M.-A. Deutsch is supported by Dr. Rusche Forschungsprojekt (2014) Deutsche Stiftung f&#252;r Herzforschung and Deutsche Gesellschaft f&#252;r Thorax- Herz- und Gef&#228;&#223;chirurgie. M. Krane is supported by Deutsche Stiftung f&#252;r Herzforschung (F/37/11), Deutsches Zentrum f&#252;r Herz Kreislauf Forschung (DZHK B 13-050A; DZHK B 14-013SE) and Deutsche Forschungsgemeinschaft (KR3770/7-1; KR3770/9-1)

모든 실험은 독일 심장 센터 뮌헨과 바이에른, 독일 (GZ. 55.2-1-54-2532.3-17-13)의 정부에 의해 승인되었다. 동물은 특정 병원체 무료 시설에 보관하고 의학 연구를위한 국가 사회와 국립 아카데미에서 제공하는 &quot;실험 동물의 관리 및 사용을위한 가이드&quot;에 의해 공식화 &quot;실험 동물 관리의 원칙&quot;을 준수하여 인간의 치료를받은 과학 및 국립 보건원 (NIH 출판 NO. 86-23가 1985 년 개정)에 의해 출판. 참고 : 두 개의 곡선 집게 및 선박 dilatators 포함 멸균 미세 수술 장비의 완전한 세트는, 절차 (그림 2a) 필요합니다. 수술 전후 예방 적 항생제는 일상적으로 필요한 것은 아니지만 면역 동물에 표시 될 수있다. 동물 단말기 이소 플루 란 흡입을 이용하여 희생되었다. 기증자 준비 및 이식 수확 &lt;OL&gt; <li> 복강 내 (IP)에 의해 동물의 주사 미다 졸람 (5.0 ㎎ / ㎏), 메데 토미 (0.5 ㎎ / ㎏) 및 펜타닐 (0.05 ㎎ / ㎏ 체중)를 마취. 수술 분야에 부정사 위치에 마우스를 놓고 페달 철수 반사에 의해 수술 내성을 확인합니다. </li><li> 클로르헥시딘으로 피부를 세 번 소독하고 중간 선 복부 절개를 수행합니다. 하대 정맥을 노출 왼쪽에 젖은 거즈로 장을 후퇴. </li><li> 항 응고에 대한 하대 정맥을 통해 0.4 mL의 생리 식염수에 희석 헤파린의 50 IU를 주사. 헤파린 후 용기를 1 분 절개. </li><li> 다이어프램을 열고 심장을 노출하는 양자 개흉술을 수행합니다. 앞쪽에 가슴 벽 우량 반영 클램프와 장소에서 개최됩니다. </li><li> 흉선을 제거하고 퉁명스럽게 대동맥과 폐동맥 사이의 결합 조직을 해부하다. 우측 상대 정맥을 절개. </li><li> 의 수준에서 동원 상행 대동맥에 구멍30 G 바늘 팔 머리 트렁크는 5 ML의 주사기에 장착 천천히 4 ℃ 저온 심정지 용액 3 ㎖에 과도한 압력없이 retrogradually 마음을 관류. 관상 동맥에 공기의 유입을 피하십시오. 추가로 심정지를 향상 그래프트 상 빙냉 식염수 놓는다. </li><li> 팔 머리 트렁크의 기원에 근위 대동맥을 가로로 쪼개다. </li><li> 8-0 실크 심장에 가까운 열등 우수한 venae 모두 cavae을 결찰과 합자로 원심을 가로로 쪼개다. </li><li> 같은 말단 가능한 폐 트렁크를 무료 해부하다. 8-0 실크 분기에 가까운 폐 동맥을 결찰하고 원심을 가로로 쪼개다. </li><li> 단일 8-0 실크 봉합사로 모두 함께 폐 혈관을 결찰하고 그들을 가로로 쪼개다. </li><li> 기증자로부터 심장을 제거하고 이식까지 얼음처럼 차가운 심정지 용액에 보관하십시오. </li></ol> 2.받는 사람 준비 <ol><li> 준비11 호 메스 또는 현미경 가위로 폴리이 미드 튜브를 절단하여 문합을위한 커프스. 커프는 원통형 몸체 및 혈관 클램프의 배치를위한 확장 (원주의 1 / 3의), 1mm (도 2b)의 길이가 각각 이루어져있다. </li><li> 동물을 마취하고 전술 한 바와 같이 수술 내성을 확인. 