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.

HINWEIS: Tier Verwendung Aussage: Alle Verfahren in dieser Studie wurden mit Genehmigung und nach den Richtlinien der Institutional Animal Benutzung und Pflege Committee (IACUC) in Cincinnati Kinderklinik durchgeführt. Acht Wochen alte C57BL / 6J männliche Mäuse wurden von Jackson Laboratories (Bar Harbor, ME) erhalten und man ließ sie für eine Woche vor dem Gebrauch akklimatisieren. Bis der Operation wurden die Tiere in einem pathogenfreien Barriere Anlage untergebracht und zur Verfügung gestellt autoklaviert Chow und gefiltertes Wasser ad libdium. Jede Maus Käfig wurde mit einer dedizierten Luft und Wasser versorgt, und die Zimmer wurden auf einem 12 Stunden Tag-Nacht-Zyklus gehalten. Nach Erholung von der Operation wurden die Mäuse in Käfigen mit gefiltertem Spitzen aufrechterhalten, sofern autoklaviert Futter ad libidum und sofern aus einer Wasserflasche gefiltertes Wasser. 1. Herstellung des Instruments <ol><li> Stellen Sie 6 Haut Wundhaken mit Büroklammern und Pins. Twist bogene Büroklammern auf den Schäften der Papier-Pins, lTraufe eine 5 cm gerader Stahldraht an einem Ende und machen eine einzige 0,5 cm &quot;U&quot; förmigen Haken am Ende des Drahtes. </li><li> Stellen ca. 15 x 15 cm im Quadrat-OP-Abdeckungen mit Plastikfolie. Bereiten Sie eine Umkleide pro Maus. Setzen Sie ein Papierturm zwischen den einzelnen Wickel. </li><li> Sterilisieren Sie alle chirurgische Instrumente zusammen mit einem Stapel von 12 x 12 Zoll Korkplatten, Gaze, und Wattestäbchen. </li></ol> 2. Maus Vorbereitung <ol><li> Induzieren Anästhesie mit 2% Isofluran. Wiegen Tier. </li><li> In einem speziellen Operationsvorbereitungsbereich rasieren linken Brust und Nackenbereich mit elektrischen Rasierer. </li><li> Geben Sie einen Tropfen der künstliche Tränen Salbe die Augen die Maus. </li><li> Dekontaminieren Hals und linken Brust mit Chlorhexidin und Isopropylalkohol. Wiederholen Sie zweimal. </li></ol> 3. Maus-Oro-Intubation und Beatmung <ol><li> Haben Sie ein nicht-sterilen OP-Techniker statt mit der Maus auf dem Rücken in der vorgewärmten OP-Bereich.</li><li> Narkosetiefe Bestätigen Dokumentation Fehlen einer Reaktion auf die Pfote Prise. </li><li> Nach dem Waschen der Hände und das Anziehen OP-Kleidung, Maske und Hut, don sterile Handschuhe. </li><li> Nach Drapierungen und unter aseptischen Bedingungen, machen Sie einen 1 cm vertikale Inzision über der vorderen Mitte Nacken, den Kehlkopf freizulegen. Leicht einfahren Halsmuskulatur mit gebogenen, gezackten 10 cm Pinzette und setzen den Kehlkopf und Luftröhre durch die Streuung der Halsmuskulatur mit der Spitze einer geraden Schere. </li><li> Oral legen Sie eine 22 G stumpfen Spitze Angiokatheter in die Mitte der Luftröhre (1A) und optisch Platzierung (1B) zu bestätigen. Pflegen Anästhesie und belüften mit 1-3% Isofluran durch Nagetierventilator (225 ul pro Hub; 200 Stokes pro min). Verwenden Sie ein Druckgrenze von 15 cm H 2 O </li></ol> 4. Maus Pneumonektomie <ol><li> Legen Sie die Maus in die richtige Seitenlage mit der Maus den Rücken vorder Betreiber (links oben). Verwenden Sie einen selbstdichtenden Plastikfolie als sterilen Tuch. Schneiden durch den Faltenwurf, verwenden stumpfer Spitze gebogene Schere, einen 2 cm langen Schnitt parallel zu den Rippen auf der 4. und 5. Zwischenrippenraum zu machen. Legen Sie die stumpfe Spitze gebogene Schere und sezieren die Haut von den zugrunde liegenden Rippen und Rippenmuskeln. </li><li> Ziehen Sie die Haut mit vier Wundhaken, um ein Quadrat aussetzen 1,5 x 1,5 cm Operationsfenster (Abbildung 2A). Sichern Sie die Wundhaken an den Kork Bord. </li><li> Präparieren Sie auf Rippen mit einer gebogenen Pinzette, und verwenden Sie eine Spitze der gebogenen Pinzette, um die Brusthöhle zu gelangen. </li><li> Mit Hilfe der stumpfen Spitze Mikroscheren, verwenden