li><li> 테이퍼 바늘에 멸균 10-0 모노 필라멘트 봉합사를 사용하여 측 문합에 종지부를 수행합니다. IVC의 개구부의 기단에 하나의 스티치 기증자 PA를 확보에서 4-5 stiches를 연속 봉합 시작하대 정맥의 말단. 대동맥 길에 있기 때문에 이번에는 반드시 기증자의 PA의 왼쪽 벽의 봉합은 IVC의 내부에 있는지 확인합니다. </li><li> 헤파린과 생리 식염수 (100 단위 / ml)로 I​​VC 루멘을 세척합니다. 선단에 계속을 봉합하여 기증자 PA 사람과받는 사람 IVC의 오른쪽 벽을 닫습니다. </li><li> 원심 합자를 제거하고 국소 흡수 멸균 지혈 에이전트를 적용하여 출혈을 제어 할 수 있습니다. 출혈이 완전히 정지 할 때, 근위 봉합사를 제거하고 출혈 같은 방법으로 제어 할 수 있습니다. </li><li> 창자를 반환하고 6-0 블랙 폴리 아미드 모노 필라멘트 봉합사를 사용하여 두 층으로 복부 근육과 피부를 닫습니다. </li><li> 0.5 ㎖의 생리 식염수를 피하 주사와 마우스가 완전히 이동할 때까지 온난화 패드 복구 새장에 마우스를 놓습니다. 복구시, 종이 침구 새 케이지에 마우스를 반환합니다. 48 시간 동안 진통제 (이부프로펜, 30 ㎎ / ㎏, 식수를) 제공합니다. 할완전히 회복 될 때까지 다른 동물의 회사에 수술을했다 동물을 반환하지. </li></ol>

<ol>
	<li>Polito, 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=Increased+morbidity+and+mortality+in+very+preterm+VLBW+infants+with+congenital+heart+disease.">Increased morbidity and mortality in very preterm VLBW infants with congenital heart disease.</a> Intens Care Med. 39, 1104-1112 (2013).</li><li>Wren, C., Reinhardt, Z., Khawaja, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Twenty+year+trends+in+diagnosis+of+life+threatening+neonatal+cardiovascular+malformations.">Twenty year trends in diagnosis of life threatening neonatal cardiovascular malformations.</a> Arch Dis Child Fetal. 93, F33-F35 (2008).</li><li>Vacanti, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beyond+transplantation+Third+annual+Samuel+Jason+Mixter+lecture.">Beyond transplantation Third annual Samuel Jason Mixter lecture.</a> Archives of surgery. 123, 545-549 (1960).</li><li>Tudorache, I., 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=Orthotopic+replacement+of+aortic+heart+valves+with+tissue-engineered+grafts.">Orthotopic replacement of aortic heart valves with tissue-engineered grafts.</a> Tissue engineering Part A. 19, 1686-1694 (2013).</li><li>van Geldorp, M. W., 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=Patient+outcome+after+aortic+valve+replacement+with+a+mechanical+or+biological+prosthesis+weighing+lifetime+anticoagulant+related+event+risk+against+reoperation+risk.">Patient outcome after aortic valve replacement with a mechanical or biological prosthesis weighing lifetime anticoagulant related event risk against reoperation risk.</a> The Journal of thoracic and cardiovascular surgery. 137, 881-886 (2009).</li><li>El Oakley, R., Kleine, P., Bach, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Choice+of+prosthetic+heart+valve+in+today's+practice.">Choice of prosthetic heart valve in today's practice.</a> Circulation. 117, 253-256 (2008).</li><li>Quinn, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Models+for+Bench+to+Bedside+Translation+of+Regenerative+Cardiac+Constructs.">Animal Models for Bench to Bedside Translation of Regenerative Cardiac Constructs.</a> Progress in Pediatric cardiology. 35, (2013).</li><li>Shinoka, T., 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=Tissue+engineering+heart+valves+valve+leaflet+replacement+study+in+a+lamb+model.">Tissue engineering heart valves valve leaflet replacement study in a lamb model.</a> The Annals of thoracic surgery. 60, S513-S516 (1995).</li><li>Sodian, R., 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=Tissue+engineering+of+heart+valves+in+vitro+experiences.">Tissue engineering of heart valves in vitro experiences.</a> The Annals of thoracic surgery. 70, 140-144 (2000).</li><li>Shinoka, T., 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=Tissue+engineered+heart+valves.+Autologous+valve+leaflet+replacement+study+in+a+lamb+model.">Tissue engineered heart valves. Autologous valve leaflet replacement study in a lamb model.</a> Circulation. 94, II164-II168 (1996).</li><li>Sodian, R., 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=Evaluation+of+biodegradable+three+dimensional+matrices+for+tissue+engineering+of+heart+valves.">Evaluation of biodegradable three dimensional matrices for tissue engineering of heart valves.</a> ASAIO journal (American Society for Artificial Internal Organs. 46, 107-110 (1992).</li><li>Sodian, R., 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=Early+in+vivo+experience+with+tissue-engineered+trileaflet+heart+valves).">Early in vivo experience with tissue-engineered trileaflet heart valves).</a> Circulation. 102, III22-III29 (2000).</li></ol>

