Apoyado por el NIH HL-10342.

1. Preparación para el procedimiento Primero hay que obtener y preparar los siguientes elementos: <ol><li> La cánula. Para la intubación de 20-35 g ratones, se utiliza un 1 o 1,5 pulgadas de largo, calibre 20, catéter IV (BD Insylte, Sparks, MD o Optiva Jelco, Carlsbad, CA). Un catéter estéril nuevo se puede utilizar para cada ratón, pero catéteres también se pueden reutilizar después de la esterilización por inmersión en etanol al 70% durante la noche. Aunque ni la faringe ni tráquea del ratón es estéril procedimientos de limpieza adecuados, incluido el uso de guantes estériles e instrumentos, deben ser seguidas. </li><li> El cable de fibra óptica. Utilizamos ≈ 70 cm de cable de 0,5 mm Salida de Edmund Optics, pero la longitud no es crítica. Es importante asegurarse de que la fibra tiene su borde alisado, ya que después de cortar el cable a la longitud con una navaja de afeitar, el borde se deja relativamente agudo, y no se necesita mucho esfuerzo para perforar la pared traqueal. Sin embargo, is muy fácil para suavizar este borde sujetando la fibra de alrededor de 2 cm desde el extremo y luego hacer pequeños círculos durante unos segundos con los bordes de la punta tocando un trozo de papel de lija de grano 1.000 (véase la demostración en el vídeo y la Figura 1 de los Macdonald, et al. 10). El otro extremo se inserta a través de un tapón de goma. Esto se logra más fácilmente por primera empujando una aguja de calibre 18-a través del tapón, la inserción de la fibra óptica a través del orificio de aguja, a continuación, retirar la aguja. El tapón de goma está conectado a una fuente de luz halógena de 150 vatios (por ejemplo NCL-150, Volpi EE.UU., o cualquier otra fuente de luz o, incluso menos de 150 vatios). Es importante asegurarse de utilizar un tapón de goma de silicona (o otro material resistente al calor), ya que el corcho o caucho ordinario puede quemar cuando se encuentra tan cerca de la fuente de luz caliente. </li></ol> 2. Realización de la intubación <ol><li> Véanse las Figuras 1 y 2. Inserte cable de fibra óptica tediante un pequeño trozo de tubo de goma de silicona (≈ 0,8 mm x 4 mm OD, Cole-Palmer, EW-96410-13). Ate este tubo de goma bastante fuerte, al tiempo que permite el cable de fibra óptica para ajustar. Inserción del tubo de silicona cómodamente en el extremo de la cánula Luer fija la posición del cable de fibra óptica del interior de la cánula. Ajustar la posición del cable de fibra óptica para que se extienda a través de la cánula ≈ 4 mm por delante de la punta de la cánula. </li><li> Coloque el ratón anestesiado en un soporte vertical, suspendida por sus incisivos superiores (Figura 3). La mayoría de los investigadores a encontrar la mejor visualización con el lado ventral del ratón frente a sí mismos. Muy suavemente tire de la lengua y sostenga con el pulgar y el índice. El dedo medio se coloca entre el cuello y el soporte de plástico. La tracción sobre la lengüeta con el dedo índice y el pulgar se utiliza para abrir la boca, y para enderezar el camino intubación, el ángulo de la cabeza se ajusta con el dedo medio detrás del cuellose muestra en la Figura 3. </li><li> Con el cable de fibra óptica como fuente de luz y el introductor, empújelo a través de las cuerdas vocales visualizadas. Si los cables no están visibles, tire suavemente más duro en la lengua con el dedo medio como apoyo. Una vez introducido, avanzar la cánula ≈ 5 mm más allá. Luego, con mucho cuidado de no mover la cánula, retire el cable de fibra óptica. Lie el ratón hacia abajo y asegurar la cánula con un trozo de cinta y compatible con el cubo de la cánula en un trozo de plastilina (arcilla de modelado), como se muestra en la Figura 4. </li><li> El procedimiento del paso 3 no puede ser fácilmente enseñado o incluso demostrado, ya que es una operación en solitario. Sin embargo, por los ajustes sutiles de la tracción en la lengua y el apoyo detrás de la cabeza de casi todos los que tratan esto pronto encontrar la manera correcta de colocar el ratón para visualizar las cuerdas vocales. </li></ol>