Sie die Untermesser, um die Brusthöhle zu gelangen. Machen Sie eine 0,5 cm Einschnitt zwischen den Rippen, und wiederholen Sie in die entgegengesetzte Richtung. </li><li> Mit den beiden verbleibenden Wundhaken, öffnen Sie den Brustkorb in der Vorwärts-Rückwärts-Achse und die sichere die Retraktoren mit dem Kork Bord (2B). </li> &lt;li&gt; Mit gebogenen stumpfen Pinzette in der linken Hand das linke Lunge und verdrängen den oberen Teil der linken Lunge seitlich und unten durch die Thorakotomie, bis die linke Lungenarterie und Bronchien ausgesetzt sind (3A, B). </li><li> Halten Sie die geladenen Titan Gefäßmikroclip Applikator in die rechte Hand mit dem Körper des Applikators in der Handfläche und gebogene Spitze nach weg von der Handfläche (3C), schieben Sie die Applikatorspitze in den Brustkorb entlang der Krümmung der hinteren Seite des linken Lunge und Clip der linken Bronchus und Lungenschlagader (3D). </li><li> Entfernen Sie den Applikator aber halten Sie die linke Lunge eingefahren. Fassen Sie die stumpfe Spitze Mikro-Schere mit der rechten Hand und schneiden Sie die Bronchien und Lungenarterie distal der Clip und entfernen linken Lunge (3E). </li><li> Entfernen Sie die Rippenspreizer. </li><li> Mit den gebogenen stumpfen Pinzette zu kneifen bis 1 cm von der Haut schlechter als die incisiauf, sondern über dem Niveau der Membran und setzen Sie eine 24 G Angiokatheter durch die Haut und in die linke Brusthöhle (4A, B). </li><li> Verwenden Sie 5-0 Prolene Naht zu zwei Knopfnähten auf der 4. und 5. Rippen platzieren, um die Brusthöhle zu schließen. </li><li> Entfernen Sie die Haut Wundhaken. Verwenden Sie zwei Sätze von Pinzette, um die Haut entlang der Länge des Einschnitts nähern und kleben die Haut verschlossen. </li><li> Schließen Sie einen 3 ml Luer-Lock-Spritze mit dem Angiokatheter und entfernen Restluft durch leichten Ansaugen und Zurückziehen des Angiokatheter. </li><li> Kleben Sie den Halsschnitt geschlossen mit zwei Sätzen von Zangen wie zuvor. </li></ol> 5. Maus Sham Pneumonektomie <ol><li> Setzen Sie die linke Lunge wie in &quot;Maus Pneumonektomie&quot; Protokoll vermerkt. Heben Sie den Brustkorb mit gebogenen stumpfen Pinzette, um Luft in die linke Brusthöhle zu ermöglichen (5A, B). </li><li> Legen Sie eine 24 G Angiokatheter in die linke thoRacic Hohlraum wie oben darauf achten, daß die linke Lunge schädigen. </li><li> Mit 5-0 Prolene Nahtmaterial und darauf achten, daß die Lunge (5C) durchstoßen, indem Sie zwei Längen von Nahtmaterial in der 3. / 4 und 5. / 6. Rippenzwischenräumen (5D). Gebe beide Längen von Nahtmaterial vor den Bund, um die Risiken der linken Lunge Herniation zu verringern. Binden Sie die Nahtmaterial zu zwei unterbrochenen Stichen (5E) zu machen. </li><li> Kleben Sie die Haut über der Brust-Schnitt, entfernen Sie Restluft mit dem Angiokatheter und kleben Sie den Halsschnitt wie oben. </li></ol> 6. Wiederbelebung, Analgesie und Wiederherstellung <ol><li> Schalten Sie die Isofluran und Verwalten 0,1 mg / kg von Buprenorphin und 0,5 ml physiologischer Kochsalzlösung subkutan. </li><li> Wenn spontane Atmung wieder aufzunehmen, entfernen Sie den Endotrachealtubus. </li><li> Beachten Maus, bis es wieder ambulante ist. Wandern in der Regel Resum¯es mehrere Minuten nach der Entfernung des Endotrachealtubus. </li><li> Platzieren Sie die Maus in einem 27 ° C Inkubator (befeuchteten, Sauerstoff 25%) zu erholen O / N. HINWEIS: Legen Sie mehrere Pellets von Chow mit Wasser für die ersten 24 Stunden nach der Operation benetzt auf dem Käfigboden. </li><li> Verabreichen 0,1 mg / kg Buprenorphin durch intraperitoneale Injektion zweimal täglich für drei Tage nach der Operation. Darauf achten, dass die Operationsstelle beim Umgang mit Tieren zu öffnen. </li></ol> 7. Maus-Überwachung <ol><li> Wiegen Mäusen nach 1, 3, 5 und 7 Tage nach der Operation. </li></ol>