우리는 공개 아무것도 없어.

이 절차의 사망률은 주로받는 사람 복부 대동맥에 기증자의 대동맥의 PV 이식 사이트와 문합에서 출혈에 의해 발생 된 20 %에 가깝습니다. 대부분의 경우, 사망률이 유의하게 48 시간의 수술 후 감소한다. 생존 마우스는 이식 된 태양 광 발전을 통해 강한 심장 박동, 혈액 흐름을 보여 주었다. 전체 과정은 경험이 풍부한 마이크로 외과 의사 네 시간이 소요됩니다. 이 기술을 마스터하기 위해 약 250 ?...

선천성 심장 결함은 서쪽 세계 1,2에서 유아 사망의 주요 원인 중 하나입니다. 그 중, 폐동맥 판막 협착과 뾰족한 끝이 둘있는 대동맥 판막 결함이 자주 발생하는 형태 3입니다. 심장 판막 교체 수술은 재건 수술의 일상적인 선택이 될 것입니다; 그러나, 협착 및 석회화 심장 밸브 및 항응고제에 평생 의존 등의 합병증은 만성 건강 쇠약과 죽음 4-7의 중요한 원천입니다. 또한, 성장 잠재력의 부족은 더욱 그 젊은 환자 4,8,9의 사망률을 증가시키는 개정 수술을 필요로합니다. 생분해 성 합성 심장 밸브 (8) 상에 성장 잠재력, Shinoka 외. 시드자가 세포와 기능적 여분 심장 밸브를 개발하기위한 시도. 성장 potenti와 같은 구조 네이티브 심장 밸브로 변환 합성 밸브알. 예비 많은 동물 연구는 기능적인 심장 밸브 (10)를 생성하기 위해이 방법을 사용할 수있는 가능성을 보여 주었다. 그러나, 장기 이식 연구 인해 심장 판막의 협착 발생 밸브 neotissue 점진적으로 두껍게별로 내구성을 보여 주었다. Sodian에서 작업 등. Shinoka 방법을 사용하지만, 궁극적으로 더 많은 생리 학적 프로파일 9,11,12을 구성 조직 설계 밸브의 생 역학적 특성을 준 생분해 성 엘라스토머와 PGA 매트릭스를 대체했다. 생체 내 연구에서는 이식의 성공에도 불구하고, 합류 내피 세포 라이닝이 지지체 (12)의 장기 성공을 제한 할 수있는 형성되지 않았다. 합리적 개선 번째 세대 합성 심장 밸브를 설계하기 위해, 심장 판막 이식 뮤린 모델은 세포 및 분자 메커니즘 underlyin을 조사하기 위해 만들어진G의 neotissue 형성, 밸브 두껍게, 그리고 협착 개발. 쥐 모델은 다른 종 7에서 쉽게 사용할 수없는 형질 전환 등의 분자 시약의 광대 한 배열을 제공하고 있습니다. 이 심장 판막 이식 모델에서 생체 동계 폐 심장 판막 치환술은 우선 수행 하였다; 다음 주입 심장 밸브와 심장은 미세 수술 기술을 이용하여 동계 호스트로 heterotopically 주입 하였다. 이 모델은, 체외 순환을위한 필요없이 심장 밸브 교체를 가능하게한다. 이 논문에서, 심장 판막 수확에 대한 자세한 설명은, 기증자의 심장 준비, 심장 판막 이식, 이소성 심장 이식이 설명되어 있습니다. 결과는받는 사람의 심장 박동의 독립적 인 한 기증자의 심장에서 연속적인 심장 박동을 보여 주었다. 주입 폐동맥 밸브를 통해 혈류는 펄 스파 명소와 고주파 초음파 시스템을 이용하여 측정 하였다ppler.

