Name: mouse pneumonectomy model of compensatory lung growth
Uploaded: 2014-12-17T14:00:00.000+00:00
Duration: 9 min 22 s
Description: Watch this Scientific Journal Video about Mouse Pneumonectomy Model of Compensatory Lung Growth at JoVE.com

The authors would like to acknowledge the Cincinnati Children&#8217;s Hospital Division of Veterinary Services for their assistance. This manuscript was supported by the National Institutes of Health K12 HD028827. Anna Perl PhD taught the authors this surgical procedure.

참고 : 동물 use 문 : 본 연구의 모든 절차를 승인 신시내티 아동 병원의 기관 동물 사용 및 관리위원회 (IACUC)의 가이드 라인을 다음과 같이 실시 하였다. 여덟 주령의 C57BL / 6J 수컷 마우스는 잭슨 연구소 (주 바 하버, ME)로부터 획득하고 사용하기 전에 일주일 동안 순응시켰다. 수술까지는 동물은 무균 장벽 시설에 보관하고 멸균 차우 여과 물 광고 libdium를 제공했다. 각 마우스 케이지는 전용 공기와 물을 공급하고, 객실은 12 시간 밤낮주기를 유지 하였다. 수술 회복 후, 마우스, 여과 탑과에서 사육 멸균 우 광고 섭취하는을 제공하고, 물병에서 물을 여과 제공되었다. 악기 1. 준비 <ol><li> 종이 클립과 핀을 사용하여 6 피부 견인기를 확인합니다. 트위스트 종이 핀의 정강이에 종이 클립을 곧게, L일단에 5cm 직선 강선 처마와 와이어의 단부에 단일 0.5 cm &quot;U&quot;자 고리를 만든다. </li><li> 플라스틱 랩을 사용하여 약 15 X 15cm 정사각형 외과 드레이프를 확인합니다. 마우스 당 하나의 드레싱을 준비합니다. 각 랩 사이에 종이 타워를 넣습니다. </li><li> 12 × 12 인치 코르크 타일, 거즈, 그리고 면봉 면봉의 스택과 함께 모든 수술 도구를 소독. </li></ol> 2. 마우스 준비 <ol><li> 2 % 이소 플루 란 마취를 유도한다. 동물의 무게를. </li><li> 전용 수술 준비 영역에서 전기 면도기 왼쪽 흉부와 목 영역을 면도. </li><li> 마우스의 눈에 인공 눈물 연고의 드롭을 적용합니다. </li><li> 클로르헥시딘과 이소 프로필 알코올 목과 왼쪽 가슴의 오염을 제거. 두 번 더 반복한다. </li></ol> 3. 마우스 오로 - 기관 내 삽관 및 기계 환기 <ol><li> 비 멸균 수술 기술자 장소를 예열 외과 영역에서 마우스 부정사가있다.</li><li> 발 핀치에 대한 응답의 부족을 문서화하여 마취의 깊이를 확인합니다. </li><li> 수술 복장, 마스크, 모자를 손을 씻고 걸치고 후, 멸균 수술 장갑을 돈. </li><li> 입체 재단 및 무균 기술을 사용하면, 후두를 노출 앞쪽 중간 목을 통해 1cm 세로 절개를합니다. 살짝 곡선, 톱니 모양의 10cm 집게와 스트랩 근육을 철회하고 직선 가위의 끝 스트랩 근육을 확산하여 후두와 기관을 노출. </li><li> 경구 중반 기관 (그림 1A)에 22 G 뭉툭한 팁 angiocatheter를 삽입하고 시각적으로 배치 (그림 1B)를 확인합니다. 마취를 유지하고 설치류 인공 호흡기를 통해 (스트로크 당 225 μL, 분 당 200 스톡스를) 1-3% 이소 플루 란 (isoflurane)을 이용 환기. 15cm의 H 2 O의 압력 한계를 채용 </li></ol> 4. 마우스 전폐 절제술 <ol><li> 마우스의 뒷면을 향 우측 측와위 위치에 마우스를 놓는다연산자 (왼쪽 위). 멸균 드레이프로 자체 밀봉 플라스틱 랩을 사용합니다. 드레이프를 통해 절단, 4 번째와 5 번째 늑간에서 갈비뼈에 2cm 길이 컷 평행하게 끝이 무딘 곡선 가위를 사용합니다. 