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.

注:动物使用声明:在本研究中的所有程序均经批准，并按照机构动物使用及护理委员会（IACUC）在辛辛那提儿童医院的指导方针进行。八周龄C57BL / 6J雄性小鼠从Jackson实验室（巴港，ME）中获得，并使其适应环境，使用前一个星期。直到手术，将动物饲养在无病原体的阻挡设施，并提供高压灭菌食物和过滤水的广告libdium。每个鼠笼用专用的空气和水的供给，并提供维持在12小时昼夜周期。以下从手术中恢复，小鼠保持在笼子里与过滤上衣，提供蒸压州城广告libidum，并提供了从一瓶水过滤水。 1.准备仪器<ol><li>使用曲别针和大头针6皮肤拉钩。扭曲拉直回形针在纸张的针柄，升屋檐5cm的直钢丝的一端，使单个0.5厘米"U"形钩在导线的末端。 </li><li>使用保鲜膜约15×15厘米见方的手术单。准备每只小鼠1敷料。把一纸塔在每个包装之间。 </li><li>消毒所有的手术工具以及一叠12×12英寸的软木砖，纱布，棉花拭子。 </li></ol> 2.鼠标准备<ol><li>诱导麻醉，用2％异氟醚。称量动物。 </li><li>在一个专门的手术准备区刮胡子左胸部和颈部电气剃须刀。 </li><li>申请一滴人工泪液软膏鼠标的眼睛。 </li><li>净化颈部和左胸部与洗必泰和异丙醇。重复两次以上。 </li></ol> 3.鼠标奥罗气管插管和机械通气<ol><li>有一个非无菌手术技术人员代替鼠标仰卧在预热手术区。</li><li>通过记录缺乏爪子掐的响应确认麻醉深度。 </li><li>洗手，并穿上手术装束，口罩，帽子后，穿上无菌手术手套。 </li><li>立体裁剪，并使用无菌技术后，使超过前中期的脖子，露出喉部有1厘米的垂直切口。轻轻缩回带状肌与弯曲的，锯齿10厘米钳，并通过与一个直剪刀的前端扩展的带状肌暴露喉和气管。 </li><li>口服插入一个22克钝尖血管导管插入中期气管（ 图1A）和目视确认放置（ 图1B）。维持麻醉和通风用1-3％异氟醚通过啮齿动物呼吸机（225微升每冲程;每分钟200斯托克斯）。采用15厘米H 2 O的压力极限</li></ol> 4.鼠标肺切除<ol><li>莱鼠标在右侧卧位，用鼠标的背面朝上操作员（左侧上）。用自密封的保鲜膜作为无菌的悬垂性。切割通过悬垂性好，用钝尖剪子弯，使长切割平行于肋骨2厘米，在第 4和第 5肋间。将钝头剪刀弯曲和底层的肋骨和肋间肌肉解剖皮肤了。 </li><li>缩回四个拉钩皮肤暴露正方形1.5×1.5cm的外科窗口（ 图2A）。固定拉钩，以软木板上。 </li><li>解剖下来使用弯钳肋，并使用弯钳之一尖端进入胸腔。 </li><li>使用钝头微型剪刀，使用下刀片进入胸腔。使肋和重复在相反的方向之间为0.5厘米的切口。 </li><li>使用剩下的两个拉钩，在前后轴和安全的拉钩到软木板上（ 图2B）打开胸腔。 </li> &lt;李&gt;使用在左手弯曲钝尖镊子，把握左肺和置换左肺的上部横向和下方通过开胸直到左肺动脉与支气管露出（ 图3A，B）。 </li><li>持在右手装入钛血管microclip敷贴与涂抹在手掌和弯曲尖端从手掌（ 图3C）背向的主体中，沿左侧的后方面的曲率滑动施用尖端进入胸腔肺和剪辑左支气管和肺动脉（ 图3D）。 </li><li>取下敷贴但保留左肺缩回。抓住钝尖微型剪刀用右手切支气管和肺动脉远端剪辑，删除左肺（ 图3E）。 </li><li>拆下肋骨拉钩。 </li><li>使用弯曲钝钳捏起来1厘米皮肤逊色于incisi上，但上述隔膜的水平，并插入穿过皮肤并进入左胸腔（ 图4A，B）的一个24克血管导管。 </li><li>使用5-0 prolene线缝合周围放置第 4 次 2间断缝合和第 5肋关闭胸腔。 </li><li>去除皮肤拉钩。使用两套镊子来近似沿切口的长度皮肤和粘在皮肤闭合。 </li><li>一个3毫升路厄锁注射器连接到血管导管和通过施加轻柔抽吸抽出血管导管除去残留的空气。 </li><li>胶颈部切口闭合使用镊子两套像以前一样。 </li></ol> 5.鼠标深水肺切除<ol><li>暴露左肺如"老鼠肺切除"协议指出。提起肋笼具有弯曲钝钳，以允许空气进入左胸腔（ 图5A，B）。 </li><li>放置一个24克血管导管插入左寿拉契奇腔上面，小心不要弄伤左肺。 </li><li>使用5-0普理灵缝合小心不要刺破肺（ 图5C），将两个长度的缝合材料到3 次 / 4和第 5/6 次肋骨间隙（ 图5D）。绑减轻左肺症的风险之前，将缝合材料的两个长度。领带的缝合材料，以使2中断线圈（ 图5E）。 </li><li>胶上的皮肤胸切开，除去残留的空气与血管导管，和胶水颈部切口如上。 </li></ol> 6.复苏，镇痛和恢复<ol><li>关掉异氟烷，并管理0.1毫克/公斤丁丙诺啡和0.5生理盐水皮下注射毫升。 </li><li>当自主呼吸恢复，取出气管插管。 </li><li>观察鼠标，直到它再次走动。通常走resum上课除去气管导管后几分钟。 </li><li>放置在27℃培养箱（加湿，25％氧气）鼠标恢复O / N。 注意:我们放置食物与水润湿的上箱地板为手术后的第一个24小时的几个粒料。 </li><li>手术后每日三天两次施用0.1毫克/千克丁丙诺啡腹腔注射。小心不要处理的动物时，打开手术部位。 </li></ol> 7.鼠标监测<ol><li>手术后称量小鼠在1，3,5和7天。 </li></ol>