<ol>
	<li>Brown, R. H., Walters, D. M., Greenberg, R. S., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+of+endotracheal+intubation+and+pulmonary+functional+assessment+for+repeated+studies+in+mice.">A method of endotracheal intubation and pulmonary functional assessment for repeated studies in mice.</a> J. Appl. Physiol. 87, 2362-2365 (1999).</li><li>Walters, D. M., Wills-Karp, M., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+cellular+profile+and+lung+function+with+repeated+bronchoalveolar+lavage+in+individual+mice.">Assessment of cellular profile and lung function with repeated bronchoalveolar lavage in individual mice.</a> Physiol. Genomics. 2, 29-36 (2000).</li><li>Rivera, B., Miller, S., Brown, E., Price, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+method+for+endotracheal+intubation+of+mice+and+rats+used+in+imaging+studies.">A novel method for endotracheal intubation of mice and rats used in imaging studies.</a> Contemporary Topics in Laboratory Animal Science / American Association for Laboratory Animal Science. 44, 52-55 (2005).</li><li>Hamacher, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microscopic+wire+guide-based+orotracheal+mouse+intubation:+description,+evaluation+and+comparison+with+transillumination.">Microscopic wire guide-based orotracheal mouse intubation: description, evaluation and comparison with transillumination.</a> Laboratory Animals. 42, 222-230 (2008).</li><li>Spoelstra, E. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+and+simple+method+for+endotracheal+intubation+of+mice.">A novel and simple method for endotracheal intubation of mice.</a> Laboratory Animals. 41, 128-135 (2007).</li><li>Zhao, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+technique+for+retrograde+intubation+in+mice.">A technique for retrograde intubation in mice.</a> Lab Animal. 35, 39-42 (2006).</li><li>Vergari, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+method+of+orotracheal+intubation+in+mice.">A new method of orotracheal intubation in mice.</a> European Review for Medical and Pharmacological Sciences. 8, 103-106 (2004).</li><li>Vergari, A., Polito, A., Musumeci, M., Palazzesi, S., Marano, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Video-assisted+orotracheal+intubation+in+mice.">Video-assisted orotracheal intubation in mice.</a> Laboratory Animals. 37, 204-206 (2003).</li><li>Hastings, R. H., Summers-Torres, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+Laryngoscopy+in+Mice.">Direct Laryngoscopy in Mice.</a> Contemporary Topics in Laboratory Animal Science / American Association for Laboratory Animal Science. 38, 33-35 (1999).</li><li>MacDonald, K. D., Chang, H. Y., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+simple+method+of+mouse+lung+intubation.">An improved simple method of mouse lung intubation.</a> J. Appl. Physiol. 106, 984-987 (2009).</li><li>Ewart, S. L., Levitt, R. C., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+system+mechanics+in+mice+measured+by+end-inflation+occlusion.">Respiratory system mechanics in mice measured by end-inflation occlusion.</a> Journal of Applied Physiology. 79, 560-566 (1995).</li><li>MacDonald, K. D., McKenzie, K. R., Mitzner, W., Zeitlin, P. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+Mechanics+in+Heterozygous+CF+Mice+after+Repeated+LPS+Dosing.">Lung Mechanics in Heterozygous CF Mice after Repeated LPS Dosing.</a> Am. J. Respir. Crit. Care Med. 175 (4), A930 (2007).</li></ol>

Ninguno de los autores tiene conflicto de intereses a revelar.

El procedimiento descrito aquí tiene varias ventajas. En primer lugar el aparato es sencillo y relativamente barato .. La fabricación del aparato no requiere ninguna herramienta especial o equipo costoso. El uso de un catéter que presenta también es la fuente de luz significa que uno nunca pierde de vista la apertura de la tráquea como el introductor se acerca a la apertura de la tráquea. El uso de un introductor de 0,5 mm también sirve para minimizar el trauma que podría ocurrir con una inserción inicial de un...

En 1999, Brown et al. Publicado un artículo que describe un método para la intubación de la pulmón de ratón 1. Dicha técnica tiene una utilidad considerable en hacer la función de repetición pulmonar o lavado broncoalveolar de ratones individuales en estudios longitudinales 2. Dado que el papel original, ha habido varios otros papeles que han descrito diferentes enfoques para la intubación ratón 3-9. Aunque todos estos métodos se pueden utilizar con éxito, por lo general requieren una formación considerable o coste. Uno de los principales problemas con la intubación es tal que cuando se acerca la cánula de intubación se acerca a la inserción tráquea pendiente, la propia cánula bloquea la luz y por lo tanto la visualización de donde tiene que ir. Por lo tanto, la inserción queda ciego en el momento más crítico. En este trabajo se muestra cómo eliminar sencilla y económica a este problema de visualización, lo que garantiza la intubación exitosa con relativamente poca formación oexperiencia.