무딘 끝 곡선 가위를 삽입하고 기본 갈비와 늑간 근육에서 멀리 피부를 해부하다. </li><li> 광장을 1.5 × 1.5 cm 수술 창 (그림 2A)를 노출 네 견인기로 피부를 후퇴. 코르크 보드에 견인기를 고정합니다. </li><li> 곡선 집게를 사용 갈비뼈 아래로 해부하고, 흉강을 입력 곡선 집게 중 하나 팁을 사용합니다. </li><li> 뭉툭한 팁 마이크로 가위를 사용하여, 흉강을 입력 하부 날개를 사용합니다. 반대 방향으로 갈비와 반복 사이에 0.5 cm의 절개를합니다. </li><li> 나머지 두 견인기를 사용하여 전후방 축과 코르크 보드 (그림 2B)로 견인기 보안에 흉부를 엽니 다. </li> &lt;왼쪽 폐 동맥과 기관지가 (그림 3A, B)에 노출 될 때까지 왼쪽 손에 곡선 무딘 팁 집게를 사용하여 리&gt;, 개흉술을 통해 옆으로 하방 왼쪽 폐의 상부를 왼쪽 폐를 잡고 대체. </li><li> 손바닥에 도포 손목 (그림 3C)에서 멀리 가리키는 곡선 팁의 시체와 함께 오른쪽에있는로드 티타늄 혈관 microclip 주걱을 잡고, 왼쪽의 후방 측면의 곡률을 따라 흉부에 도포 팁을 밀어 폐는 왼쪽 기관지 및 폐 동맥 (그림 3D)를 클립. </li><li> 도포를 제거하지만 수축 왼쪽 폐를 유지합니다. 오른손으로 무딘 팁 마이크로 가위를 잡고 클립에 기관지 및 폐 동맥 원위부 절단과 왼쪽 폐 (그림 3E)를 제거합니다. </li><li> 리브 견인기를 제거합니다. </li><li> incisi 열등 피부의 1cm를 끼지 곡선 무딘 집게를 사용하여에하지만 진동판의 수준 이상과 피부를 통해 왼쪽 흉강 (그림 4A, B)에 24 G의 angiocatheter를 삽입합니다. </li><li> 흉강을 닫습니다 제 4 회 약 2 단속 봉합 5 일 리브를 배치 5-0 prolene의 봉합을 사용합니다. </li><li> 피부 견인기를 제거합니다. 절개의 길이를 따라 피부를 피부에 근사하고 폐쇄 붙 집게 두 세트를 사용한다. </li><li> angiocatheter에 3 ㎖ 루어 잠금 주사기를 연결하고 부드러운 흡입을 적용하고 angiocatheter을 인출하여 잔류 공기를 제거합니다. </li><li> 목 절개 전과 집게 두 세트를 사용하는 폐쇄 붙인다. </li></ol> 5. 마우스 위장 전폐 절제술 <ol><li> &quot;마우스 전폐 절제술&quot;프로토콜에서 언급 한 바와 같이 왼쪽 폐를 노출. 왼쪽 흉강에 공기를 허용하는 곡선 무딘 집게로 갈비뼈를 들어 올립니다 (그림 5A, B). </li><li> 왼쪽 상점에 24 G의 angiocatheter를 배치왼쪽 폐를 손상하지 않도록주의 상기와 흉부 캐비티. </li><li> 5-0 prolene의 봉합을 사용하고 폐 (그림 5C)를 구멍을 뚫거나하지 않도록주의, 제 3 회 / 4 회에 봉합사의 두 길이와 5 일 / 6 일 리브의 사이 공간 (그림 5D)를 배치합니다. 왼쪽 폐 탈출증의 위험을 줄이기 위해 묶는 전에 봉합사의 양쪽 길이를 놓습니다. 두 중단 바늘 (그림 5E)을 만들 수있는 봉합사를 묶어. </li><li> 흉부 절개를 통해 피부 접착제, angiocatheter에 잔류 공기를 제거하고, 상기와 목 절개를 붙입니다. </li></ol> 6. 소생술, 진통, 및 복구 <ol><li> 이소 플루 란의 전원을 끄고, 부 프레 노르 핀의 0.1 ㎎ / ㎏을 관리하고 생리 식염수를 피하 0.5 ml의. </li><li> 자발 호흡이 다시 시작하면, 기관 튜브를 제거합니다. </li><li> 다시 외래 때까지 마우스를 관찰합니다. 일반적으로 resum 워킹기관 내 튜브의 제거 후 몇 분 말이지. </li><li> / N을 O를 복구하기 위해 27 ° C 배양기 (가습, 25 % 산소)에 마우스를 놓습니다. 참고 : 우리는 수술 후 첫 24 시간 동안 케이지 바닥에 물을 적신 차우의 몇 가지 알약을 배치합니다. </li><li> 수술 후 하루에 두 번 3 일 동안 복강 주사로 부 프레 노르 핀의 0.1 ㎎ / ㎏을 관리합니다. 동물을 처리 할 때 수술 부위를 열 않도록주의하십시오. </li></ol> 7. 마우스 모니터링 <ol><li> 수술 후 1, 3, 5 및 7 일째에 마우스 체중. </li></ol>