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The authors have nothing to disclose.

我们已经提供了迄今报告的鼠标PNX和鼠标假PNX程序的最详细的描述。我们已经取得的读者了解一些常见的陷阱，调查人员学习的过程通常会遇到，我们已经介绍了我们的实验室开发，以减轻对这些缺陷的几个技巧。利用这种模式其他实验室可能已经开发了其他的技术修改或使用不同的仪器。在评估的技术差异，个别研究者将需要决定是否这些差异显著影响实验结果。 补偿性?...

肺的主要功能是提供一个有机体和大气之间的氧气和二氧化碳交换。在人类中，一台主机的先天和后天条件导致降低的肺表面面积导致肺功能受损。虽然宿主疗法，如吸入皮质类固醇，支气管扩张剂，补充氧气和慢性机械通气用于减轻肺功能受损1-3的后果，提供了理想的治疗，这些条件将促进功能的肺组织的再生- 即，肺再生。 哺乳动物组织再生已经有据可查。非洲刺鼠可再生大面积皮肤无疤痕形成4。在人类的末节指骨可再生损伤后或截肢5-7。继肺（PNX），肺代偿出现增长小鼠8只9，做GS 10和人类11。根据定义，补偿性肺生长包括现有的空域与扩展相关的微循环12不仅扩大这些扩大空域，但重新septation。基因表达分析表明，这种模式概括很多肺发育13的信号事件。四周后小鼠PNX，肺泡表面面积相当于假手术的动物14。在这个手稿中，我们描述鼠标PNX和假PNX程序。

<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.

。PNX，假手术鼠标重量的情节在图6提供了在我们的手中，生存始终是95 - 100％为PNX和假肺。有关如何右肺再生长在该模型和预期的时间过程中，我们建议读者参考吉布尼等人的 15和Wang 等人的手稿的描述。14 几种常见的陷阱，必须避免成功执行鼠标PNX和鼠标假肺的程序。 气管插管 :许多研究者学习?...

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.

补偿性增长龙鼠标肺切除型号

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

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

12.8K 次观看.  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.

视频: 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.

闭塞性细支气管炎在原位肺移植小鼠模型的发展

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

科研

医学

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.

通过小鼠肺切除术和假体植入测量内部肺表面积的标准方法

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

发育生物学

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.

在小鼠肺压力容积曲线的测量

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.

反复臭氧暴露对小鼠慢性阻塞性肺疾病模型的产生

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.

左肺切除术联合单氯联氨酸或苏根为大鼠肺动脉高压的模型

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.

评估肺炎消退的实验模型

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

免疫与感染

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 ...

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.

非原位负压通气肺灌注:肺功能和代谢的评估

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.

Ex Vivo Porcine Experimental Model for Studying and Teaching Lung Mechanics

We present an ex vivo pig lung model for the demonstration of pulmonary mechanics and alveolar recruitment maneuvers for teaching purposes. The lungs can be used for more than one day (up to five days) with minimal changes in pulmonary mechanics variables.

离体 猪肺力学研究与教学的实验模型

Mechanical ventilation is widely used and requires specific knowledge for understanding and management. Health professionals in this field may feel ...

补偿性增长龙鼠标肺切除型号

摘要

探索更多视频

此视频中的章节

闭塞性细支气管炎在原位肺移植小鼠模型的发展

通过小鼠肺切除术和假体植入测量内部肺表面积的标准方法

在小鼠肺压力容积曲线的测量

反复臭氧暴露对小鼠慢性阻塞性肺疾病模型的产生

左肺切除术联合单氯联氨酸或苏根为大鼠肺动脉高压的模型

评估肺炎消退的实验模型

温缺血/再灌注损伤的大鼠肺移植模型:改善结局的优化

非原位负压通气肺灌注:肺功能和代谢的评估

A Simple Cuff Technique for Murine Left Lung Transplantation

离体猪肺力学研究与教学的实验模型

补偿性增长龙鼠标肺切除型号

摘要

探索更多视频

此视频中的章节

闭塞性细支气管炎在原位肺移植小鼠模型的发展

通过小鼠肺切除术和假体植入测量内部肺表面积的标准方法

在小鼠肺压力容积曲线的测量

反复臭氧暴露对小鼠慢性阻塞性肺疾病模型的产生

左肺切除术联合单氯联氨酸或苏根为大鼠肺动脉高压的模型

评估肺炎消退的实验模型

温缺血/再灌注损伤的大鼠肺移植模型:改善结局的优化

非原位负压通气肺灌注:肺功能和代谢的评估

A Simple Cuff Technique for Murine Left Lung Transplantation

离体 猪肺力学研究与教学的实验模型

离体猪肺力学研究与教学的实验模型