Name: angiogenesis in the ischemic rat lung
Uploaded: 2013-02-08T13:00:00
Description: Watch this Scientific Journal Video about Angiogenesis in the Ischemic Rat Lung at JoVE.com

우리는 폐의 캐스팅에 지원 한 박사 Adlah Sukkar, MD의 작품을 인정합니다. 이 작품은 NHLBI, HL088005로 운영되었습니다.

쥐에서 수행 모든 프로토콜은 존스 홉킨스 대학 동물 관리 및 사용위원회와 NIH의 지침에 따라 승인되었습니다. 가능하면 동물은 수술 외과 사이트의 오염을 최소화하기 위해 수술 영역에서 별도의 지역에 준비해야합니다. 1. 마취 / 무통 <ol><li> 3퍼센트 isoflurane과 함께 주입 유도 챔버에서, 장소 쥐 (할렌, 인디애나 폴리스, IN Sprague Dawley 남성의 쥐, 125~150그램). </li><li> 3퍼센?...

<ol>
	<li>Weibel, E. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+stages+in+the+development+of+collateral+circulation+to+the+lung+in+the+rat.">Early stages in the development of collateral circulation to the lung in the rat.</a> Circulation Research. 8, 353-376 (1960).</li><li>Li, X., Wilson, 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=Increased+vascularity+of+the+bronchial+mucosa+in+mild+asthma.">Increased vascularity of the bronchial mucosa in mild asthma.</a> Am. J. Respir. Crit. Care Med. 156, 229-233 (1997).</li><li>Turner-Warwick, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Precapillary+systemic-pulmonary+anastomoses.">Precapillary systemic-pulmonary anastomoses.</a> Thorax. 18, 225-237 (1963).</li><li>Muller, K. M., Meyer-Schwickerath, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bronchial+arteries+in+various+stages+of+bronchogenic+carcinoma.">Bronchial arteries in various stages of bronchogenic carcinoma.</a> Pathol. Res. Pract. 163, 34-46 (1978).</li><li>Remy-Jardin, M., Duhamel, 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=Systemic+Collateral+Supply+in+Patients+with+Chronic+Thromboembolic+and+Primary+Pulmonary+Hypertension:+Assessment+with+Multi-Detector+Row+Helical+CT+Angiography.">Systemic Collateral Supply in Patients with Chronic Thromboembolic and Primary Pulmonary Hypertension: Assessment with Multi-Detector Row Helical CT Angiography.</a> Radiology. , 274-281 (2005).</li><li>Karsner, H., Ghoreyeb, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studies+in+infarction:+The+circulation+in+experimental+pulmonary+embolism.">Studies in infarction: The circulation in experimental pulmonary embolism.</a> J. Exp. Med. 18, 507-522 (1913).</li><li>Endrys, J., Hayat, N., 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=Comparison+of+bronchopulmonary+collaterals+and+collateral+blood+flow+in+patients+with+chronic+thromboembolic+and+primary+pulmonary+hypertension.">Comparison of bronchopulmonary collaterals and collateral blood flow in patients with chronic thromboembolic and primary pulmonary hypertension.</a> Heart. 78, 171-176 (1997).</li><li>Virchow, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uber+die+Standpunkte+in+den+Wissenschaftlichen+Medizin.">Uber die Standpunkte in den Wissenschaftlichen Medizin.</a> Virchow Archiv. 1, 1-19 .</li><li>Fadel, E., Mazmanian, G. 