유체 교체 0.25 ml의 따뜻한 식염수의 피하 (SC)를 주입한다. </li><li> 전기 면도기를 사용하여 오른쪽 자궁 지역의 머리를 제거하고 따뜻한 패드 앙와위에서 동물을 배치합니다. 호흡을 손상시킬 수있는이 같은 전면 사지를 overstretching 피하면서 테이프의 가닥 다리를 고정합니다. 건조 함을 방지하기 위해 안과 용 연고를 사용하여 눈을 커버. 수술 부위를 클로르헥시딘으로 3 회 소독한다. </li><li> 경정맥 노치에 오른쪽 하악 각도에서 가로 피부 절개를합니다. </li><li> 퉁명스럽게 내선 동원ernal 경정맥. 바이폴라 집게와 곁가지를 소작하고 그들을 가로로 쪼개다. </li><li> 턱밑 샘에서 정맥과 합류의 수준에서 합자 사이의 외부 경정맥을 나눈다. </li><li> 커프를 통해 외부 경정맥을 당겨 커프의 확장에 혈관 클램프를 배치하여 근위 혈관을 폐색. 합자를 제거 1:10 헤파린 식염수로 혈관 내강을 관개. </li><li> 커프를 통해 혈관 벽을 에버트와 사전 원주 8-0 실크 봉합사로 고정합니다. </li><li> 흉쇄 근육의 피상적 인 부분을 제거합니다. </li><li> 오른쪽 경동맥을 동원 경동맥 분기 아래 합자 사이의 혈관을 가로로 쪼개다. </li><li> 커프를 통해 경동맥을 당겨 커프의 확장에 Yasargil 클램프를 배치하여 근위 혈관을 폐색. 합자를 제거 1:10 헤파린 식염수로 혈관 내강을 관개. </li><li> 부드럽게 dilat전자 혈관 dilatator와 동맥 내강은 커프를 통해 혈관 벽을 에버트와 사전 원주 8-0 실크 봉합사로 고정합니다. </li><li> 이식 주입을위한 충분한 공간을 확보하기 위해, 턱밑 샘의 오른쪽 엽을 제거합니다. </li><li> 이식까지 촉촉한 수술 필드를 유지합니다. </li></ol> 3. 이식 주입 <ol><li> 거꾸로 내측 지향 대동맥과 측면 폐동맥과받는 사람 자궁 경부 지역에서 이식을 놓습니다. </li><li> 경동맥과 팔목에 걸쳐 이식의 대동맥을 당겨 원주 8-0 실크 봉합사로 문합을 확보. </li><li> 그 앞쪽 측면에 폐 트렁크를 절개와 외부 경정맥과 팔목에 그것을 잡아 당깁니다. 또 원주 8-0 실크 봉합사로 문합을 확보. </li><li> 경동맥의 언 클램프에 의해 다음, 외부 경정맥에 클램프를 제거합니다. 마음은 혈액 즉시 채 웁니다. R을 향상시키기eperfusion는 이식 상에 37 ° C 따뜻한 식염수를 놓습니다. 일반적으로 심장은 1 분 내에 동 율동을 개발하고 있습니다. 정맥 사이트에서 커프 확장을 제거합니다. </li><li> 올바른 이식 위치를 확인, 문합은 비틀림없이 긴장이없는 선박이어야합니다. 이 이후 피부의 소작 혈관은 상처 치유를 손상시킬 수는 없습니다. 일반적으로 출혈 자발적으로 중지합니다. 단일 8-0 연속 봉합으로 피부를 닫습니다. </li></ol> 4. 수술 후 케어 <ol><li> 유체 교체 0.25 ml의 따뜻한 식염수 사우스 캐롤라이나를 주입한다. </li><li> 마취제는 날록손 (1.2 ㎎ / ㎏), 플루 마제 닐 (0.5 ㎎ / ㎏)과는 흉골 드러 누움을 유지하기에 충분한 의식을 회복 할 때까지 따뜻한 패드 위에 동물 왼쪽 SC atipamezol (2.5mg / 체중 kg)을 주사 길항. 비정상적인 행동을 등록하는 적어도 하나의 추가 시간 동안 동물을 관찰한다. 완전히 recove 때까지 다른 동물의 회사에 수술을 한 동물을 반환하지 않습니다빨간색. </li><li> 부 프레 노르 핀과 선제 진통제를 제공 (0.05 ㎎ / ㎏ SC 세 하루에 번) 또는 시술 후 72 시간의 최소 기간 동안 지역 기관 프로토콜에 따라 다른 진통제. </li><li> 외과 의사와 수의사는 동물을 정기적으로 볼 수 있습니다. 체중 감소&gt; 15 %, 무관심, 부종 또는 외과 측 감염 동물의 경우 단말기 이소 플루 란 흡입을 사용하여 안락사된다. </li></ol>