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			<td height="20" style="height:20px;width:176px;">DPBS</td>
			<td style="width:167px;">gibco</td>
			<td style="width:133px;">14190-144</td>
			<td style="width:168px;"></td>
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			<td height="24" style="height:24px;width:176px;">Microscope</td>
			<td style="width:167px;">Leica</td>
			<td style="width:133px;">M80</td>
			<td style="width:168px;"></td>
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		<tr height="25">
			<td height="25" style="height:25px;width:176px;">C57BL/6J (H-2b), Female</td>
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</table>

transplantation of pulmonary valve using a mouse model of heterotopic heart transplantation

조직 공학 심장 판막, 특히 decellularized 밸브, 혼합 결과와 교정 수술의 임상 사용에 추진력을 얻기 시작됩니다. 그러나, neotissue 개발, 밸브 두껍게, 그리고 협착 개발의 세포 및 분자 메커니즘은 광범위하게 연구되어 있지 않습니다. 위의 질문에 대답하기 위해, 우리는 쥐의 이소성 심장 판막 이식 모델을 개발했다. 심장 밸브는 밸브 기증자 마우스에서 수확 및 심장 기증자 마우스에 이식했다. 새로운 밸브 마음은받는 사람 마우스 heterotopically 이식했다. 이식 된 심장은받는 사람의 심장 박동의 독립적 인 자신의 심장 박동을 보여 주었다. 혈류는 펄 스파 도플러와 고주파 초음파 시스템을 이용하여 정량 하였다. 이식 된 폐 밸브를 통과하는 흐름은 최소한의 역류와 흐름을 앞으로 보여 피크 흐름은 100mm / 초에 가까웠다. 심장 판막 이식이 쥐 모델은 highl입니다다목적 Y, 그래서 수정 된 다른 혈역학 환경을 제공하기 위하여 및 / 또는 조직 공학 심장 판막에 neotissue 개발을 연구하기위한 다양한 트랜스 제닉 마우스와 함께 사용할 수있는 적응 될 수있다.

도 1은 이소성 심장 이식을 이용한 심장 판막 이식 모델의 개략도를 도시한다. 심장 판막은 기증자의 심장에서 수확 및 제 기증자 마우스의 심장에 이식했다. 그런 다음 새로운 심장 판막과 마음이받는 사람 마우스의 복부에 이식했다. 2 오른쪽 심장 이식 (B) 후, 복부 공간 (A)에 이식 마음의 그림을 보여주고, 이식 후 5 분 그림. 대동?...

Watch this Scientific Journal Video about Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation at JoVE.com

Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

조직 공학 심장 판막에 neotissue 형성 및 협착증 개발을 기초 세포 및 분자 메커니즘을 이해하기 위해, 이소성 심장 판막 이식 뮤린 모델이 개발되었다. 폐동맥 판막은 이소성 심장 이식 기술을 사용하여 수신자에게 이식 하였다.