<ol>
	<li>Strueby, L., Thebaud, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+bronchopulmonary+dysplasia.">Advances in bronchopulmonary dysplasia.</a> Expert review of respiratory medicine. , (2014).</li><li>Donn, S. M., Sinha, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+advances+in+the+understanding+and+management+of+bronchopulmonary+dysplasia.">Recent advances in the understanding and management of bronchopulmonary dysplasia.</a> Seminars in fetal &amp; neonatal medicine. 14, 332 (2009).</li><li>Molen, T., Miravitlles, M., Kocks, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=COPD+management:+role+of+symptom+assessment+in+routine+clinical+practice.">COPD management: role of symptom assessment in routine clinical practice.</a> International journal of chronic obstructive pulmonary disease. 8, 461-471 (2013).</li><li>Seifert, A. 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=Skin+shedding+and+tissue+regeneration+in+African+spiny+mice+(Acomys).">Skin shedding and tissue regeneration in African spiny mice (Acomys).</a> Nature. 489, 561-565 (2012).</li><li>Vidal, P., Dickson, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regeneration+of+the+distal+phalanx.+A+case+report.">Regeneration of the distal phalanx. A case report.</a> Journal of hand surgery. 18, 230-233 (1993).</li><li>Potter, P. C., Levine, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bone+Regeneration+Following+Chronic+Suppurative+Osteitis+of+the+Distal+Phalanx.">Bone Regeneration Following Chronic Suppurative Osteitis of the Distal Phalanx.</a> Annals of surgery. 80, 728-729 (1924).</li><li>McKim, L. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regeneration+of+the+Distal+Phalanx.">Regeneration of the Distal Phalanx.</a> Canadian Medical Association journal. 26, 549-550 (1932).</li><li>Brown, L. M., Rannels, S. R., Rannels, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implications+of+post-pneumonectomy+compensatory+lung+growth+in+pulmonary+physiology+and+disease.">Implications of post-pneumonectomy compensatory lung growth in pulmonary physiology and disease.</a> Respir Res. 2, 340-347 (2001).</li><li>Holder, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regeneration+and+compensatory+growth.">Regeneration and compensatory growth.</a> British medical bulletin. 37, 227-232 (1981).</li><li>Hsia, C. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lessons+from+a+canine+model+of+compensatory+lung+growth.">Lessons from a canine model of compensatory lung growth.</a> Curr Top Dev Biol. 64, 17-32 (2004).</li><li>Butler, J. P., 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=Evidence+for+adult+lung+growth+in+humans.">Evidence for adult lung growth in humans.</a> N Engl J Med. 367, 244-247 (2012).</li><li>Konerding, M. 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=Spatial+dependence+of+alveolar+angiogenesis+in+post-pneumonectomy+lung+growth.">Spatial dependence of alveolar angiogenesis in post-pneumonectomy lung growth.</a> Angiogenesis. 15, 23-32 (2012).</li><li>Kho, A. T., Liu, K., Visner, G., Martin, T., Boudreault, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+dedifferentiation+and+redevelopment+phases+during+postpneumonectomy+lung+growth.">Identification of dedifferentiation and redevelopment phases during postpneumonectomy lung growth.</a> Am J Physiol Lung Cell Mol Physiol. 305, 542-554 (2013).</li><li>Wang, W., Nguyen, N. M., Guo, J., Longitudinal Woods, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noninvasive+Monitoring+of+Compensatory+Lung+Growth+in+Mice+after+Pneumonectomy+via+(3)He+and+(1)H+Magnetic+Resonance+Imaging.">Noninvasive Monitoring of Compensatory Lung Growth in Mice after Pneumonectomy via (3)He and (1)H Magnetic Resonance Imaging.</a> Am J Respir Cell Mol Biol. 49, 697-703 (2013).</li><li>Gibney, B. C., 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=Detection+of+murine+post-pneumonectomy+lung+regeneration+by+18FDG+PET+imaging.">Detection of murine post-pneumonectomy lung regeneration by 18FDG PET imaging.</a> EJNMMI research. 2, 48 (2012).</li><li>Rawlins, E. L., Perl, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+a'MAZE'ing+world+of+lung-specific+transgenic+mice.">The a'MAZE'ing world of lung-specific transgenic mice.</a> Am J Respir Cell Mol Biol. 46, 269-282 (2012).</li><li>Ochs, M., Muhlfeld, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+microscopy+of+the+lung:+a+problem-based+approach.+Part+1:+basic+principles+of+lung+stereology.">Quantitative microscopy of the lung: a problem-based approach. Part 1: basic principles of lung stereology.</a> Am J Physiol Lung Cell Mol Physiol. 305, L15-22 (2013).</li><li>Ysasi, A. B., 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=Effect+of+unilateral+diaphragmatic+paralysis+on+postpneumonectomy+lung+growth.">Effect of unilateral diaphragmatic paralysis on postpneumonectomy lung growth.</a> Am J Physiol Lung Cell Mol Physiol. 305, L439-445 (2013).</li><li>Dane, D. M., Yilmaz, C., Estrera, A. S., Hsia, C. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Separating+in+vivo+mechanical+stimuli+for+postpneumonectomy+compensation:+physiological+assessment.">Separating in vivo mechanical stimuli for postpneumonectomy compensation: physiological assessment.</a> Journal of applied physiology. 114, 99-106 (2013).</li><li>Mortola, J. P., Magnante, D., Saetta, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expiratory+pattern+of+newborn+mammals.">Expiratory pattern of newborn mammals.</a> Journal of applied physiology. 58, 528-533 (1985).</li><li>MacDonald, K. D., Chang, H. Y., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+simple+method+of+mouse+lung+intubation.">An improved simple method of mouse lung intubation.</a> Journal of applied physiology. 106, 984-987 (2009).</li></ol>