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=Lung+reperfusion+injury+after+chronic+or+acute+unilateral+pulmonary+artery+occlusion.">Lung reperfusion injury after chronic or acute unilateral pulmonary artery occlusion.</a> Am. J. Respir. Crit. Care Med. 157, 1294-1230 (1998).</li><li>Charan, N. B., Carvalho, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Angiogenesis+in+bronchial+circulatory+system+after+unilateral+pulmonary+artery+obstruction.">Angiogenesis in bronchial circulatory system after unilateral pulmonary artery obstruction.</a> J. Appl. Physiol. 82, 284-291 (1997).</li><li>Shi, W., Hu, F., 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=Altered+reactivity+of+pulmonary+vessels+in+postobstructive+pulmonary+vasculopathy.">Altered reactivity of pulmonary vessels in postobstructive pulmonary vasculopathy.</a> J. Appl. Physiol. 88, 17-25 (2000).</li><li>Shi, W., Giaid, 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+reactivity+to+endothelin+of+pulmonary+arteries+in+long-term+post-obstructive+pulmonary+vasculopathy+in+rats.">Increased reactivity to endothelin of pulmonary arteries in long-term post-obstructive pulmonary vasculopathy in rats.</a> Pulm. Pharmacol. Ther. 11, 189-196 (1998).</li><li>Sukkar, A., Jenkins, J., 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=Inhibition+of+CXCR2+Attenuates+Bronchial+Angiogenesis+in+the+Ischemic+Rat+Lung.">Inhibition of CXCR2 Attenuates Bronchial Angiogenesis in the Ischemic Rat Lung.</a> J. Appl. Physiol. 104, 1470-1475 (2008).</li><li>Mitzner, W., Lee, 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=Angiogenesis+in+the+mouse+lung.">Angiogenesis in the mouse lung.</a> Am. J. Pathol. 157, 93-101 (2000).</li><li>Wagner, E. M., Jenkins, J., 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=Lung+and+vascular+function+during+chronic+severe+pulmonary+ischemia.">Lung and vascular function during chronic severe pulmonary ischemia.</a> J. Appl. Physiol. 110, 538-544 (2011).</li><li>Remy-Jardin, M., Bouaziz, N., 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=Bronchial+and+nonbronchial+systemic+arteries+at+multi-detector+row+CT+angiography:+comparison+with+conventional+angiography.">Bronchial and nonbronchial systemic arteries at multi-detector row CT angiography: comparison with conventional angiography.</a> Radiology. 233, 741-749 (2004).</li><li>Baluk, P., Tammela, 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=Pathogenesis+of+persistent+lymphatic+vessel+hyperplasia+in+chronic+airway+inflammation.">Pathogenesis of persistent lymphatic vessel hyperplasia in chronic airway inflammation.</a> J. Clin. Invest. 115, 247-257 (2005).</li><li>Bailey, S. R., Boustany, S., 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=Airway+vascular+reactivity+and+vascularisation+in+human+chronic+airway+disease.">Airway vascular reactivity and vascularisation in human chronic airway disease.</a> Pulm. Pharmacol. Ther. 22, 417-425 (2009).</li></ol>