이소성 심장 이식 마우스 모델을 사용하여 폐동맥 판막 이식

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed ...

transplantation-pulmonary-valve-using-mouse-model-heterotopic-heart

Research

JoVE Journal

Medicine

18.8K 조회수.  Nationwide Children's Hospital.  조직 공학 심장 판막에 neotissue 형성 및 협착증 개발을 기초 세포 및 분자 메커니즘을 이해하기 위해, 이소성 심장 판막 이식 뮤린 모델이 개발되었다. 폐동맥 판막은 이소성 심장 이식 기술을 사용하여 수신자에게 이식 하였다. 

비디오: Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

A Mouse Model of in Utero Transplantation

The transplantation of stem cells and viruses in utero has tremendous potential for treating congenital disorders in the human fetus. For example, in utero transplantation (IUT) of hematopoietic stem cells has been used to successfully treat patients with severe combined immunodeficiency.1,2 In several other conditions, however, IUT has been attempted without success.3 Given these mixed results, the availability of an efficient non-human model to study the biological sequelae of stem cell transplantation and gene therapy is critical to advance this field. We and others have used the mouse model of IUT to study factors affecting successful engraftment of in utero transplanted hematopoietic stem cells in both wild-type mice4-7 and those with genetic diseases.8,9 The fetal environment also offers considerable advantages for the success of in utero gene therapy. For example, the delivery of adenoviral10, adeno-associated viral10, retroviral11, and lentiviral vectors12,13 into the fetus has resulted in the transduction of multiple organs distant from the site of injection with long-term gene expression. in utero gene therapy may therefore be considered as a possible treatment strategy for single gene disorders such as muscular dystrophy or cystic fibrosis. Another potential advantage of IUT is the ability to induce immune tolerance to a specific antigen. As seen in mice with hemophilia, the introduction of Factor IX early in development results in tolerance to this protein.14 
In addition to its use in investigating potential human therapies, the mouse model of IUT can be a powerful tool to study basic questions in developmental and stem cell biology. For example, one can deliver various small molecules to induce or inhibit specific gene expression at defined gestational stages and manipulate developmental pathways. The impact of these alterations can be assessed at various timepoints after the initial transplantation. Furthermore, one can transplant pluripotent or lineage specific progenitor cells into the fetal environment to study stem cell differentiation in a non-irradiated and unperturbed host environment.
The mouse model of IUT has already provided numerous insights within the fields of immunology, and developmental and stem cell biology. In this video-based protocol, we describe a step-by-step approach to performing IUT in mouse fetuses and outline the critical steps and potential pitfalls of this technique.

A Mouse Model of in Utero Transplantation

The mouse model of in utero transplantation is a versatile tool that can be used to study the potential clinical applications of stem cell transplantation and gene therapy in the fetus. In this protocol, we present a general approach to performing this technique

의 마우스 모델 Utero에 이식

The transplantation of stem cells and viruses in utero has tremendous potential for treating congenital disorders in the human fetus.  For example, in ...

연구

의학

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are available to perform mechanistic in vivo studies. While heart transplantation models using a suture technique were first successfully developed in rats, the translation into an equally widespread used murine equivalent was never achieved due the technical complexity of the microsurgical procedure. In contrast, non-suture cuff techniques, also developed initially in rats, were successfully adapted for use in mice1-3. This technique for revascularization involves two major steps I) everting the recipient vessel over a polyethylene cuff; II) pulling the donor vessel over the formerly everted recipient vessel and holding it in place with a circumferential tie. This ensures a continuity of the endothelial layer, short operating time and very high patency rates4.
Using this technique for vascular anastomosis we performed more than 1,000 cervical heart transplants with an overall success rate of 95%. For arterial inflow the common carotid artery and the proximal aortic arch were anastomosed resulting in a retrograde perfusion of the transplanted heart. For venous drainage the pulmonary artery of the graft was anastomosed with the external jugular vein of the recipient5.
Herein, we provide additional details of this technique to supplement the video.