The authors have nothing to disclose.

우리는 지금까지보고 된 마우스 PNX와 마우스 가짜 PNX 절차의 가장 상세한 설명을 제공하고 있습니다. 우리는 절차를 학습 연구자들은 일반적으로 발생하는 일반적인 함정의 여러 가지의 독자가 인식 한 우리는이 함정을 완화하기 위해 실험실에서 개발 한 몇 가지 기술을 설명했다. 이 모델을 이용하여 다른 실험실은 다른 기술 수​​정을 개발하거나 다른 악기를 사용할 수 있습니다. 기술의 차?...

폐의 주요 기능은 유기체와 대기 사이의 산소와 이산화탄소의 교환을 제공하는 것이다. 인간에서, 선천성 및 획득 조건 호스트 손상된 폐 기능의 감소 결과 폐 표면적 이어질. 즉, 폐 - 같은 흡입 코르티코 스테로이드, 기관지 확장제, 산소 보충, 만성 기계 호흡으로 치료의 호스트가 손상된 폐 기능 1-3의 결과를 완화하기 위해 사용되지만, 이러한 조건을위한 이상적인 치료는 기능 폐 조직의 재성장을 촉진 것 재생. 포유류 조직 재생이 잘 설명되어 있습니다. 아프리카 가시 마우스 반흔 4없이 피부의 넓은 지역을 다시 생성 할 수 있습니다. 인간의 말단 지골 부상 또는 절단 5-7 다음 다시 생성 할 수 있습니다. 다음 전폐 절제술 (PNX), 보상 폐 성장 할, 9 쥐, 마우스 (8) 발생GS (10), 및 11 인간. 정의에 따라 보상 폐 성장은 관련 미세 (12)의 확장과이 확대 airspaces의 기존 airspaces의뿐만 아니라 확장,하지만 다시 격막을 포함한다. 유전자 발현 분석은이 모델은 폐 (13)의 개발 시그널링 많은 이벤트를 되풀이 것을 보여 주었다. 네 주 마우스 PNX 후, 폐포 표면적은 가짜 운영 동물 (14)의 그것과 동일하다. 이 논문에서 우리는 마우스 PNX와 가짜 PNX 절차를 설명합니다.

<table><tbody><tr><td>6 inch Vascular clip applicator</td><td>Teleflex Medical (WECK)</td><td>137062</td><td></td></tr><tr><td>Horizon small titanium red clip</td><td>Teleflex Medical (WECK)</td><td>1201</td><td></td></tr><tr><td>Narrow pattern 12 cm curved forceps</td><td>Fine Science Tools</td><td>11003-12</td><td></td></tr><tr><td>Curved serrated 10 cm Graefe forceps</td><td>Fine Science Tools</td><td>11052-10</td><td></td></tr><tr><td>Castroviejo needle holder</td><td>Fine Science Tools</td><td>12565-14</td><td></td></tr><tr><td>Straight 9 cm Strabismus scissors (blunt tip)</td><td>Fine Science Tools</td><td>14075-09</td><td></td></tr><tr><td>Straight 8.5 cm hardened fine scissors</td><td>Fine Science Tools</td><td>14090-09</td><td></td></tr><tr><td>Straight, blunt tip Cohan-Vannas spring scissors</td><td>Fine Science Tools</td><td>15000-12</td><td></td></tr><tr><td>Skin glue</td><td>Gluture</td><td>32046</td><td></td></tr><tr><td>22 G Angiocatheter</td><td></td><td></td><td></td></tr><tr><td>24 G Angiocatheter</td><td></td><td></td><td></td></tr><tr><td>3 ml Luer lock syringe</td><td></td><td></td><td></td></tr><tr><td>4 Short retractors</td><td></td><td></td><td></td></tr><tr><td>2 Long retractors</td><td></td><td></td><td></td></tr><tr><td>5-0 Prolene on curved cutting needle</td><td>Ethicon</td><td>8698G</td><td></td></tr><tr><td>0.5 ml Syringe on 27 G needle</td><td></td><td></td><td></td></tr><tr><td>Normal saline</td><td></td><td></td><td></td></tr><tr><td>Buprenorphine</td><td></td><td></td><td></td></tr><tr><td>Press-n-Seal wrap</td><td>Glad Products Company</td><td></td><td></td></tr><tr><td>12 x 12 inch Cork board stack</td><td>Office Depot</td><td></td><td></td></tr><tr><td>70% Ethanol</td><td></td><td></td><td></td></tr><tr><td>Betadine</td><td></td><td></td><td></td></tr><tr><td>Mouse ventilator</td><td>Hugo Sachs Elektronnik</td><td>Minivent Type 845</td><td></td></tr><tr><td>Isoflurane vaporizer</td><td>OHMEDA</td><td>Excel 210 SE</td><td></td></tr><tr><td>Artificial tear ointment</td><td>Puralube</td><td>NDC: 17033-211-38</td><td></td></tr></tbody></table>