테스트 모든 종의 좌측 폐동맥의 결합은 허혈성 폐의 강력한 조직 neovascularization로 연결됩니다. 우리는 쥐 모델에서 수술 방법의 세부 사항을 제시하고 있습니다. 우리의 결과는 혈관 주조, 조직 병리학에서 생산 및 생체 라벨에 기관지 동맥 세포 분열 따위에 의해 번식을 명시하고 폐 실질을 perfuse. 따라서, 기관지 혈관 신생의 메커니즘은 만성 폐 thromboembolism의 인간의 상태를 패러랠 동물 ...

폐의 조직 혈관 신생은 잘 인식됩니다. 이러한 천식 2, 간질 폐 섬유증이 3 암 4 및 만성 폐 thromboembolism 5의 전신 vasculature과 폐 실질 폐 proliferates 주변과 침공과 같은 질병 상태에서. 그러나, 조직의 차동 활성화보다는 폐 순환을 공부하는 동물 모델은 몇 가지 있습니다. 아마도 성인 포유 동물의 폐에 조직 neovascularization의 가장 재현 모델은 만성 폐 동맥 국소 빈혈을 유도 한 후 발생하는 것입니다. 5-7 인간 좌측 폐동맥 폐쇄에 대한 반응은, 개 8, 돼지 9, 양 10, 기니 돼지 11, 쥐 1, 12, 13 및 마우스 14 기관지 동맥뿐만 아니라 갈비 사이 동맥의 급속한 확산이다. 전신 neovasculari에 대한 책임 메커니즘폐 국소 빈혈 후 폐의 zation은 대부분 알 수없는 널리 연구되지 ​​않았습니다. 좌측 폐동맥 폐쇄 후 쥐의 기관지 혈관 신생의 시간 코스는 신중하게 Weibel 1의 histologic 작품에 설명되어 있습니다. 쥐에이 작품 확장, 저희 연구실이 과정에서 중요한 성장 요인뿐만 아니라 폐에서이 neovasculature의 physiologic 결과 모두에 초점을 맞추고있다. 결과가 CXC 케모카인의 CINC-3은 초기 국소 빈혈 후에 CXCR2에 중화 항체를 쥐를 치료 올라간다 보여, CINC-3 수용체는 혈관 신생 13 attenuates. 십사일 폐 국소 빈혈의 발병 후 새로 설립 된 기관지 vasculature 수가 크게 증가 단백질 투자율 15 이상으로 표시했습니다. 왼쪽 폐 기능은 diffusing 용량과 폐 볼륨 15 감소 감소 일반 게재되지 않았습니다. neovasculature가 공동있을 수 있습니다 있지만만성 폐 국소 빈혈 동안 폐 조직의 보존에 ntributed, 그것은 정상하고 폐 기능의 지속적인 감소에 기여할 수는 없습니다 나타납니다. 아마도이 모델의 가장 호기심 측면 중 하나는 혈관을 proliferating의 공간적 분포에 관한 것이다. 국소 빈혈로 인한 폐 실질 조직 내에서의 성장 요인의 출시에도 불구하고, neovasculature이 상대적으로 큰 업스트림 기관지 동맥에서 유래 된 것입니다. 정상적인 기관지 동맥은 대동맥에서 작은 지점으로 발생하고 카리나의기도 트리를 침공. 따라서 성장 요인 arteriogenesis의 초기 단계를 유도하는하여 메커니즘은 명확하지 않습니다. 우리는 쥐가 인간과 비슷한 혈관 해부학 적으로 폐 국소 빈혈 동안 전신 혈관 신생을 담당하는 메커니즘을 연구 할 수있는 독특한 기회를 제공하는 것이 좋습니다. 좌측 폐동맥의 완전한 방해가 인간의 과목에서 드문이지만하면,증가 기관지 vascularity는 유사 환자에서 유도 할 것으로 보인다 폐동맥 폐쇄 (16)의 어떤 사이트와 크기입니다. 따라서, 우리는 쥐의 왼쪽 폐 동맥과 혈관 신생의 정도를 수량화 할 수있는 수단을 ligate 할 수있는 수술 방법에 대한 자세한 설명을 제공합니다.

<table border="1"><tbody><tr><td> 시약 : </td><td> 회사 </td><td></td><td></td></tr><tr><td> buprenorphine 히드로 클로라이드, Puralube </td><td> 버틀러 Schein </td><td></td><td></td></tr><tr><td> bupivicaine </td><td> APP 제약 </td><td></td><td></td></tr><tr><td> Povidone - 요오드 swabstick </td><td> Dynarex 공사 </td><td></td><td></td></tr><tr><td> 폴리 프로필렌의 봉합 크기 6 대 0으로 8분의 3 원 역 절단 바늘 </td><td> Myco 의료 </td><td></td><td></td></tr><tr><td> PE20 튜브 </td><td> Becton 디킨슨 </td><td></td><td></td></tr><tr><td> 15 μm 진홍의 폴리스티렌 fluorospheres </td><td> Invitrogen </td><td></td><td></td></tr><tr><td> 한 ML 해밀턴 유리 주사기 </td><td> 해밀턴 회사 </td><td> &lt;/ TD&gt; <td></td></tr><tr><td> 장비 : </td><td></td><td></td><td></td></tr><tr><td> 램프의 요정 플러스 주사기 펌프 </td><td> 켄트 과학 </td><td></td><td></td></tr><tr><td> 형광 분광 </td><td> Digilab </td><td></td><td></td></tr><tr><td> 쥐 환풍기 모델 683 </td><td> 하버드 장치 </td><td></td><td></td></tr><tr><td></td><td></td><td></td><td> 표 1. 특정 시약 및 장비 표. </td></tr></tbody></table>

