我们认识到，工作堡，的博士Adlah的MD协助铸造的肺。这项工作已经由NHLBI，HL088005。

对老鼠进行的所有协议，由美国约翰·霍普金斯大学动物护理和使用委员会批准，并按照与美国国立卫生研究院的指导方针。只要有可能，动物应该是手术准备在从手术区域分开的区域的外科手术部位，以尽量减少污染。 1。麻醉/镇痛<ol><li>放置在感应腔注入3％异氟醚的大鼠（只SD雄性大鼠，125-150克;哈伦，印第安纳波利斯，IN）。 </li><li>将板的鼻锥和呼吸机连接到3％异氟醚麻醉手术麻醉大鼠。使用无菌技术的所有程序。悬垂老鼠，以确保无菌操作领域。 </li></ol> 2。插管法<ol><li>仰卧位，用医用胶带固定附加物。 </li><li>删除大鼠的鼻锥，延长舌与软垫钳。 </li><li>使用14号intracath与钝的金属stilet的为导向。滑动后面ngue，进气管。 </li><li>取出金属stilet离开的白色塑料intracath在气管。确保将大鼠呼吸和空气流过管子。 </li><li>更换鼻锥直接转接器导管，连接呼吸机（90次/分，潮气量8毫升/公斤;啮齿动物呼吸机型号683，哈佛装置，州Holliston，马萨诸塞州）的老鼠。 </li><li>应用Puralube（巴特勒施恩​​，都柏林，OH）兽医药膏眼睛。 </li></ol> 3。开胸手术<ol><li>地点大鼠右侧下来，用医用胶带固定附属物。 </li><li>剃须左侧胸腔区。 </li><li>删除多余的皮毛，小真空。 </li><li>聚维酮碘棉棒（Dynarex公司，Orangebur，NY），其次是用酒精擦拭消毒领域。这个过程重复2次（共3磨砂）。 </li><li>横切口用无菌无菌手术刀解剖剪刀或在中心场。 </li><li>钝性解剖，通过L艾尔斯的组织和脂肪肋骨（最后一层薄膜覆盖肋骨）。 </li><li>计数肋骨，以确定第三肋间。 </li><li> 45°格雷夫使用无菌镊子，3 次和第 4肋骨之间切开钝。 </li><li>将肋骨分离器，拉轻轻地创建开放作废，完全可视化的肺，在缝合的地方磁带保持开放。 </li></ol> 4。左肺动脉结扎术<ol><li> 90°格雷夫钳，将左肺用右手。 </li><li>使用杜蒙模式第5直线镊子，用左手抓住左肺动脉和呼吸道。左肺动脉，将为在气道。确保镊子直接垂直于工作台。此外，它是最有效的在最前端的位置（最近的实质）拿起左肺动脉/左主支气管。将左肺外的通风这左手机动的。 </li><li>使用杜蒙模式＃5 45°弯钳分离左肺动脉，在其自然边界的左主支气管。这种分离出现薄白线之间的两个独立的结构。 </li><li>分离直接下动脉，而无需通过容器;顺畅地滑动沿分离的钳子提示保持在一起。 </li><li>继续直到提示钳明显分开，左肺动脉和左主支气管。只有一小点的提示，需要通过充分。如果血液可以可视化的钳子的前端上，然后它是不能完全通过和不应该尝试结扎。一旦通过的空间，左肺动脉，将为在曲线上的钳子。保持在这个位置。 </li><li>轻轻松开直钝镊子手柄（左手）和抢一块的预切缝（2-3英寸聚丙烯缝线大小6-0 MYCO医疗，卡里，NC）。 </li><li>打开弯钳抱着左肺动脉和g拉布缝合。通过空间之间的左肺动脉和左主支气管向上的相对运动的曲线的钳子轻轻地拉缝合。 </li><li>系紧阻断左肺动脉与一个平结，仔细剪断剩余的缝合线。 </li><li>关闭肋骨4-0安装到19毫米，3/8圈反向切割止血针（MYCO医疗，卡里，NC），小心不要使用钝钳按住肋骨和缝合用聚丙烯（蓝色单丝）尺寸缝合皮肤（仅肋骨）。 </li><li>完成一个宽松的平结，肺膨胀，积极呼气末正压（PEEP 2-5 CMH 2 O），hyperinflate肺当时的安全结得紧紧的，另一个结前剪断缝合其余的。移除PEEP和可视化，持续30秒，以确保，肺不折叠。将5滴布比卡因（APP制药，Schaumbur，IL）。 </li><li>关闭皮肤组织胶对伤口和皮肤一起推动使用返回END棉棒涂抹。给布比卡因（2.0毫克/公斤皮下）在切口部位随后每8小时，24小时或直到动物恢复正常活动。作为在这方面的皮下组织层是非常薄的，它是不容易在其自己的缝合。当组织胶施加和皮肤被推到一起，它也会关闭皮下层。 </li><li>关闭异氟醚气体，但继续的老鼠通风室内空气1-2分钟，直到自愿运动的回报。脱离呼吸机，拔除气管导管，确保将大鼠自主呼吸，然后再将其取出。 </li><li>擦去Puralube眼睛用棉签和监控运动和恢复。注射盐酸丁丙诺啡（0.05 mg / kg腹腔，巴特勒施恩​​，都柏林OH）。继续48小时手术后镇痛，每12小时交付。 </li></ol> 5。左侧颈总动脉插管要评估的幅度支气管perfus离子在期望的时间点后，左肺动脉结扎缺血左肺，注射标记的微球通过左侧颈动脉到主动脉弓。准备大鼠以上1-2。 <ol><li>切中线，沿着颈部中，冲解剖发现气管和左侧颈总动脉（微球注射部位）。 </li><li>插入导管充满肝素盐水，钝尖PE20管（Becton Dickinson公司，火花MD）连接到25克针，4路，1毫升注射器活塞的容器。 </li><li>将一瓶的微球（15μm的深红色聚苯乙烯荧光微球，1×10 6个球/毫升; Invitrogen公司，尤金，OR）在水中的超声波发生器30秒。 </li><li>取出瓶，涡和玻璃注射器1毫升汉密尔顿（Hamilton公司，雷诺，内华达州）至20克针吸取0.5毫升（500,000微球）。 </li><li>将4路活塞的汉密尔顿注射器注入微球注射泵（率：500μl/ min的精灵此外，肯特科学，Torringt上，CT）。 </li><li>移除Hamilton注射器和冲洗装置用1毫升heparized生理盐水，在500μl/ min的。 </li><li>执行完整的胸部开胸切断对大鼠下腔静脉抽血。 </li><li>拆下左侧的肺和其他组织的利益。 </li><li>要提取从组织中的微球，采取整个左肺后大鼠放血2M KOH（4-6毫升）中，并将其放置。在55°C水浴和离开组织消化过夜。加入吐温80（0.25％）洗珠，涡旋（10秒），并离心（2,000 rpm转速，20℃，10分钟）。除去上清液，添加2 - 乙氧基乙基乙酸酯（1毫升），涡旋，并静置1小时。后离心，离心（2000转;（20°C，10分钟），取出2 - 乙氧基乙酯水层含有的荧光，放置在试管和措施，使用日立F-2500型荧光分光光度计（激发612/emission 618 Digilab，州Holliston，MA）。 </li></ol>