mouse pneumonectomy model of compensatory lung growth

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to disability or death. Injury-based animal models of lung repair and regeneration are limited by injury-specific responses making it difficult to differentiate changes related to the injury response and injury resolution from changes related to lung repair and lung regeneration. However, use of animal models to identify these repair and regeneration signaling pathways is critical to the development of new therapies aimed at improving pulmonary function following lung injury. The mouse pneumonectomy model utilizes compensatory lung growth to isolate those repair and regeneration signals in order to more clearly define mechanisms of alveolar re-septation. Here, we describe our technique for performing mouse pneumonectomy and sham pneumonectomy. This technique may be utilized in conjunction with lineage tracing or other transgenic mouse models to define molecular and cellular mechanism of lung repair and regeneration.

. 모두 PNX와 가짜 전폐 절제술 100 % - PNX와 가짜 운영 마우스 무게의 플롯은 그림 6에 제공됩니다 우리의 손에, 생존은 지속적으로 95이다. 오른쪽 폐 - 성장 다시이 모델과 예상 시간 과정에서, 우리는 기 브니 등. (15)과 왕 등의 원고에 독자를 참조하는 방법에 대한 설명. (14) 몇 가지 일반적인 함정이 성공적으로 마우스 PNX와 마...

Watch this Scientific Journal Video about Mouse Pneumonectomy Model of Compensatory Lung Growth at JoVE.com

Mouse Pneumonectomy Model of Compensatory Lung Growth

Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.

보상 성 폐 성장의 마우스 전폐 절제술 모델

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to ...

mouse-pneumonectomy-model-of-compensatory-lung-growth

Research

JoVE Journal

Medicine

12.8K 조회수.  Cincinnati Children's Hospital Medical Center. Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.

비디오: Mouse Pneumonectomy Model of Compensatory Lung Growth

Development of Obliterative Bronchiolitis in a Murine Model of Orthotopic Lung Transplantation

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and standardized not only for the study of allo-rejection but also between syngeneic strains for examining mechanisms of ischemia-reperfusion injury after lung transplantation. Although the application of the rat and other large animal model 2 contributed significantly to the elucidation of these studies, the scope of those investigations is limited by the scarcity of knockout and transgenic rats. Due to no effective therapies for obliterative bronchiolitis, the leading cause of death in lung transplant patients, there has been an intensive search for pre-clinical models that replicate obliterative bronchiolitis. The tracheal allograft model is the most widely used and may reproduce some of the histopathologic features of obliterative bronchiolitis 3. However, the lack of an intact vasculature with no connection to the recipient's conducting airways, and incomplete pathologic features of obliterative bronchiolitis limit the utility of this model 4. Unlike transplantation of other solid organs, vascularized mouse lung transplants have only recently been reported by Okazaki and colleagues for the first time in 2007 5. Applying the basic principles of the rat lung transplant, our lab initiated the obliterative bronchiolitis model using minor histoincompatible antigen murine orthotopic single-left lung transplants which allows the further study of obliterative bronchiolitis immunopathogenesis6.

Obliterative bronchiolitis is the key impediment to the long-term survival of lung transplant recipients and the lack of a robust preclinical model precludes examining obliterative bronchiolitis immunopathogenesis. Unlike other solid organ transplants, vascularized mouse lung transplantation has only recently been developed. Here we show our independently developed obliterative bronchiolitis model after murine orthotopic single-lung transplantation.

Orthotopic 폐 이식의 Murine 모델에 Obliterative Bronchiolitis의 개발

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and ...