angiogenesis in the ischemic rat lung

성인 폐는 전신 기관지 동맥과 폐동맥 동맥을 통해 흐르는 전체 정맥 반환 모두에 의해 perfused 있습니다. 대부분의 폐 pathologies에서는 향상된 폐 관류에 대한 필요에 응답하고 강력한 neovascularization을 보여주는 작은 조직 vasculature입니다. 폐 동맥 방해에 의해 유도 폐 혈관 국소 빈혈은 사람의 급속한 전신 동맥 혈관 신생뿐만 아니라 여러 동물 모델에서 결과로 표시되었습니다. 쥐의 기관지 동맥 확산의 시간 코스의 histologic 평가는 신중하게 Weibel 1 설명되었지만, 새로운 혈관의 조직 성장을위한 책임이 메커니즘은 명확하지 않습니다. 우리는 기관지 neovascularization로 연결 쥐에서 좌측 폐동맥의 국소 빈혈을 일으킬 외과 세부 정보를 제공합니다. 혈관 신생의 정도의 정량화는 폐 내의 두 혈관 침대의 존재로 인해 추가 과제를 제공합니다. 방법표시 microsphere 주사에 따라 기능 혈관 신생을 결정하기 위해 제공됩니다.

혈관 캐스트 : 쥐의 좌측 폐동맥의 국소 빈혈의 효과의 결과는 그림 1에 설명되어 있습니다. 표시하면 기관지 vasculature의 메타 크릴 레이트 캐스트와 28일 LPAL 후 왼쪽기도 트리의 광범위한 vascularity입니다. 이 캐스트를 얻으려면 조직 vasculature는 내림차순으로 대동맥에 역행, 메타 크릴 레이트 혼합물 (빨간색)로 주입하였으며, 기관은 cannulated과 흰색 실리콘 기반 소재로 주입되었다. ?...

Watch this Scientific Journal Video about Angiogenesis in the Ischemic Rat Lung at JoVE.com

Angiogenesis in the Ischemic Rat Lung

폐는 전신 기관지 동맥과 폐동맥 동맥 모두에서 perfused 있습니다. 대부분의 폐 pathologies에서는 강력한 neovascularization을 보여주는 작은 조직 vasculature입니다. 폐 혈액 흐름의 중단이 활발 기관지 혈관 신생을 촉진합니다. 우리는 기관지 neovascularization을 촉진 좌측 폐동맥의 국소 빈혈을 일으킬 외과 세부 정보를 제공합니다.

허혈성 쥐 폐의 혈관 신생

The  adult lung is perfused by both the systemic bronchial artery and the entire  venous return flowing through the pulmonary arteries. In most lung ...

Research

JoVE Journal

Medicine

Multi-modal Imaging of Angiogenesis in a Nude Rat Model of Breast Cancer Bone Metastasis Using Magnetic Resonance Imaging, Volumetric Computed Tomography and Ultrasound