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没有利益冲突的声明。

左肺动脉结扎在所有物种进行测试导致的缺血性强大的全身新生血管的肺。我们已经提出了详细的手术方法在大鼠模型。我们的研究结果所产生血管铸型，组织病理学，并在体内标记表明支气管动脉增殖和灌注肺实质。因此，的支气管血管生成机制，可以研究平行的人类生存条件的慢性肺血栓栓塞症的动物模型。此外，临床上相关的小动物模型提供了一个机会，学习的形式和功能的新生血?...

全身血管生成的肺是大家公认的。在疾病状态下，如哮喘，肺间质纤维化3，癌症，慢性肺血栓栓塞症，全身血管和周围的肺中繁衍，侵入肺实质。然而，动物模型来研究这种差别激活的系统，而不是肺循环的毕竟是少数。也许全身新生血管在成年哺乳动物的肺是最可重复性的模型是发生诱导慢性肺动脉缺血后。左肺动脉阻塞在人类5-7，狗，猪，羊，豚鼠，大鼠，12日，13日和小鼠14支气管动脉和肋间动脉的快速普及。全身neovasculari负责的机制zation的肺肺缺血后，在很大程度上是未知的，并没有被广泛研究。支气管左肺动脉阻塞后的大鼠血管生成的过程中的时间已被仔细描述在病理工作威贝尔1。这项工作在大鼠延伸的，我们的实验室已集中生长的重要因素，以及在这个过程中的生理结果本新生血管在肺。结果表明缺血后早期和中和抗体治疗大鼠CXCR2的趋化因子CINC-3升高，CINC-3受体，抑制血管新生13。新成立的支气管肺缺血发作后的14天内被证明是血管异常显着提高蛋白质的通透性15。左肺功能弥散能力下降和减少肺容积15中的表现不正常。虽然新生血管可能有合作ntributed慢性肺缺血肺组织的保护，它似乎不正常，可能会导致肺功能持续下降。 这种模式最奇怪的方面之一可能涉及血管增殖的空间分布。尽管在肺实质因缺血而释放生长因子，新生血管起源于比较大的上游支气管动脉。出现正常的支气管动脉从主动脉的一个小分支和侵入呼吸道树，在气管隆突。这样的机制，其中生长因子诱导的初始阶段动脉生成是不明确的。我们建议，大鼠，用类似人类的血管解剖，提供一个独特的机会来研究负责全身血管生成在肺缺血的机制。虽然左肺动脉完全阻塞是一种罕见的发生在人类的主题，增加支气管血管患者出现同样引起任何部位和大小的肺动脉阻塞16。因此，我们提供了手术结扎左肺动脉在大鼠和的一种手段，以量化的大小的血管生成的方法的详细描述。

<table border="1"><tbody><tr><td> 试剂： </td><td> 公司 </td><td></td><td></td></tr><tr><td>盐酸丁丙诺啡，Puralube; </td><td>巴特勒施恩</td><td></td><td></td></tr><tr><td>布比卡因</td><td> APP制药公司</td><td></td><td></td></tr><tr><td>聚维酮碘棉棒</td><td> Dynarex公司</td><td></td><td></td></tr><tr><td>聚丙烯缝线大小为6-0，3/8圈反向切割针</td><td> MYCO医疗</td><td></td><td></td></tr><tr><td> PE20管材</td><td> Becton Dickinson公司</td><td></td><td></td></tr><tr><td> 15微米的深红色的聚苯乙烯fluorospheres </td><td> Invitrogen公司</td><td></td><td></td></tr><tr><td> 1毫升汉密尔顿玻璃注射器</td><td>汉密尔顿公司</td><td> &lt;/ TD&gt; <td></td></tr><tr><td> 设备： </td><td></td><td></td><td></td></tr><tr><td>精灵加注射泵</td><td>肯特科学</td><td></td><td></td></tr><tr><td>荧光分光光度计</td><td> Digilab </td><td></td><td></td></tr><tr><td>啮齿动物呼吸机型号683 </td><td>哈佛设备</td><td></td><td></td></tr><tr><td></td><td></td><td></td><td> 表1表的具体的试剂和设备。 </td></tr></tbody></table>

angiogenesis in the ischemic rat lung

成人肺灌注全身支气管动脉和整个下肢静脉回流流经肺动脉。在大多数肺癌病症，它是规模较小的全身血管，增强肺灌注需要作出响应，并显示强大的新生血管。已经显示出肺血管局部缺血诱导的肺动脉梗阻导致快速全身动脉血管生成的人以及在几个动物模型。虽然组织学评估过程中的时间支气管动脉增殖的影响进行了仔细的描述的1的威贝尔，机制，负责本组织生长的新船是不明确的。我们提供左肺动脉缺血诱导的大鼠，导致支气管新生血管的手术细节。血管生成的程度的定量的呈现的两个内的肺血管床的存在下，由于一个额外的挑战。方法确定新生血管形成的基础上标记的微球注射。

血管铸型：左肺动脉缺血大鼠的影响的结果是在图1中所示。显示的是甲基丙烯酸酯演员的支气管血管和广泛的血管，气道左边的树后的28天内LPAL。要获得此铸造，注射全身血管与甲基丙烯酸酯的混合物（红色），降主动脉逆行进入气管插管和注射一个白色的基于硅的材料。血管铸型提供了一个显着的肺支气管血管生成的可视化。特别值得一提的是大，曲折上游的支气管血管的左肺。?...