<table border="1"><tbody><tr><td> Material </td><td> Empresa </td><td> Catálogo </td><td> Comentario </td></tr><tr><td> Intubación cánula </td><td> Insylte BD, Sparks, MD o Jelco Optiva, Carlsbad, CA </td><td></td><td> 1-in-largo, 20-gauge intravenosa (IV) de catéter </td></tr><tr><td> Cable de fibra óptica </td><td> Edmund Optics, Barrington, NJ </td><td> # NT02-542 </td><td> Alrededor de 2 pies de longitud de 0,5 mm de fibra óptica (fibra de Comunicación de plástico de grado). El borde del extremo de la fibra que se inserta en la tráquea debe ser suavemente redondeado mediante la celebración de la fibra ≈ 2 cm del extremo y luego hacer pequeños círculos mientras arrastra la punta durante unos segundos sobre papel de lija grano 1000 Emory. </td></tr><tr><td> Fuente de luz </td><td> Volpi </td><td> NCL-150 </td><td> Aunque utilizamos una fuente de halógeno de 150 W de luz, cualquier fuente de luz equivalente, aunque con mucho menor potenciapuede ser utilizado. </td></tr><tr><td> Tapón de goma de silicona </td><td></td><td></td><td> Un tapón de goma de silicona n º 1 se ajusta a la fuente de luz utilizada en esta demostración. Diferentes fuentes de luz puede requerir un tamaño diferente. </td></tr><tr><td> Silicona pequeño tubo de goma </td><td> Cole-Palmer </td><td> EW-96410-13 </td><td> A ≈ 1,5 cm trozo de tubo de goma de silicona (0,8 diámetro interior de 4 mm de diámetro exterior) </td></tr><tr><td> Caballete de soporte angular </td><td></td><td></td><td> La nuestra se constucted de plexiglás, pero cualquier material al que un hilo o alambre puede ser fijada a mantener el ratón en un ángulo casi vertical se puede utilizar. </td></tr></tbody></table>

a simple method of mouse lung intubation

Un procedimiento sencillo para intubar ratones para las mediciones de la función pulmonar tendría varias ventajas en estudios longitudinales con un número limitado de animales o caro. Una de las razones por las que esto no se hace de forma más rutinaria es que es relativamente difícil, a pesar de existir varios estudios publicados que describen la manera de lograrlo. En este trabajo se demuestra un procedimiento que elimina uno de los principales obstáculos asociados con este intubación, que de visualizar la tráquea durante todo el tiempo de intubación. El enfoque utiliza una fuente de 0,5 mm de luz de fibra óptica que sirve como un introductor para dirigir la cánula de intubación en la tráquea ratón. Se demuestra que es posible utilizar este procedimiento para medir la mecánica pulmonar en ratones individuales durante un transcurso de tiempo de al menos varias semanas. La técnica se puede configurar con relativamente poco gasto y experiencia, y que pueden ser rutinariamente a cabo con relativamente poca formación. Esto debería MAKE Es posible para cualquier laboratorio para llevar a cabo de forma rutinaria este intubación, lo que permite estudios longitudinales en ratones individuales, minimizando de este modo el número de ratones necesarios y aumentar el poder estadístico mediante el uso de cada ratón como su propio control.

Como una evaluación del método, hemos utilizado cuatro 20 semanas de edad BALB / c macho ratones con peso promedio (± SEM) de 27,7 ± 0,40 g. Se estudiaron en cinco semanas consecutivas, donde la resistencia pulmonar se midió utilizando un sistema como se ha descrito previamente 11. Cada ratón se anestesió con ketamina (100 mg / g BW) y xilazina (15 mg / g de peso corporal) en solución salina mediante inyección IP. Ellos fueron intubados entonces como se describe anteriormente. Si hay alguna duda sobr...

Watch this Scientific Journal Video about A Simple Method of Mouse Lung Intubation at JoVE.com

A Simple Method of Mouse Lung Intubation

Este artículo describe un método striaghforward y eficiente de intubación de ratones para las mediciones de la función pulmonar o la instilación pulmonar, que permite a los ratones para recuperar y se estudió en tiempos posteriores. El procedimiento consiste en una fuente de luz de fibra óptica de bajo costo que ilumina directamente la tráquea.

