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.

Declaración de uso animal:: NOTA Todos los procedimientos de este estudio se llevaron a cabo con la aprobación y siguiendo las directrices de la Institutional Animal Uso y Comité de Cuidado (IACUC) en el Hospital de Niños de Cincinnati. C57BL / 6J macho de ocho semanas de edad se obtuvieron de Jackson Laboratories (Bar Harbor, ME) y se dejaron aclimatar durante una semana antes de usar. Hasta la cirugía, los animales fueron alojados en un centro de barrera libre de patógenos y siempre pienso en autoclave y se filtra agua ad libdium. Cada jaula ratón se suministra con un aire dedicado y agua, y las habitaciones se mantuvieron en un ciclo día-noche de 12 horas. Después de la recuperación de la cirugía, los ratones fueron mantenidos en jaulas con las tapas filtrados, siempre autoclave comida ad libitum, y siempre que filtran el agua de una botella de agua. 1. Elaboración de Instrumentos <ol><li> Hacer 6 retractores de la piel utilizando clips y pines de papel. Giro enderezó clips de papel en los vástagos de los pines de papel, lalero de un alambre de acero recta 5 cm en un extremo y hacer un solo 0.5 cm &quot;U&quot; en forma de gancho en el extremo del cable. </li><li> Hacer un poco de 15 x 15 cm paños quirúrgicos cuadrados utilizando una envoltura de plástico. Prepare un aderezo por ratón. Ponga una torre de papel entre cada envoltura. </li><li> Esterilizar todos los instrumentos quirúrgicos junto con una pila de fichas de 12 x 12 pulgadas de corcho, gasas, y aplicadores de algodón. </li></ol> 2. Preparación del ratón <ol><li> Inducir la anestesia con isoflurano al 2%. Pesar animal. </li><li> En un área de preparación quirúrgica dedicada afeitarse tórax izquierdo y el cuello con la máquina de afeitar eléctrica. </li><li> Aplique una gota del ungüento de lágrimas artificiales para los ojos del ratón. </li><li> Descontaminar el cuello y el tórax izquierdo con clorhexidina y alcohol isopropílico. Repite dos veces más. </li></ol> 3. Ratón Oro-traqueal La intubación y ventilación mecánica <ol><li> Haga que un técnico quirúrgico lugar no estéril la supina ratón en el área quirúrgica pre-calentado.</li><li> Confirme la profundidad de la anestesia mediante la documentación de la falta de una respuesta a la pizca de la pata. </li><li> Después de lavarse las manos y ponerse el traje quirúrgico, máscara y sombrero, ponerse guantes quirúrgicos estériles. </li><li> Después de drapeado y utilizando una técnica aséptica, hacer una incisión vertical de 1 cm en el mediano anterior del cuello para exponer la laringe. Ligeramente retraer los músculos de la correa dentada, con curvas de 10 cm pinzas y exponer la laringe y la tráquea mediante la difusión de los músculos de la correa con la punta de unas tijeras rectas. </li><li> Por vía oral insertar un 22 G angiocatéter romo-punta en la mitad de la tráquea (Figura 1A) y confirmar visualmente la colocación (Figura 1B). Mantener la anestesia y ventilar utilizando 1-3% isoflurano a través del ventilador de roedores (225 l por golpe; 200 Stokes por minuto). Emplear un límite de presión de 15 cm H 2 O. </li></ol> 4. Ratón neumonectomía <ol><li> Coloque el ratón en la posición de decúbito lateral derecho con la espalda del ratón haciael operador (a la izquierda hacia arriba). Utilice una envoltura de plástico de cierre automático como un campo estéril. Corte a través de la cortina, utilice contundentes tijeras curvas de punta para hacer un 2 cm de largo corte paralelo a las costillas en el espacio intercostal 4 º y 5 º. Inserte las tijeras de punta curvada contundentes y diseccionar la piel lejos de las costillas subyacentes y los músculos intercostales. </li><li> Retraer la piel con cuatro retractores para exponer un cuadrado 1,5 x 1,5 cm de la ventana quirúrgica (Figura 2A). Asegure los retractores al tablero de corcho. </li><li> Diseccionar a costillas utilizando pinzas curvas, y utilizar una punta de las pinzas curvas para entrar en la cavidad