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Angiogenesis in the Ischemic Rat Lung

肺灌注全身支气管动脉和肺动脉。在大多数肺病理学，它是较小的全身血管，显示了强大的新血管形成。停止肺血流量，促进轻快的支气管血管生成。我们提供诱导左肺动脉缺血，促进支气管新生血管的手术细节。

在缺血大鼠肺的血管生成

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

15.5K 次观看.  Johns Hopkins University. 肺灌注全身支气管动脉和肺动脉。在大多数肺病理学，它是较小的全身血管，显示了强大的新血管形成。停止肺血流量，促进轻快的支气管血管生成。我们提供诱导左肺动脉缺血，促进支气管新生血管的手术细节。

视频: Angiogenesis in the Ischemic Rat Lung

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.

利用磁共振成像，容积电脑断层扫描及超音波的裸鼠乳腺癌骨转移大鼠模型血管生成的多模态成像

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

科研

医学

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.

视网膜脱离模型鼠害由透明质酸钠的视网膜下注射

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.

纤维蛋白胶对小鼠肺移植研究肺特异性血管生成

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.

的隔离方法<em&gt;离体</em&gt;肺灌注的大鼠模型：教训制定鼠EVLP课程中学

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.

光化学诱导后部缺血性视神经病变的大鼠模型

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.

大鼠异位心脏腹部/在卷装配置单肺移植

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.

在大鼠缺血性皮肤创伤模型的演示

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

In Vitro Model of Coronary Angiogenesis

Here, we describe an in vitro culture assay to study coronary angiogenesis. Coronary vessels feed the heart muscle and are of clinical importance. Defects in these vessels represent severe health risks such as in atherosclerosis, which can lead to myocardial infarctions and heart failures in patients. Consequently, coronary artery disease is one of the leading causes of death worldwide. Despite its clinical importance, relatively little progress has been made on how to regenerate damaged coronary arteries. Nevertheless, recent progress has been made in understanding the cellular origin and differentiation pathways of coronary vessel development. The advent of tools and technologies that allow researchers to fluorescently label progenitor cells, follow their fate, and visualize progenies in vivo have been instrumental in understanding coronary vessel development. In vivo studies are valuable, but have limitations in terms of speed, accessibility, and flexibility in experimental design. Alternatively, accurate in vitro models of coronary angiogenesis can circumvent these limitations and allow researchers to interrogate important biological questions with speed and flexibility. The lack of appropriate in vitro model systems may have hindered the progress in understanding the cellular and molecular mechanisms of coronary vessel growth. Here, we describe an in vitro culture system to grow coronary vessels from the sinus venosus (SV) and endocardium (Endo), the two progenitor tissues from which many of the coronary vessels arise. We also confirmed that the cultures accurately recapitulate some of the known in vivo mechanisms. For instance, we show that the angiogenic sprouts in culture from SV downregulate COUP-TFII expression similar to what is observed in vivo. In addition, we show that VEGF-A, a well-known angiogenic factor in vivo, robustly stimulates angiogenesis from both the SV and Endo cultures. Collectively, we have devised an accurate in vitro culture model to study coronary angiogenesis.

In vitro models of coronary angiogenesis can be utilized for the discovery of the cellular and molecular mechanisms of coronary angiogenesis. In vitro explant cultures of sinus venosus and endocardium tissues show robust growth in response to VEGF-A and display a similar pattern of COUP-TFII expression as in vivo.

冠状血管生成体模型

Here, we describe an in vitro culture assay to study coronary angiogenesis. Coronary vessels feed the heart muscle and are of clinical importance. Defects ...

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.

温缺血/再灌注损伤的大鼠肺移植模型：改善结局的优化

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

<meta charset="utf8"></head><body>小鼠肺移植中细胞动力学的实时监测

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>作者聚焦：通过活体成像推进肺移植免疫学

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.

移植小鼠肺的活体双光子显微镜检查

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