Un sencillo método de intubación ratón de pulmón

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Research

JoVE Journal

Medicine

50.6K vistas.  Johns Hopkins Bloomberg School of Public Health. Este artículo describe un método striaghforward y eficiente de intubación de ratones para las mediciones de la función pulmonar o la instilación pulmonar, que permite a los ratones para recuperar y se estudió en tiempos posteriores. El procedimiento consiste en una fuente de luz de fibra óptica de bajo costo que ilumina directamente la tráquea.

Video: A Simple Method of Mouse Lung Intubation

A Model of Disturbed Flow-Induced Atherosclerosis in Mouse Carotid Artery by Partial Ligation and a Simple Method of RNA Isolation from Carotid Endothelium

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified version of carotid partial ligation methods [1,2] to show that it acutely induces low and oscillatory flow conditions, two key characteristics of disturbed flow, in the mouse common carotid artery. Using this model, we have provided direct evidence that disturbed flow indeed leads to rapid and robust atherosclerosis development in Apolipoprotein E knockout mouse [3]. We also developed a method of endothelial RNA preparation with high purity from the mouse carotid intima [3]. Using this mouse model and method, we found that partial ligation causes endothelial dysfunction in a week, followed by robust and rapid atheroma formation in two weeks in a hyperlipidemic mouse model along with features of complex lesion formation such as intraplaque neovascularization by four weeks. This rapid in vivo model and the endothelial RNA preparation method could be used to determine molecular mechanisms underlying flow-dependent regulation of vascular biology and diseases. Also, it could be used to test various therapeutic interventions targeting endothelial dysfunction and atherosclerosis in considerably reduced study duration.

This describes a partial carotid ligation surgery, which causes disturbed flow conditions and subsequent atherosclerosis development (in two weeks) with intraplaque neo-vascularization (in four weeks) in the mouse common carotid artery. We also describe a novel method of RNA isolation from the carotid intima, providing high purity endothelial RNA.

Un modelo de Disturbed inducida por el flujo de Aterosclerosis en la arteria carótida del ratón por la ligadura parcial y un método sencillo para el aislamiento de ARN del endotelio carótida

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified ...

Investigación

Medicina

Guidelines for Elective Pediatric Fiberoptic Intubation

Fiberoptic intubation in pediatric patients is often required especially in difficult airways of syndromic patients i.e. Pierre Robin Syndrome. Small babies will desaturate very quickly if ventilation is interrupted mainly to high metabolic rate. We describe guidelines to perform a safe fiberoptic intubation while maintaining spontaneous breathing throughout the procedure. Steps requiring the use of propofol pump, fentanyl, glycopyrrolate, red rubber catheter, metal insuflation hook, afrin, lubricant and lidocaine spray are shown.

We describe guidelines to perform a safe and efficient elective fiberoptic intubation in pediatric patients while maintaining spontaneous ventilation.

Directrices para la intubación electiva de fibra óptica Pediátrica

Fiberoptic intubation in pediatric patients is often required especially in difficult airways of syndromic patients i.e. Pierre Robin Syndrome. Small ...

A Simple Guide Screw Method for Intracranial Xenograft Studies in Mice

The grafting of human tumor cells into the brain of immunosuppressed mice is an established method for the study of brain cancers including glioblastoma (glioma) and medulloblastoma. The widely used stereotactic approach only allows for the injection of a single animal at a time, is labor intensive and requires highly specialized equipment. The guide screw method, initially developed by Lal et al.,1 was developed to eliminate cumbersome stereotactic procedures. We now describe a modified guide screw approach that is rapid and exceptionally safe; both of which are critical ethical considerations. Notably, our procedure now incorporates an infusion pump that allows up to 10 animals to be simultaneously injected with tumor cells.
To demonstrate the utility of this procedure, we established human U87MG glioma cells as intracranial xenografts in mice, which were then treated with AMG102; a fully human antibody directed to HGF/scatter factor currently undergoing clinical evaluation2-5. Systemic injection of AMG102 significantly prolonged the survival of all mice with intracranial U87MG xenografts and resulted in a number of complete cures. 
This study demonstrates that the guide screw method is an inexpensive, highly reproducible approach for establishing intracranial xenografts. Furthermore, it provides a relevant physiological model for validating novel therapeutic strategies for the treatment of brain cancers.

In order to evaluate novel therapeutic paradigms for the treatment of glioma, physiological relevant models are essential. We utilize an implantable guide screw procedure for establishment of intracranial xenograft models that is more rapid and safer than stereotactic approaches.