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell proliferation in the bone marrow cavity as well as for interaction of tumor and bone cells resulting in local bone destruction. Our aim was to develop a model of experimental bone metastasis that allows in vivo assessment of angiogenesis in skeletal lesions using non-invasive imaging techniques.
For this purpose, we injected 105 MDA-MB-231 human breast cancer cells into the superficial epigastric artery, which precludes the growth of metastases in body areas other than the respective hind leg1. Following 25-30 days after tumor cell inoculation, site-specific bone metastases develop, restricted to the distal femur, proximal tibia and proximal fibula1. Morphological and functional aspects of angiogenesis can be investigated longitudinally in bone metastases using magnetic resonance imaging (MRI), volumetric computed tomography (VCT) and ultrasound (US).
MRI displays morphologic information on the soft tissue part of bone metastases that is initially confined to the bone marrow cavity and subsequently exceeds cortical bone while progressing. Using dynamic contrast-enhanced MRI (DCE-MRI) functional data including regional blood volume, perfusion and vessel permeability can be obtained and quantified2-4. Bone destruction is captured in high resolution using morphological VCT imaging. Complementary to MRI findings, osteolytic lesions can be located adjacent to sites of intramedullary tumor growth. After contrast agent application, VCT angiography reveals the macrovessel architecture in bone metastases in high resolution, and DCE-VCT enables insight in the microcirculation of these lesions5,6. US is applicable to assess morphological and functional features from skeletal lesions due to local osteolysis of cortical bone. Using B-mode and Doppler techniques, structure and perfusion of the soft tissue metastases can be evaluated, respectively. DCE-US allows for real-time imaging of vascularization in bone metastases after injection of microbubbles7.
In conclusion, in a model of site-specific breast cancer bone metastases multi-modal imaging techniques including MRI, VCT and US offer complementary information on morphology and functional parameters of angiogenesis in these skeletal lesions.

In the pathogenesis of bone metastasis, angiogenesis is a crucial process and therefore represents a target for imaging and therapy. Here, we present a rat model of site-specific breast cancer bone metastasis and describe strategies to non-invasively image angiogenesis in vivo using magnetic resonance imaging, volumetric computed tomography and ultrasound.

자기 공명 영상, 체적 계산의 Tomography 및 초음파를 이용한 유방암의 뼈 전이의 누드 쥐 모델에서 Angiogenesis의 멀티 모달 이미징

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell ...

연구

의학

Retinal Detachment Model in Rodents by Subretinal Injection of Sodium Hyaluronate

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the occurrence of subretinal hemorrhage can affect photoreceptor cell death in the detached retina. Hence, it is advantageous to create reproducible RDs without subretinal hemorrhage for evaluating photoreceptor cell death. We modified a previously reported method to create bullous and persistent RDs in a reproducible location with rare occurrence of subretinal hemorrhage. The critical step of this modified method is the creation of a self-sealing scleral incision, which can prevent leakage of sodium hyaluronate after injection into the subretinal space. To make the self-sealing scleral incision, a scleral tunnel is created, followed by scleral penetration into the choroid with a 30 G needle. Although choroidal hemorrhage may occur during this step, astriction with a surgical spear reduces the rate of choroidal hemorrhage. This method allows a more reproducible and reliable model of photoreceptor death in diseases that involve RD such as rhegmatogenous RD, retinopathy of prematurity, diabetic retinopathy, central serous chorioretinopathy, and age-related macular degeneration (AMD).

Creating experimental retinal detachments with a reproducible and sustained height of detachment, and without subretinal hemorrhage, is important for studying the pathophysiology of photoreceptor cell loss in retinal disease and evaluating potential therapeutic interventions. Here, we report such a method in detail.

나트륨 히알루 론산의 망막 주사 설치류에있는 망막 박리 모델

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the ...