<ol>
	<li>Corry, R. J., Winn, H. J., Russell, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+transplantation+in+congenic+strains+of+mice.">Heart transplantation in congenic strains of mice.</a> Transplant Proc. 5, 733-735 (1973).</li><li>Chen, Z. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+technique+of+cervical+heterotopic+heart+transplantation+in+mice.">A technique of cervical heterotopic heart transplantation in mice.</a> Transplantation. 52, 1099-1101 (1991).</li><li>Martins, P. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+graft+function+in+rodent+models+of+heart+transplantation.">Assessment of graft function in rodent models of heart transplantation.</a> Microsurgery. 28, 565-570 (2008).</li><li>Schramm, 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=The+subepicardial+microcirculation+in+heterotopically+transplanted+mouse+hearts:+an+intravital+multifluorescence+microscopy+study.">The subepicardial microcirculation in heterotopically transplanted mouse hearts: an intravital multifluorescence microscopy study.</a> J Thorac Cardiovasc Surg. 134, 210-217 (2007).</li><li>Liu, F., Kang, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterotopic+heart+transplantation+in+mice.">Heterotopic heart transplantation in mice.</a> J Vis Exp : JoVE. 238, (2007).</li><li>Gong, 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=Introduction+of+modified+cervical+cardiac+transplant+model+in+mice.">Introduction of modified cervical cardiac transplant model in mice.</a> Exp Clin Transplant. 10, 158-162 (2012).</li><li>Matsuura, A., Abe, T., Yasuura, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simplified+mouse+cervical+heart+transplantation+using+a+cuff+technique.">Simplified mouse cervical heart transplantation using a cuff technique.</a> Transplantation. 51, 896-898 (1991).</li><li>Chen, H., Zhang, Y., Zheng, D., Praseedom, R. K., Dong, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+kidney+transplantation+in+mice:+technique+using+cuff+for+renal+vein+anastomosis.">Orthotopic kidney transplantation in mice: technique using cuff for renal vein anastomosis.</a> PloS one. 8, e77278 (2013).</li><li>Maglione, M., 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=A+novel+technique+for+heterotopic+vascularized+pancreas+transplantation+in+mice+to+assess+ischemia+reperfusion+injury+and+graft+pancreatitis.">A novel technique for heterotopic vascularized pancreas transplantation in mice to assess ischemia reperfusion injury and graft pancreatitis.</a> Surgery. 141, 682-689 (2007).</li><li>Sucher, 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=Orthotopic+hind-limb+transplantation+in+rats.">Orthotopic hind-limb transplantation in rats.</a> J Vis Exp : JoVE. , (2010).</li><li>Kamada, N., Calne, R. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+the+rat.+Technique+using+cuff+for+portal+vein+anastomosis+and+biliary+drainage.">Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage.</a> Transplantation. 28, 47-50 (1979).</li><li>Oldani, G., Lacotte, S., Morel, P., Mentha, G., Toso, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+rats.">Orthotopic liver transplantation in rats.</a> J Vis Exp : JoVE. , (2012).</li><li>Okazaki, M., 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=A+mouse+model+of+orthotopic+vascularized+aerated+lung+transplantation.">A mouse model of orthotopic vascularized aerated lung transplantation.</a> Am J Transplant. 7, 1672-1679 (2007).</li><li>Suzuki, H., Fan, L., Wilkes, 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=Development+of+obliterative+bronchiolitis+in+a+murine+model+of+orthotopic+lung+transplantation.">Development of obliterative bronchiolitis in a murine model of orthotopic lung transplantation.</a> J Vis Exp : JoVE. , (2012).</li><li>Zou, Y., 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=Mouse+model+of+venous+bypass+graft+arteriosclerosis.">Mouse model of venous bypass graft arteriosclerosis.</a> Am J Pathol. 153, 1301-1310 (1998).</li><li>Zhou, Y., Gu, X., Xiang, J., Qian, S., Chen, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparative+study+on+suture+versus+cuff+anastomosis+in+mouse+cervical+cardiac+transplant.">A comparative study on suture versus cuff anastomosis in mouse cervical cardiac transplant.</a> Exp Clin Transplant. 8, 245-249 (2010).</li><li>Niimi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+technique+for+heterotopic+cardiac+transplantation+in+mice:+experience+of+3000+operations+by+one+surgeon.">The technique for heterotopic cardiac transplantation in mice: experience of 3000 operations by one surgeon.</a> J Heart Lung Transplant. 20, 1123-1128 (2001).</li><li>Arras, M., Autenried, P., Rettich, A., Spaeni, D., Rulicke, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+of+intraperitoneal+injection+anesthesia+in+mice:+drugs,+dosages,+adverse+effects,+and+anesthesia+depth.">Optimization of intraperitoneal injection anesthesia in mice: drugs, dosages, adverse effects, and anesthesia depth.</a> Comp Med. 51, 443-456 (2001).</li></ol>

The authors have nothing to disclose.