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The authors have nothing to disclose.

Wir haben die detaillierte Beschreibung der Maus PNX und Maus Schein PNX Verfahren bis heute berichtet vorgesehen. Wir haben die Leser bewusst einige der häufigsten Fallstricke, dass die Ermittler das Erlernen der häufigsten Verfahren stoßen gemacht, und wir haben verschiedene Techniken, die von unserem Labor entwickelt, um gegen diese Gefahren zu mildern skizziert. Andere Labors nutzen dieses Modell können andere Technik Modifikationen entwickelt haben, oder verwenden Sie verschiedene Instrumente. Bei der Bewertung...

Die Hauptfunktion der Lunge ist für Sauerstoff und Kohlendioxid Austausch zwischen einem Organismus und der Atmosphäre bereitzustellen. Beim Menschen eine Vielzahl von angeborenen und erworbenen Bedingungen führen zu verminderter Lungenfläche, die zu einer Beeinträchtigung der Lungenfunktion führt. Dh Lungen - Obwohl eine Vielzahl von Therapien wie inhalativen Kortikosteroiden, Bronchodilatatoren, zusätzlichen Sauerstoff und chronische mechanische Belüftung verwendet werden, um die Folgen der Beeinträchtigung der Lungenfunktion 1-3 abzuschwächen, wäre die ideale Therapie für diesen Bedingungen Nachwachsen der funktionellen Lungengewebe zu fördern Regeneration. Säugetiergeweberegeneration ist gut dokumentiert worden. Die Afrikanische Stachelmaus können große Bereiche der Haut ohne Narbenbildung 4 zu regenerieren. Die Phalanx beim Menschen nach einer Verletzung oder Amputation 5-7 regenerieren. Nach Pneumonektomie (PNX) tritt Ausgleichslungenwachstum bei Mäusen, 8, 9 Ratten, zu tungs 10 und 11 Menschen. Per Definition beinhaltet Ausgleichslungenwachstum nicht nur Ausbau der bestehenden Lufträume, aber erneut Septierung dieser vergrößerten Lufträume mit der Expansion des zugehörigen Mikrozirkulation 12. Genexpressionsanalyse hat gezeigt, dass dieses Modell rekapituliert viele der Signalwege der Lungenentwicklung 13. Vier Wochen nach der Maus PNX ist Alveolaroberfläche äquivalent zu der scheinoperierten Tieren 14. In diesem Manuskript beschreiben wir die Maus PNX und Schein PNX Verfahren.

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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.

. Ein Grundstück von PNX und scheinoperierten Maus Gewichte ist in Abbildung 6 vorgesehen In unseren Händen, ist das Überleben konsequent 95-100% für beide PNX und Schein Pneumonektomie. Eine Beschreibung, wie die rechte Lunge wieder wächst in diesem Modell und dem erwarteten zeitlichen Verlauf, verweisen wir den Leser auf Manuskripte Gibney et al. 15 und Wang et al. 14 Einige häufige Fehler zu vermeiden, um die Maus PNX und Maus...

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.

Maus Pneumonektomie Modell der Ausgleichs Lung Wachstum

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 Aufrufe.  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.

Serie: 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.

Entwicklung einer Bronchiolitis obliterans in einem Mausmodell der orthotopen Lungentransplantation

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

Forschung

Medizin

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.

Eine standardisierte Methode zur Messung der inneren Lungenoberfläche über Maus Pneumonektomie und Prothesenimplantation

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

Entwicklungsbiologie

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.

Die Messung der Druck-Volumen-Kurve in der Mäuselunge

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.

Generation von eine chronisch obstruktive Lungenerkrankung-Modell bei Mäusen durch wiederholte Ozonbelastung

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.

Die linke Pneumonektomie kombiniert mit Monocrotaline oder Sugen als Modell der pulmonalen Hypertonie bei Ratten

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.

Experimentelles Modell zur Evaluierung des Abklingens von Lungenentzündungen

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

Immunologie und Infektion

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.

Ein Ratten-Lungentransplantationsmodell für warme Ischämie/Reperfusionsschäden: Optimierungen zur Verbesserung der Ergebnisse

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.

Normotherme Unterdruckbeatmung ex situ Lungenperfusion: Beurteilung der Lungenfunktion und des Lungenstoffwechsels

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.

<meta charset="utf8"></head><body>Eine einfache Manschettentechnik für die Transplantation der linken Lunge von Mäusen

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.

Eine einfache Manschettentechnik für die Transplantation der linken Lunge von Mäusen

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

Maus Pneumonektomie Modell der Ausgleichs Lung Wachstum

Zusammenfassung

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Die Messung der Druck-Volumen-Kurve in der Mäuselunge

Generation von eine chronisch obstruktive Lungenerkrankung-Modell bei Mäusen durch wiederholte Ozonbelastung

Die linke Pneumonektomie kombiniert mit Monocrotaline oder Sugen als Modell der pulmonalen Hypertonie bei Ratten

Experimentelles Modell zur Evaluierung des Abklingens von Lungenentzündungen

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Eine einfache Manschettentechnik für die Transplantation der linken Lunge von Mäusen

Eine einfache Manschettentechnik für die Transplantation der linken Lunge von Mäusen