연구

의학

A Standardized Method for Measuring Internal Lung Surface Area via Mouse Pneumonectomy and Prosthesis Implantation

Pulmonary morphology, physiology, and respiratory functions change in both physiological and pathological conditions. Internal lung surface area (ISA), representing the gas-exchange capacity of the lung, is a critical criterion to assess respiratory function. However, observer bias can significantly influence measured values for lung morphological parameters. The protocol that we describe here minimizes variations during measurements of two morphological parameters used for ISA calculation: internal lung volume (ILV) and mean linear intercept (MLI). Using ISA as a morphometric and functional parameter to determine the outcome of alveolar regeneration in both pneumonectomy (PNX) and prosthesis implantation mouse models, we found that the increased ISA following PNX treatment was significantly blocked by implantation of a prosthesis into the thoracic cavity1. The ability to accurately quantify ISA is not only expected to improve the reliability and reproducibility of lung function studies in injured-induced alveolar regeneration models, but also to promote mechanistic discoveries of multiple pulmonary diseases.

Internal lung surface area (ISA) is a critical criterion for assessing lung morphology and physiology in lung diseases and injury-induced alveolar regeneration. We describe here a standardized method that can minimize the measurement bias for ISA in both lung pneumonectomy and prosthesis implantation mouse models.

마우스 폐 전 절제술 및 보철물 삽입을 통한 내시경 표면 측정을위한 표준화 된 방법

Pulmonary morphology, physiology, and respiratory functions change in both physiological and pathological conditions. Internal lung surface area (ISA), ...

발생학

Measurement of the Pressure-volume Curve in Mouse Lungs

In recent decades the mouse has become the primary animal model of a variety of lung diseases. In models of emphysema or fibrosis, the essential phenotypic changes are best assessed by measurement of the changes in lung elasticity. To best understand specific mechanisms underlying such pathologies in mice, it is essential to make functional measurements that can reflect the developing pathology. Although there are many ways to measure elasticity, the classical method is that of the total lung pressure-volume (PV) curve done over the whole range of lung volumes. This measurement has been made on adult lungs from nearly all mammalian species dating back almost 100 years, and such PV curves also played a major role in the discovery and understanding of the function of pulmonary surfactant in fetal lung development. Unfortunately, such total PV curves have not been widely reported in the mouse, despite the fact that they can provide useful information on the macroscopic effects of structural changes in the lung. Although partial PV curves measuring just the changes in lung volume are sometimes reported, without a measure of absolute volume, the nonlinear nature of the total PV curve makes these partial ones very difficult to interpret. In the present study, we describe a standardized way to measure the total PV curve. We have then tested the ability of these curves to detect changes in mouse lung structure in two common lung pathologies, emphysema and fibrosis. Results showed significant changes in several variables consistent with expected structural changes with these pathologies. This measurement of the lung PV curve in mice thus provides a straightforward means to monitor the progression of the pathophysiologic changes over time and the potential effect of therapeutic procedures.

Here we present a protocol to simply and reliably measure the lung pressure-volume curve in mice, showing that it is sufficiently sensitive to detect phenotypic parenchymal changes in two common lung pathologies, pulmonary fibrosis and emphysema. This metric provides a means to quantify the lung&#8217;s structural changes with developing pathology.

마우스 폐의 압력 볼륨 곡선의 측정

In recent decades the mouse has become the primary animal model of a variety of lung diseases. In models of emphysema or fibrosis, the essential ...

Generation of a Chronic Obstructive Pulmonary Disease Model in Mice by Repeated Ozone Exposure

Chronic obstructive pulmonary disease (COPD) is characterized by persistent airflow limitation and lung parenchymal destruction. It has a very high incidence in aging populations. The current conventional therapies for COPD focus mainly on symptom-modifying drugs; thus, the development of new therapies is urgently needed. Qualified animal models of COPD could help to characterize the underlying mechanisms and can be used for new drug screening. Current COPD models, such as lipopolysaccharide (LPS) or the porcine pancreatic elastase (PPE)-induced emphysema model, generate COPD-like lesions in the lungs and airways but do not otherwise resemble the pathogenesis of human COPD. A cigarette smoke (CS)-induced model remains one of the most popular because it not only simulates COPD-like lesions in the respiratory system, but it is also based on one of the main hazardous materials that causes COPD in humans. However, the time-consuming and labor-intensive aspects of the CS-induced model dramatically limit its application in new drug screening. In this study, we successfully generated a new COPD model by exposing mice to high levels of ozone. This model demonstrated the following: 1) decreased forced expiratory volume 25, 50, and 75/forced vital capacity (FEV25/FVC, FEV50/FVC, and FEV75/FVC), indicating the deterioration of lung function; 2) enlarged lung alveoli, with lung parenchymal destruction; 3) reduced fatigue time and distance; and 4) increased inflammation. Taken together, these data demonstrate that the ozone exposure (OE) model is a reliable animal model that is similar to humans because ozone overexposure is one of the etiological factors of COPD. Additionally, it only took 6 - 8 weeks, based on our previous work, to create an OE model, whereas it requires 3 - 12 months to induce the cigarette smoke model, indicating that the OE model might be a good choice for COPD research.

This study describes the successful generation of a new chronic obstructive pulmonary disease (COPD) animal model by repeatedly exposing mice to high concentrations of ozone.