우리는 다양한 실험 설정에서 90 % 이상의 성공률과 200 개 이상의 심장 이식을 완료했습니다. 여기에 설명 된 기술로, 전체 절차는 숙련 된 의사에 의해 60 분 내에 완료 될 수있다. 장기 조달 전에 자궁 혈관을 제조함으로써, 총 그래프트 허혈 시간을 20 분으로 제한 할 수있다. 주입 (온 허혈 시간) 대 시간 봉합술 (16)에 비해 커프 기술을 이용하여 현저하게 짧다. 대신 ?...

심장 이식 모델은 자주 I / R 상해, 동종 이식 거부 반응, 이식 후 혈관 및 면역 에이전트의 효율성을 조사하는 데 사용됩니다. 마우스 모델은 공지 된 면역 학적 및 유전 적 배경이 많은 근친 형질 전환 균주 및 상대적으로 낮은 비용의 실험 가용성과 같은 많은 이점을 결합한다. 1973 년 생쥐의 복부 심장 이식하는 기술이 먼저 코리 등 1에 의해 설명되었다. 이 모델에서 이식받는 사람의 복부 대동맥과 하대 정맥에 주 폐동맥의 상행 대동맥의 엔드 - 투 - 측 문합에 의해 revascularized된다. 1991 년 생쥐의 자궁 심장 이식 봉합 기술은 첸 등의 알 2에 의해 설명되었다. 이 모델에서 역행 관상 동맥의 관류는 GR의 상행 대동맥에받는 사람의 경동맥 동맥의 문합에 의해 설립선미. 정맥 혈액 우심방, 우심실에 관상 정맥동을 통해 배수 및받는 사람의 외부 경정맥 정맥 (그림 1)에 폐동맥을 통해 배출됩니다. 복부 이식 주입에 비해 자궁 모델은 덜 침습적이며, 이식의 표면 위치는 추가로 추적 검사 (예 촉진, 심장 초음파 검사, 생체 내에 형광 현미경) 3,4를위한 아주 쉽게 접근 할 수 있습니다. 기술 향상 5,6에도 불구하고,이 모델의보다 광범위한 사용은 문 합부 누출과 혈전증의 발생률이 높은과 작은 혈관의 봉합 문합을 수행하기 때문에 기술적 인 문제에 국한되고있다. 1991 년, 마츠우라 등. 혈관벽 크게 자궁 심장 이식 7의 모델을 용이하게 합성 실린더 위에 everted되는 커프 기술을 소개했다. 이 기술은 푸이었다rther 신장 (8), 췌장 (9), 사지 (10), 간 11, 12, 13, 14 폐 및 혈관 이식 (15) 등 다양한 실험 이식 모델에 채택했다. 봉합 기술에 비해 필요한 문합을위한 시간과, 따라서, 온 허혈 손상 시간이 짧고에 커프 (16)의 기술을 이용하여 상당히 적은 혈관 합병증이있다. 또한, 높은 성공률로 동작을 수행하도록 미세 작은 외과 훈련 할 수있다. 커프 기술의 단점은 대뇌 재관류를 변경할 수 경동맥의 결찰 필수이다. 이 비디오 논문에서는 생쥐의 자궁 심장 이식을위한 개선 및 단순화 된 커프 기술을 제시한다.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Forceps JFC-7.2</td>
			<td>S&#38;T</td>
			<td>P-00036</td>
			<td>Curved tip 0.20 mm</td>
		</tr>
		<tr>
			<td>Vessel dilatator D-5a</td>
			<td>S&#38;T</td>
			<td>S-00124</td>
			<td></td>
		</tr>
		<tr>
			<td>Adventita scissor</td>
			<td>S&#38;T</td>
			<td>S-00102</td>
			<td></td>
		</tr>
		<tr>
			<td>Vascular clamp B-1V</td>
			<td>S&#38;T</td>
			<td>S-00396</td>
			<td></td>
		</tr>
		<tr>
			<td>Yasargil Clip</td>
			<td>Peter Lazic</td>
			<td>65,097</td>
			<td>3.5 mm lenght</td>
		</tr>
		<tr>
			<td>Bipolar forceps</td>
			<td>Micromed</td>
			<td>148-100-011</td>
			<td>tip 0.25 mm</td>
		</tr>
		<tr>
			<td>Polyimide tubes</td>
			<td>River Tech Medical</td>
			<td></td>
			<td>19-24 gauge</td>
		</tr>
		<tr>
			<td>8.0 silk ligatures</td>
			<td>Catgut</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Custodiol HTK solution</td>
			<td>Essential Pharmaceuticals</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

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.

