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：動物use文：本研究におけるすべての手順を承認し、シンシナティ小児病院での動物実験使用とケア委員会（IACUC）のガイドラインに従って行った。 8週齢のC57BL / 6J雄マウスをジャクソン·ラボラトリーズ（バーハーバー、ME）から入手し、使用前に一週間順応させた。手術まで、動物を、無病原体バリア施設で飼育し、オートクレーブした食餌及びろ過水広告libdiumを与えた。各マウスのケージは、専用の空気と水を供給し、客室は、12時間の昼夜サイクルで維持した。手術からの回復に続いて、マウスは、濾過トップとのケージに維持し、オートクレーブ処理飼料適宜該ギプスを提供し、水のボトルからの水を濾過して設けられた。 楽器の調製<ol><li>ペーパークリップやピンを使用して6スキンリトラクターを作る。ツイストがl、紙·ピンのシャンク上のペーパークリップを伸ばした一端の5センチメートルストレート鋼線を庇ワイヤの端部にフック状の単0.5センチメートル &quot;U&quot;を作る。 </li><li>プラスチック製のラップを使用して、いくつかの15×15四方の手術用ドレープを作る。マウス当たり1ドレッシングを準備します。各ラップの間に紙塔を置く。 </li><li> 12×12インチのコル​​クタイル、ガーゼ、および綿棒のスタックと一緒にすべての手術器具を滅菌する。 </li></ol> 2.マウスの準備<ol><li> 2％イソフルランで麻酔を誘導する。動物を計量。 </li><li>専用の外科準備領域では、電気シェーバーと左胸部と首の領域を剃る。 </li><li>マウスの目に人工涙軟膏の低下を適用します。 </li><li>クロルヘキシジン、イソプロピルアルコールで首と左胸部を除染。さらに2回繰り返します。 </li></ol> 3.マウスオロ気管挿管および機械的人工換気<ol><li>予め温めておいた手術領域における非滅菌外科技術者の場所でマウス仰臥位を持っている。</li><li>足のピンチに応答がないことを文書化することにより、麻酔の深さを確認してください。 </li><li>手を洗うと手術衣装、マスク、帽子を着用した後、無菌手術用手袋を着用。 </li><li>ドレープと無菌技術を使用した後、喉頭を露出させ、前方半ば首オーバー1センチメートル垂直切開する。軽く湾曲し、鋸歯状の10cmの鉗子でストラップの筋肉を撤回し、ストレートハサミの先端でストラップの筋肉を広げることによって喉頭と気管を公開。 </li><li>経口半ば気管（ 図1A）に22 G鈍先端の血管カテーテルを挿入し、視覚的に配置します（ 図1B）を確認します。麻酔を維持し、げっ歯類の人工呼吸器を介して1〜3％イソフルランを使用して換気（;毎分200ストークスストロークあたり225μL）。 15cmのH 2 Oの圧力限界を採用</li></ol> 4.マウスの肺<ol><li>マウスの背中を向けた状態で、右側臥位でマウスを置き演算子（左側上）。無菌ドレープのように自己シールプラスチックラップを使用してください。ドレープを切断、4 番目と5 番目の肋間で肋骨に2cmの長さのカット並列を作るために鈍い傾いて湾曲したハサミを使用しています。鈍先端湾曲したハサミを挿入し、根底にある肋骨と肋間筋から離れて皮膚を解剖。 </li><li>正方形の1.5×1.5センチメートル外科ウィンドウ（ 図2A）を露出するために4つのリトラクターで皮膚を撤回。コルクボードにリトラクターを固定します。 </li><li>湾曲した鉗子を用いてリブまで細かく分析し、胸腔を入力するように湾曲した鉗子の1チップを使用しています。 </li><li>丸い先端マイクロはさみを使用して、胸腔を入力するように下刃を使用しています。反対方向にリブを繰り返し間で0.5cmの切開を行います。 </li><li>残りの2つのリトラクターを使用して、前後軸に胸部を開き、コルクボード（ 図2B）に開創を確保。 </li> &lt;LI&gt;左肺をつかみ、左肺動脈と気管支が（ 図3A、B）が露出するまで開胸を通って横方向及び下方に左肺の上部を変位、左手に湾曲した鈍先端が鉗子を使用。 </li><li>手のひらでアプリケーターとパーム（ 図3C）からポインティング曲がった先端の本体と右手にロードされたチタン血管をマイクロアプリケーターを持ち、左の後面の曲率に沿って胸部にアプリケータ先端部をスライドさせ左気管支及び肺動脈（ 図3D）の肺およびクリップ。 </li><li>アプリケーターを取り外しますが後退し左肺を保つ。右手で鈍先端マイクロハサミを握り、クリップに気管支及び肺動脈遠位をカットし、左肺（ 図3E）を削除。 </li><li>リブリトラクターを削除します。 </li><li> incisiに劣る皮膚から1cmを挟まないように湾曲した鈍鉗子を使用してくださいではなく、振動板のレベルを超えて、皮膚を通して、左胸腔（ 図4A、B）への24のG血管カテーテルを挿入します。 </li><li>胸腔を閉じるには、4 番目と5 番目の肋骨の周りに2結節縫合を配置する5-0プロレン縫合糸を使用してください。 </li><li>肌のリトラクターを削除します。切開の長さに沿って皮膚を近似し、肌が閉じ接着する鉗子の二組を使用してください。 </li><li>血管カテーテルに3ミリリットルルアーロックシリンジを接続し、穏やかな吸引を適用し、血管カテーテルを引き出すことにより、残留空気を除去する。 </li><li>首の切開が以前鉗子の二組を使用して閉じた接着。 </li></ol> 5.マウスシャム肺全摘術<ol><li> 「マウス肺」プロトコルに記載されているように左肺を公開します。左胸腔内に空気を可能にするために湾曲した鈍鉗子を用いて胸郭を持ち上げて（ 図5A、B）。 </li><li>左カントーに24のG血管カテーテルを配置左肺を傷つけないように注意しながら、上記のようにracic空洞。 </li><li> 5-0プロレン縫合糸を使用し、肺（ 図5C）を穿刺しないように注意しながら、 第 3 / 第 4に縫合材料の2つの長さと5 番目 / 6 番目の肋骨隙間（ 図5D）を配置。左肺ヘルニアのリスクを軽減するために抱き合わせ前に縫合材料の両方の長さを置きます。 2、中断ステッチ（ 図5E）を作るために縫合材料を結ぶ。 </li><li>胸部切開上の皮膚を接着、血管カテーテルで残留空気を除去し、上記のように首の切開を接着。 </li></ol> 6.蘇生、鎮痛、および回復<ol><li>イソフルランの電源を切り、ブプレノルフィンの0.1 mg / kgを投与し、生理食塩水を皮下の0.5ミリリットル。 </li><li>自発呼吸が再開すると、気管内チューブを取り外します。 </li><li>それが再び歩行になるまでマウスを観察します。通常resumウォーキング気管内チューブを除去した後、数分、ES。 </li><li> O / Nを回復するために27℃のインキュベーター（加湿され、25％の酸素）でマウスを置きます。 注：我々は、手術後の最初の24時間、ケージの床に水で湿らせた固形飼料のいくつかのペレットを配置。 </li><li>手術後3日間、一日二回腹腔内注射によりブプレノルフィンの0.1 mg / kgをを管理します。動物を扱うときに手術部位をオープンしないように注意してください。 </li></ol> 7.マウスの監視<ol><li>手術後1、3、5、および7日目にマウスを秤量する。 </li></ol>