Implantation of Fibrin Gel on Mouse Lung to Study Lung-specific Angiogenesis

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and repair damage in simple tissues in the orthopedic and periodontal fields. However, attempts to regenerate the structures and functions of more complex three-dimensional (3D) organs such as lungs have not been very successful because the biological processes of organ regeneration have not been well explored. It is becoming clear that angiogenesis, the formation of new blood vessels, plays key roles in organ regeneration. Newly formed vasculatures not only deliver oxygen, nutrients and various cell components that are required for organ regeneration but also provide instructive signals to the regenerating local tissues. Therefore, to successfully regenerate lungs in an adult, it is necessary to recapitulate the lung-specific microenvironments in which angiogenesis drives regeneration of local lung tissues. Although conventional in vivo angiogenesis assays, such as subcutaneous implantation of extracellular matrix (ECM)-rich hydrogels (e.g., fibrin or collagen gels or Matrigel - ECM protein mixture secreted by Engelbreth-Holm-Swarm mouse sarcoma cells), are extensively utilized to explore the general mechanisms of angiogenesis, lung-specific angiogenesis has not been well characterized because methods for orthotopic implantation of biomaterials in the lung have not been well established. The goal of this protocol is to introduce a unique method to implant fibrin gel on the lung surface of living adult mouse, allowing for the successful recapitulation of host lung-derived angiogenesis inside the gel. This approach enables researchers to explore the mechanisms by which the lung-specific microenvironment controls angiogenesis and alveolar regeneration in both normal and pathological conditions. Since implanted biomaterials release and supply physical and chemical signals to adjacent lung tissues, implantation of these biomaterials on diseased lung can potentially normalize the adjacent diseased tissues, enabling researchers to develop new therapeutic approaches for various types of lung diseases.

Recapitulation of the organ-specific microenvironment, which stimulates local angiogenesis, is indispensable for successful regeneration of damaged tissues. This report demonstrates a novel method to implant fibrin gels on the lung surface of living mouse in order to explore how the lung-specific microenvironment modulates angiogenesis and alveolar regeneration in adult mouse.

마우스 폐에 섬유소 젤의 주입은 폐 관련 신생 혈관 연구에

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and ...

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. As this technology expands and improves, the ability to potentially evaluate and improve the quality of substandard lungs prior to transplant is a critical need. In order to more rigorously evaluate these approaches, a reproducible animal model needs to be established that would allow for testing of improved techniques and management of the donated lungs as well as to the lung-transplant recipient. In addition, an EVLP animal model of associated pathologies, e.g., ventilation induced lung injury (VILI), would provide a novel method to evaluate treatments for these pathologies. Here, we describe the development of a rat EVLP lung program and refinements to this method that allow for a reproducible model for future expansion. We also describe the application of this EVLP system to model VILI in rat lungs. The goal is to provide the research community with key information and &#8220;pearls of wisdom&#8221;/techniques that arose from trial and error and are critical to establishing an EVLP system that is robust and reproducible.

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. Here, we describe the development of a rat EVLP program and refinements that allow for a reproducible model for future expansion.

고립의 방법<em&gt; 전의 VIVO</em쥐 모델에서&gt; 폐 관류 : 쥐 EVLP 프로그램 개발에서 교훈을

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed ...

Rat Model of Photochemically-Induced Posterior Ischemic Optic Neuropathy

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history have&#160;remained poorly understood. Recently, we developed a reliable, reproducible animal model of PION and tested the treatment effect of some neurotrophic factors in this model1. The purpose of this video is to demonstrate our photochemically induced model of posterior ischemic optic neuropathy, and to evaluate its effects with retrograde labeling of retinal ganglion cells. Following surgical exposure of the posterior optic nerve, a photosensitizing dye, erythrosin B, is intravenously injected and a laser beam is focused onto the optic nerve surface. Photochemical interaction of erythrosin B and the laser during irradiation damages the vascular endothelium, prompting microvascular occlusion mediated by platelet thrombosis and edematous compression. The resulting ischemic injury yields a gradual but pronounced retinal ganglion cell dieback, owing to a loss of axonal input - a remote, injury-induced and clinically relevant outcome. Thus, this model provides a novel platform to study the pathophysiologic course of PION,&#160;and can be further optimized for testing therapeutic approaches for optic neuropathies as well as other CNS ischemic diseases.

The goal of this protocol is to photochemically induce ischemic injury to the posterior optic nerve in rat. This model is critical to studies of the pathophysiology of posterior ischemic optic neuropathy, and therapeutic approaches for this and other optic neuropathies, as well as of other CNS ischemic diseases.

광 화학적으로 유도 된 후방 허혈성 시신경 병증의 쥐 모델

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history ...

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 ...