반복된 오존 노출에 의해 쥐에 만성 폐쇄성 폐 질환 모델의 생성

Chronic obstructive pulmonary disease (COPD) is characterized by persistent airflow limitation and lung parenchymal destruction. It has a very high ...

The Left Pneumonectomy Combined with Monocrotaline or Sugen as a Model of Pulmonary Hypertension in Rats

In this protocol, we detail the correct procedural steps and necessary precautions to successfully perform a left pneumonectomy and induce PAH in rats with the additional administration of monocrotaline (MCT) or SU5416 (Sugen). We also compare these two models to other PAH models commonly used in research. In the last few years, the focus of animal PAH models has moved towards studying the mechanism of angioproliferation of plexiform lesions, in which the role of increased pulmonary blood flow is considered as an important trigger in the development of severe pulmonary vascular remodeling. One of the most promising rodent models of increased pulmonary flow is the unilateral left pneumonectomy combined with a "second hit" of MCT or Sugen. The removal of the left lung leads to increased and turbulent pulmonary blood flow and vascular remodeling. Currently, there is no detailed procedure of the pneumonectomy surgery in rats. This article details a step-by-step protocol of the pneumonectomy surgical procedure and post-operative care in male Sprague-Dawley rats. Briefly, the animal is anesthetized and the chest is opened. Once the left pulmonary artery, pulmonary vein, and bronchus are visualized, they are ligated and the left lung is removed. The chest then closed and the animal recovered. Blood is forced to circulate only on the right lung. This increased vascular pressure leads to a progressive remodeling and occlusion of small pulmonary arteries. The second hit of MCT or Sugen is used one week post-surgery to induce endothelial dysfunction. The combination of increased blood flow in the lung and endothelial dysfunction produces severe PAH. The primary limitation of this procedure is that it requires general surgical skills.

The rodent left pneumonectomy is a valuable technique in pulmonary hypertension research. Here, we present a protocol to describe the rat pneumonectomy procedure and postoperative care to ensure minimal morbidity and mortality.

쥐에 폐 고혈압의 모델로 Monocrotaline 또는 Sugen와 함께 왼쪽된 Pneumonectomy

In this protocol, we detail the correct procedural steps and necessary precautions to successfully perform a left pneumonectomy and induce PAH in rats ...

Experimental Model to Evaluate Resolution of Pneumonia

Acute respiratory distress syndrome (ARDS) causes acute lung injury, characterized by rapid alveolar damage and severe hypoxemia. This, in turn, leads to high morbidity and mortality. Currently, there are no pre-clinical models that recapitulate the complexity of human ARDS. However, infectious models of pneumonia (PNA) can replicate the main pathophysiological features of ARDS. Here, we describe a model of PNA induced by the intratracheal instillation of live Streptococcus pneumoniae and Klebsiella pneumoniae in C57BL6 mice. In order to evaluate and characterize the model, after inducing injury, we carried out serial measurements of body weight and bronchoalveolar lavage (BAL) for measuring markers of lung injury. Additionally, we harvested lungs for cell count and differentials, BAL protein quantification, cytospin, bacterial colony-forming unit counts, and histology. Lastly, high dimensional flow cytometry was performed. We propose this model as a tool to understand the immune landscape during the early and late resolution phases of lung injury.

This manuscript describes the establishment of an infectious model of pneumonia in mice and the respective characterization of injury resolution along with methods for growing bacteria and intratracheal instillation. A novel approach using high-dimensional flow cytometry to evaluate the immune landscape is also described.

폐렴의 해결을 평가하기 위한 실험모델

Acute respiratory distress syndrome (ARDS) causes acute lung injury, characterized by rapid alveolar damage and severe hypoxemia. This, in turn, leads to ...

면역학 및 감염병학

A Rat Lung Transplantation Model of Warm Ischemia/Reperfusion Injury: Optimizations to Improve Outcomes

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods for choosing appropriate cuff sizes for the pulmonary vein (PV), pulmonary artery (PA), or bronchus (Br) are varied, thus making the determination of proper cuff size during rat lung transplantation an exercise of trial and error. By standardizing the cuffing technique to use the smallest effective cuff appropriate for the size of the vessel or bronchus, one can make the transplantation procedure safer, faster, and more successful. Since diameters of the PV, PA, and Br are related to the body weight of the rat, we present a strategy to choosing an appropriate size using a weight-based guide. Since lung volume is also related to body weight, we recommend that this relationship should also be considered when choosing the proper volume of air for donor lung inflation during warm ischemia as well as for the proper volume of PBS to be instilled during bronchoalveolar lavage (BAL) fluid collection. We also describe methods for 4th intercostal space dissection, wound closure, and sample collection from both the native and transplanted lobes.

Here, we present optimizations to a rat lung transplantation model that serve to improve outcomes. We provide a size guide for cuffs based on body weight, a measurement strategy to ascertain the 4th intercostal space, and methods of wound closure and BAL (bronchoalveolar lavage) fluid and tissue collection.