torácica. </li><li> Usando la punta roma micro-tijeras, utilice la cuchilla inferior para entrar en la cavidad torácica. Hacer una incisión de 0,5 cm entre las costillas y la repetición en la dirección opuesta. </li><li> Usando los dos retractores restantes, abrir el tórax en el eje antero-posterior y el seguro de los retractores de la placa de corcho (Figura 2B). </li> &lt;li&gt; Con unas pinzas de punta roma curvas en la mano izquierda, sujete el pulmón izquierdo y desplazar a la parte superior del pulmón izquierdo lateral e inferior a través de la toracotomía hasta la arteria pulmonar izquierda y bronquios están expuestos (Figura 3A, B). </li><li> Sosteniendo el titanio microclip vascular aplicador cargado en la mano derecha con el cuerpo del aplicador en la palma y punta curva en dirección opuesta a la palma de la mano (Figura 3C), deslice la punta del aplicador en el tórax a lo largo de la curvatura de la cara posterior de la izquierda pulmón y el clip del bronquio izquierdo y la arteria pulmonar (Figura 3D). </li><li> Retire el aplicador pero mantener el pulmón izquierdo retraída. Sujete las puntas micro-tijeras de punta roma con la mano derecha y cortar el bronquio y distal de la arteria pulmonar para el clip y retire pulmón izquierdo (Figura 3E). </li><li> Retire los retractores costales. </li><li> Utilice las pinzas contundentes curvas pellizcar hasta 1 cm de piel de calidad inferior a la incisien pero por encima del nivel del diafragma e insertar un angiocatéter 24 G a través de la piel y en la cavidad torácica izquierda (Figura 4A, B). </li><li> Utilice 5-0 prolene sutura colocar dos puntos separados alrededor de la 4 ª y 5 ª costillas para cerrar la cavidad torácica. </li><li> Retire los retractores de la piel. Utilice dos conjuntos de pinzas para aproximar la piel a lo largo de la longitud de la incisión y pegar la piel cerrada. </li><li> Conecte una jeringa luer-lock 3 ml al angiocatéter y eliminar el aire residual aplicando succión suave y retirar el angiocatéter. </li><li> Pegue la incisión en el cuello cerrado usando dos juegos de pinzas como antes. </li></ol> 5. Ratón Sham neumonectomía <ol><li> Exponer el pulmón izquierdo como se señala en el protocolo &quot;Ratón Neumonectomía&quot;. Levante la caja torácica con unas pinzas romas curvas para permitir que el aire en la cavidad torácica izquierda (Figura 5 A, B). </li><li> Coloque una G angiocatéter 24 en el tho izquierdacavidad Racic que arriba con cuidado de no dañar el pulmón izquierdo. </li><li> Utilizando 5-0 prolene sutura y teniendo cuidado de no perforar el pulmón (Figura 5C), colocar dos trozos de material de sutura en la 3ª / 4ª y 5ª / 6 espacios intermedios ª costilla (Figura 5D). Coloque las dos longitudes de material de sutura antes de atar a disminuir los riesgos de hernia pulmón izquierdo. Ate el material de sutura para hacer dos puntos separados (Figura 5E). </li><li> Pegue la piel sobre la incisión torácica, eliminar el aire residual con el angiocatéter y pegar la incisión en el cuello que el anterior. </li></ol> 6. Reanimación, Analgesia y Recuperación <ol><li> Apague el isoflurano, y administrar 0,1 mg / kg de buprenorfina y 0,5 ml de solución salina normal por vía subcutánea. </li><li> Cuando la respiración espontánea se reanuden, retire el tubo endotraqueal. </li><li> Observe ratón hasta que esté de nuevo ambulatorio. Caminar normalmente Resumit varios minutos después de la extracción del tubo endotraqueal. </li><li> Coloca el ratón en un 27 ° C incubadora (, 25% de oxígeno humidificado) para recuperar O / N. NOTA: Nos colocamos varias bolitas de comida humedecida con agua en el suelo de la jaula por primera 24 horas después de la cirugía. </li><li> Administrar 0,1 mg / kg de buprenorfina por inyección intraperitoneal dos veces al día durante tres días después de la cirugía. Tenga cuidado de no abrir el sitio quirúrgico al manipular animales. </li></ol> 7. Monitoreo Ratón <ol><li> Pesar ratones a 1, 3, 5, y 7 días después de la cirugía. </li></ol>