The murine cervical heart transplantation model is well suited for immunological as well as ischemia reperfusion injury studies. We modified the procedure using a non-suture cuff technique and performed more than 1,000 successful transplants with this approach.
Herein, we provide additional details of this technique to supplement the video.

수정 커프 기술을 사용하여 쥐의 자궁 심장 이식 모델

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are ...

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 ...

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 ...

A Novel Microsurgical Model for Heterotopic, En Bloc Chest Wall, Thymus, and Heart Transplantation in Mice

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression must be pursued. Employing vascularized bone marrow transplantation and co-transplantation of the thymus have shown promise in this regard in various animal models.1-11 Vascularized bone marrow transplantation allows for the uninterrupted transfer of donor bone marrow cells within the preserved donor microenvironment, and the incorporation of thymus tissue with vascularized bone marrow transplantation has shown to increase T-cell chimerism ultimately playing a supportive role in the induction of immune regulation. The combination of solid organ and vascularized composite allotransplantation can uniquely combine these strategies in the form of a novel transplant model. Murine models serve as an excellent paradigm to explore the mechanisms of acute and chronic rejection, chimerism, and tolerance induction, thus providing the foundation to propagate superior allograft survival strategies for larger animal models and future clinical application. Herein, we developed a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique. The experience in syngeneic and allogeneic transplant settings is described for future broader immunological investigations via an instructional manuscript and video supplement.

To study combined solid organ and vascularized composite allotransplantation, we describe a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique.

마우스의 소설 미세 수술 이소성 모델, 욕실 공산권 가슴 벽, 흉선, 및 심장 이식

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression ...

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure. Multiple small-animal models of heart transplantation have been used to study the acute and long-term effects of novel therapies. However, few are tested and demonstrated success in clinical trials. It is of critical importance to evaluate new therapies in a clinically relevant large-animal model for efficient and reliable translation of basic studies&#39;&#160;findings. Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and previously used to investigate novel cardioprotective strategies. This procedure focuses on acute ischemia-reperfusion injury and is a reliable method to investigate novel interventions which have been tested and validated in smaller experimental models, such as the murine model. We demonstrate its usefulness in assessing cardiac performance during the early post-transplantation period and other potential possibilities enabled by the model.

Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and utilized to investigate novel cardioprotective strategies.

정형외 심장 이식의 전 임상 돼지 모델

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart ...

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 Porcine Heterotopic Heart Transplantation Protocol for Delivery of Therapeutics to a Cardiac Allograft

Cardiac transplantation is the gold standard treatment for end-stage heart failure. However, it remains limited by the number of available donor hearts and complications such as primary graft dysfunction and graft rejection. The recent clinical use of an ex vivo perfusion device in cardiac transplantation introduces a unique opportunity for treating cardiac allografts with therapeutic interventions to improve function and avoid deleterious recipient responses. Establishing a translational, large-animal model for therapeutic delivery to the entire allograft is essential for testing novel therapeutic approaches in cardiac transplantation. The porcine, heterotopic heart transplantation model in the intraabdominal position serves as an excellent model for assessing the effects of novel interventions and the immunopathology of graft rejection. This model additionally offers long-term survival for the pig, given that the graft is not required to maintain the recipient's circulation. The aim of this protocol is to provide a reproducible and robust approach for achieving ex vivo delivery of a therapeutic to the entire cardiac allograft prior to transplantation and provide technical details to perform a survival heterotopic transplant of the ex vivo perfused heart.

We present a protocol for utilizing a normothermic ex vivo sanguinous perfusion system for the delivery of therapeutics to an entire cardiac allograft in a porcine heterotopic heart transplant model.

심장 동종이식편에 치료제를 전달하기 위한 돼지 이질성 심장 이식 프로토콜

Cardiac transplantation is the gold standard treatment for end-stage heart failure. However, it remains limited by the number of available donor hearts ...

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 ...