온난 허혈/재관류 손상의 쥐 폐 이식 모델: 결과 개선을 위한 최적화

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods ...

Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: Evaluation of Lung Function and Metabolism

Lung transplantation (LTx) remains the standard of care for end-stage lung disease. A shortage of suitable donor organs and concerns over donor organ quality exacerbated by excessive geographic transportation distance and stringent donor organ acceptance criteria pose limitations to current LTx efforts. Ex situ lung perfusion (ESLP) is an innovative technology that has shown promise in attenuating these limitations. The physiologic ventilation and perfusion of the lungs outside of the inflammatory milieu of the donor body affords ESLP several advantages over traditional cold static preservation (CSP). There is evidence that negative pressure ventilation (NPV) ESLP is superior to positive pressure ventilation (PPV) ESLP, with PPV inducing more significant ventilator-induced lung injury, pro-inflammatory cytokine production, pulmonary edema, and bullae formation. The NPV advantage is perhaps due to the homogenous distribution of intrathoracic pressure across the entire lung surface. The clinical safety and feasibility of a custom NPV-ESLP device have been demonstrated in a recent clinical trial involving extender criteria donor (ECD) human lungs. Herein, the use of this custom device is described in a juvenile porcine model of normothermic NPV-ESLP over a 12 h duration, paying particular attention to management techniques. Pre-surgical preparation, including ESLP software initialization, priming, and de-airing of the ESLP circuit, and the addition of anti-thrombotic, anti-microbial, and anti-inflammatory agents, is specified. The intraoperative techniques of central line insertion, lung biopsy, exsanguination, blood collection, cardiectomy, and pneumonectomy are described. Furthermore, particular focus is paid to anesthetic considerations, with anesthesia induction, maintenance, and dynamic modifications outlined. The protocol also specifies the custom device's initialization, maintenance, and termination of perfusion and ventilation. Dynamic organ management techniques, including alterations in ventilation and metabolic parameters to optimize organ function, are thoroughly described. Finally, the physiological and metabolic assessment of lung function is characterized and depicted in the representative results.

Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: Evaluation of Lung Function and Metabolism

This paper describes a porcine model of negative pressure ventilation ex situ lung perfusion, including procurement, attachment, and management on the custom-made platform. Focus is made on anesthetic and surgical techniques, as well as troubleshooting.

Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: 폐 기능 및 대사 평가

Lung transplantation (LTx) remains the standard of care for end-stage lung disease. A shortage of suitable donor organs and concerns over donor organ ...

A Simple Cuff Technique for Murine Left Lung Transplantation

Over the past decade, our laboratory has made significant progress in developing and refining vascularized mouse lung transplantation models using an efficient and highly reliable "cuff technique" of transplantation. This article describes a sophisticated and comprehensive method for orthotopic lung transplantation in a vascularized orthotopic lung model, representing the most physiologic and clinically relevant model of mouse lung transplantation to date. The transplantation process consists of two distinct stages: donor harvest and subsequent implantation into the recipient. The method has been successfully mastered, and with several months of sufficient training, a skilled practitioner can perform the procedure in approximately 90 min from skin-to-skin. Surprisingly, once individuals overcome the initial learning curve, the survival rate during the perioperative period approaches nearly 100%. The mouse model allows for the use of multiple commercially available transgenic and mutant strains of mice, enabling the study of tolerance and rejection. Additionally, the unique features of this model make it a valuable tool for investigating tumor biology and immunology.

The present protocol describes a cuff technique for a mouse left lung transplantation model. This technique has been developed over several years and has performed well, serving effectively in immunological research.

Over the past decade, our laboratory has made significant progress in developing and refining vascularized mouse lung transplantation models using an ...

Ex Vivo Porcine Experimental Model for Studying and Teaching Lung Mechanics

Mechanical ventilation is widely used and requires specific knowledge for understanding and management. Health professionals in this field may feel insecure and lack knowledge because of inadequate training and teaching methods. Therefore, the objective of this article is to outline the steps involved in generating an ex vivo porcine lung model to be used in the future, to study and teach lung mechanics. To generate the model, five porcine lungs were carefully removed from the thorax following the guidelines of the Animal Research Ethics Committee with adequate care and were connected to the mechanical ventilator through a tracheal cannula. These lungs were then subjected to the alveolar recruitment maneuver. Respiratory mechanics parameters were recorded, and video cameras were used to obtain videos of the lungs during this process. This process was repeated for five consecutive days. When not used, the lungs were kept refrigerated. The model showed different lung mechanics after the alveolar recruitment maneuver every day; not being influenced by the days, only by the maneuver. Therefore, we conclude that the ex vivo lung model can provide a better understanding of lung mechanics and its effects, and even of the alveolar recruitment maneuver through visual feedback during all stages of the process.