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The authors have nothing to disclose.

Hemos proporcionado la descripción más detallada de los procedimientos PNX PNX ratón y ratón del impostor reportados hasta la fecha. Hemos hecho el lector consciente de varios de los errores comunes que los investigadores están aprendiendo el procedimiento comúnmente se enfrentan, y que hemos descrito varias técnicas desarrolladas por nuestro laboratorio para mitigar estos riesgos. Otros laboratorios que utilizan este modelo pueden haber desarrollado otras modificaciones en la técnica o utilizar diferentes instr...

La función principal del pulmón es proporcionar para el oxígeno y el dióxido de carbono de intercambio entre un organismo y el ambiente. En los seres humanos, una serie de condiciones congénitas y adquiridas conducen a la reducción de la superficie pulmonar que se traduce en la función pulmonar. Aunque una serie de terapias tales como los corticosteroides inhalados, broncodilatadores, oxígeno suplementario, y la ventilación mecánica crónica se utilizan para mitigar las consecuencias de la función pulmonar 1-3, la terapia ideal para estas condiciones sería promover el recrecimiento del tejido pulmonar funcional - es decir, de pulmón regeneración. La regeneración de tejidos de mamíferos ha sido bien documentado. El Ratón espinoso africano puede regenerar grandes zonas de piel sin la formación de cicatrices 4. La falange distal en los seres humanos puede regenerarse después de una lesión o amputación 5-7. Siguiendo neumonectomía (PNX), el crecimiento pulmonar compensatorio se produce en ratones 8, 9 ratas, hacergs 10, y los seres humanos 11. Por definición, el crecimiento pulmonar compensatorio implica no sólo la expansión de los espacios aéreos existentes, pero volver a la tabicación de estos espacios aéreos ampliados con la expansión de la microcirculación 12 asociado. La expresión de genes ha demostrado que este modelo recapitula muchos de los eventos de señalización de desarrollo pulmonar 13. Cuatro semanas después de ratón PNX, área de la superficie alveolar es equivalente a la de animales operados simulados 14. En este manuscrito, se describe la PNX ratón y procedimientos simulados PNX.

<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.

. Una parcela de PNX y operación simulada pesos de los ratones se proporciona en la figura 6 En nuestras manos, la supervivencia es consistentemente 95-100% tanto para PNX y farsa neumonectomía. Para obtener una descripción de cómo el pulmón derecho re-crece en este modelo y la evolución en el tiempo previsto, nos remite al lector a los manuscritos de Gibney et al. 15 y Wang et al. 14 Varios errores comunes se deben evitar para r...

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.

Ratón neumonectomía Modelo de Compensatoria crecimiento pulmonar

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 vistas.  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.

Video: 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.

El desarrollo de bronquiolitis obliterante en un modelo murino de trasplante de pulmón ortotópico

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

Investigación

Medicina

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.

Un método estandarizado para medir el área de la superficie pulmonar interna mediante neumonectomía y prótesis de ratón

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

Biología del Desarrollo

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.

Medición de la curva de presión-volumen en mouse Pulmones

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.

Generación de un modelo de enfermedad pulmonar obstructiva crónica en ratones por la exposición repetida del ozono

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.

La neumonectomía izquierda combinado con Monocrotaline o Sugen como un modelo de hipertensión pulmonar en ratas

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.

Modelo experimental para evaluar la resolución de la neumonía

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

Inmunología e Infección

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.

Un modelo de trasplante de pulmón de rata de lesión por isquemia/reperfusión caliente: optimizaciones para mejorar los resultados

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.

Ventilación Normatérmica con Presión Negativa Perfusión Pulmonar Ex Situ : Evaluación de la Función Pulmonar y el Metabolismo

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>Una técnica sencilla de manguito para el trasplante de pulmón izquierdo murino

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.

Una técnica sencilla de manguito para el trasplante de pulmón izquierdo murino

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

Ratón neumonectomía Modelo de Compensatoria crecimiento pulmonar

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Un método estandarizado para medir el área de la superficie pulmonar interna mediante neumonectomía y prótesis de ratón

Medición de la curva de presión-volumen en mouse Pulmones

Generación de un modelo de enfermedad pulmonar obstructiva crónica en ratones por la exposición repetida del ozono

La neumonectomía izquierda combinado con Monocrotaline o Sugen como un modelo de hipertensión pulmonar en ratas

Modelo experimental para evaluar la resolución de la neumonía

Un modelo de trasplante de pulmón de rata de lesión por isquemia/reperfusión caliente: optimizaciones para mejorar los resultados

Ventilación Normatérmica con Presión Negativa Perfusión Pulmonar Ex Situ : Evaluación de la Función Pulmonar y el Metabolismo

Una técnica sencilla de manguito para el trasplante de pulmón izquierdo murino

Una técnica sencilla de manguito para el trasplante de pulmón izquierdo murino