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The authors have nothing to disclose.

我々はこれまでに報告マウスPNXとマウス偽PNX手順のほとんどの詳細な説明を提供してきました。私たちは、手順を学ぶ研究者は、一般的に遭遇する、と​​我々はこれらの落とし穴を軽減するために私たちの研究室が開発したいくつかの技術を概説している共通の落とし穴のいくつかの読者は意識してきた。このモデルを利用する他の研究所は、他の技術の変更を開発したか、別の商品を利?...

肺の主な機能は、生物と大気の酸素と二酸化炭素の交換を提供することである。ヒトでは、先天性および後天条件のホストが損なわれた肺機能をもたらす減少、肺の表面積をも​​たらす。 すなわち、肺-そのような吸入コルチコステロイド、気管支拡張薬、酸素補給、および慢性の機械的換気などの治療のホストが損なわれた肺機能1-3の影響を緩和するために使用されているが、これらの状態のための理想的な治療は、機能的な肺組織の再生を促進する再生。 哺乳動物の組織再生は十分に立証されている。アフリカのトゲマウスは瘢痕形成4なく、皮膚の大部分を再生することができる。ヒトでの末節骨は、以下の傷害または切断5-7を再生成することができます。以下の肺（PNX）、代償肺の成長が行い、9ラット、マウス8で発生gsは10、およびヒト11。定義により、代償肺の成長は既存の空域の拡大が、関連した微小循環12の拡大にこれらの拡大空域の再隔壁形成だけではないが含まれます。遺伝子発現解析は、このモデルが肺の発達13のシグナル伝達事象の多くを再現することを示した。四週間マウスPNXた後、肺胞の表面積は、偽手術動物14と同等である。本稿では、我々は、マウスPNXおよび偽PNX手順を説明します。

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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と偽肺の両方のための100％ - PNXと偽手術マウスの体重のプロットを図6に提供されている我々の手では、生存率は一貫して95です。右肺がこのモデルと予想される時間経過に再成長する方法の詳細については、我々はギブニーら 15及びWang らの原稿を読者に参照してください。14 いくつかの一般的な落とし穴が正常マウ...

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.

代償肺の成長のマウス肺モデル

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 ビュー.  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.

ビデオ: 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.

同所性肺移植のマウスモデルにおける閉塞性細気管支炎の開発

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

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.

マウス肺切除術および義歯移植による内臓肺表面積の標準化法

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

発生生物学

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.

マウス肺における圧力 - 容積曲線の測定

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.

繰り返しオゾン暴露によるマウスの慢性閉塞性肺疾患モデルの生成

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.

組み合わせてモノクロまたは Sugen 肺高血圧ラットのモデルとして左肺全摘

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.

肺炎の治安を評価するための実験モデル

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

免疫・感染学

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.

温虚血/再灌流損傷のラット肺移植モデル:転帰を改善するための最適化

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.

正常母性陰圧換気 ex situ 肺灌流:肺機能と代謝の評価

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.