우리 팀은 성공적으로 약 20 훈련 수술 후 설명 된 기술을 수행 할 수 있습니다. 가장 어려운 부분은 쉽게 다시 플립 여러 시도가 찢어 벽을 초래할 수 있기 때문에, 커프스 위에 경동맥 혈관 벽을 에버트한다. 훈련 기간 후에, 평균 총 수술 시간은 60 ± 8 분이었다. 약 20 분은 기증자 수확, 자궁 혈관 문합과 피부 폐쇄에 대한 15 분 준비를위한 25 분 필요합니다. 심근 I / R 상해의 ...

Watch this Scientific Journal Video about Heterotopic Cervical Heart Transplantation in Mice at JoVE.com

Heterotopic Cervical Heart Transplantation in Mice

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 ...

heterotopic-cervical-heart-transplantation-in-mice

Research

JoVE Journal

Medicine

11.0K 조회수.  German Heart Center Munich, Technische Universität München. 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.

비디오: Heterotopic Cervical Heart Transplantation in Mice

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

마우스의 제어 자궁 경부 열상 상해

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

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 ...

A Modified Method for Heterotopic Mouse Heart Transplantion

Mice are often used as heart transplant donors and recipients in studies of transplant immunology due to the wide range of transgenic mice and reagents available. A difficulty is presented due to the small size of the animal and the considerable technical challenges of the microsurgery involved in heart transplantation. In particular, a high rate of technical failure early after transplantation may result from recipient death and post-operative complications such as hind limb paralysis or a non-beating heart. Here, the complete technique for heterotopic mouse heart transplantation is demonstrated, involving harvesting the donor heart and its subsequent implantation into a recipient mouse. The donor heart is harvested immediately following in situ perfusion with cold heparinized saline and transection of the ascending aorta and pulmonary artery. The recipient operation involves preparation of the abdominal aorta and inferior vena cava (IVC), followed by end-to-side anastomosis of the donor aorta with the recipient aorta using a single running 10-0 microsuture and a similar anastomosis of the donor pulmonary artery with the recipient IVC. Following the operation the animal is injected with 0.6 ml normal saline subcutaneously and allowed to recover on a 37&#160;&#176;C heating pad. The results from 227 mouse heart transplants are summarized with a success rate at 48 hr of 86.8%. Of the 13.2% failures within 48 hr, 5 (2.2%) experienced hind limb paralysis, 10 (4.4%) had a non-beating heart due to graft ischemic injury and/or thrombosis, while 15 (6.6%) died within 48 hr.

A technique is demonstrated for the microsurgical procedure for heterotopic transplantation of hearts in mice, including simplified methods for donor harvesting and recipient vessel anastomosis.

이소성 마우스 심장 장기 이식에 대한 수정 방법

Mice are often used as heart transplant donors and recipients in studies of transplant immunology due to the wide range of transgenic mice and reagents ...

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 ...

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

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed results. However, the cellular and molecular mechanisms of the neotissue development, valve thickening, and stenosis development are not researched extensively. To answer the above questions, we developed a murine heterotopic heart valve transplantation model. A heart valve was harvested from a valve donor mouse and transplanted to a heart donor mouse. The heart with a new valve was transplanted heterotopically to a recipient mouse. The transplanted heart showed its own heartbeat, independent of the recipient&#8217;s heartbeat. The blood flow was quantified using a high frequency ultrasound system with a pulsed wave Doppler. The flow through the implanted pulmonary valve showed forward flow with minimal regurgitation and the peak flow was close to 100 mm/sec. This murine model of heart valve transplantation is highly versatile, so it can be modified and adapted to provide different hemodynamic environments and/or can be used with various transgenic mice to study neotissue development in a tissue engineered heart valve.

In order to understand the cellular and molecular mechanisms underlying neotissue formation and stenosis development in tissue engineered heart valves, a murine model of heterotopic heart valve transplantation was developed. A pulmonary heart valve was transplanted to recipient using the heterotopic heart transplantation technique.

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 ...

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 ...

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 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 ...