Demonstration of the Rat Ischemic Skin Wound Model

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and unrelieved pressure due to immobility. Animal models have been developed that attempt to mimic these conditions for the purpose of furthering our understanding of the complexity of chronic wounds. The model described herein is a rat ischemic skin flap model that permits a prolonged reduction of blood flow resulting in wounds that become ischemic and resemble a chronic wound phenotype (reduced vascularization, increased inflammation and delayed wound closure). It consists of a bipedicled dorsal flap with 2 ischemic wounds placed centrally and 2 non-ischemic wounds lateral to the flap as controls. A novel addition to this ischemic skin flap model is the placement of a silicone sheet beneath the flap that functions as a barrier and a splint to prevent revascularization and reduce contraction as the wounds heal. Despite the debate of using rats for wound healing studies due to their quite distinct anatomic and physiologic differences compared to humans (i.e., the presence of a panniculus carnosus muscle, short life-span, increased number of hair follicles, and their ability to heal infected wounds) the modifications employed in this model make it a valuable alternative to previously developed ischemic skin flap models.

The rat, due to its size, availability, and rather docile behavior, has been utilized as a research model for many years. The goal of this protocol is to utilize the rat as an ischemic skin wound healing model to provide valuable insight into the pathophysiology of chronic wounds.

쥐 허혈성 피부 상처 모델의 데모

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and ...

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 ...

Securing the Rat in the Rat Jacket for Electroacupuncture

Wear a pair of gloves before proceeding to fit and secure the rat in the custom made rat jacket. Then using one hand hold the rat jacket's middle and back portion on the table. Using the other hand, gently hold the rat by its tail to pick it up from the feeding cage and place it into the jacket as the rat moves forward into the jacket and its head reaches the headgear point.Using both hands, carefully pull out its left and right upper limbs from the holes in the jacket. Securely wrap the tying ropes from the opening end of the jacket around the rat's trunk, and tie them tightly at the opposite end to adequately restrain its upper limbs. Tie the rope loose enough to prevent hypoxia or local limb ischemia, and tight enough to prevent the rat's escape.In a similar manner, secure its lower limbs.

전기 침술을 위해 쥐 재킷에 쥐를 고정합니다.

<meta charset="utf8"></head><body>생쥐 폐 이식에서 세포 역학의 실시간 모니터링

Intravital Two-Photon Microscopy of the Transplanted Mouse Lung

Complications after lung transplantation are largely related to the host immune system responding to the graft. Such immune responses are regulated by crosstalk between donor and recipient cells. A better understanding of these processes relies on the use of preclinical animal models and is aided by an ability to study intra-graft immune cell trafficking in real-time. Intravital two-photon microscopy can be used to image tissues and organs for depths up to several hundred microns with minimal photodamage, which affords a great advantage over single-photon confocal microscopy. Selective use of transgenic mice with promoter-specific fluorescent protein expression and/or adoptive transfer of fluorescent dye-labeled cells during intravital two-photon microscopy allows for the dynamic study of single cells within their physiologic environment. Our group has developed a technique to stabilize mouse lungs, which has enabled us to image cellular dynamics in na&#239;ve lungs and orthotopically transplanted pulmonary grafts. This technique allows for detailed assessment of cellular behavior within the vasculature and in the interstitium, as well as for examination of interactions between various cell populations. This procedure can be readily learned and adapted to study immune mechanisms that regulate inflammatory and tolerogenic responses after lung transplantation. It can also be expanded to the study of other pathogenic pulmonary conditions.

<meta charset="utf8"></head><body>저자 스포트라이트: 생체 내 이미징을 통한 폐 이식 면역학의 발전

Here, we present a protocol to intravitally image the transplanted mouse left lung using two-photon microscopy. This represents a valuable tool for studying cellular dynamics and interactions in real-time following murine lung transplantation.

이식된 마우스 폐의 생체 내 2광자 현미경

Complications after lung transplantation are largely related to the host immune system responding to the graft. Such immune responses are regulated by ...