Una guía sencilla Método de tornillo para intracraneal Estudios xenoinjerto en ratones

The grafting of human tumor cells into the brain of immunosuppressed mice is an established method for the study of brain cancers including glioblastoma ...

Noninvasive Intratracheal Intubation to Study the Pathology and Physiology of Mouse Lung

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular disease and for developing therapeutic intervention. With the increasing availability of knockout and transgenic derivatives, together with a vast amount of genetic information, mice provide one of the best models to study the molecular mechanisms underlying the pathology and physiology of lung diseases. Inhalation, intranasal instillation, intratracheal instillation, and intratracheal intubation are the most widely used techniques by a number of investigators to administer materials of interest to mouse lungs. There are pros and cons for each technique depending on the goals of a study. Here a noninvasive intratracheal intubation method that can directly deliver exogenous materials to mouse lungs is presented. This technique was applied to administer bleomycin to mouse lungs as a model to study pulmonary fibrosis.

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular disease and for developing therapeutic intervention. Here a noninvasive intratracheal intubation method that can directly deliver exogenous materials to mouse lungs is presented.

No Invasiva Intratraqueal Intubación para Estudiar la Patología y Fisiología de pulmón de ratón

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular ...

Implantation of Fibrin Gel on Mouse Lung to Study Lung-specific Angiogenesis

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and repair damage in simple tissues in the orthopedic and periodontal fields. However, attempts to regenerate the structures and functions of more complex three-dimensional (3D) organs such as lungs have not been very successful because the biological processes of organ regeneration have not been well explored. It is becoming clear that angiogenesis, the formation of new blood vessels, plays key roles in organ regeneration. Newly formed vasculatures not only deliver oxygen, nutrients and various cell components that are required for organ regeneration but also provide instructive signals to the regenerating local tissues. Therefore, to successfully regenerate lungs in an adult, it is necessary to recapitulate the lung-specific microenvironments in which angiogenesis drives regeneration of local lung tissues. Although conventional in vivo angiogenesis assays, such as subcutaneous implantation of extracellular matrix (ECM)-rich hydrogels (e.g., fibrin or collagen gels or Matrigel - ECM protein mixture secreted by Engelbreth-Holm-Swarm mouse sarcoma cells), are extensively utilized to explore the general mechanisms of angiogenesis, lung-specific angiogenesis has not been well characterized because methods for orthotopic implantation of biomaterials in the lung have not been well established. The goal of this protocol is to introduce a unique method to implant fibrin gel on the lung surface of living adult mouse, allowing for the successful recapitulation of host lung-derived angiogenesis inside the gel. This approach enables researchers to explore the mechanisms by which the lung-specific microenvironment controls angiogenesis and alveolar regeneration in both normal and pathological conditions. Since implanted biomaterials release and supply physical and chemical signals to adjacent lung tissues, implantation of these biomaterials on diseased lung can potentially normalize the adjacent diseased tissues, enabling researchers to develop new therapeutic approaches for various types of lung diseases.

Recapitulation of the organ-specific microenvironment, which stimulates local angiogenesis, is indispensable for successful regeneration of damaged tissues. This report demonstrates a novel method to implant fibrin gels on the lung surface of living mouse in order to explore how the lung-specific microenvironment modulates angiogenesis and alveolar regeneration in adult mouse.

El implante de gel de fibrina en pulmón de ratón para el Estudio de la angiogénesis pulmón específico

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and ...

Intubation-mediated Intratracheal (IMIT) Instillation: A Noninvasive, Lung-specific Delivery System

Respiratory disease studies typically involve the use of murine models as surrogate systems. However, there are significant physiologic differences between the murine and human respiratory systems, especially in their upper respiratory tracts (URT). In some models, these differences in the murine nasal cavity can have a significant impact on disease progression and presentation in the lower respiratory tract (LRT) when using intranasal instillation techniques, potentially limiting the usefulness of the mouse model to study these diseases. For these reasons, it would be advantageous to develop a technique to instill bacteria directly into the mouse lungs in order to study LRT disease in the absence of involvement of the URT. We have termed this lung specific delivery technique intubation-mediated intratracheal (IMIT) instillation. This noninvasive technique minimizes the potential for instillation into the bloodstream, which can occur during more invasive traditional surgical intratracheal infection approaches, and limits the possibility of incidental digestive tract delivery. IMIT is a two-step process in which mice are first intubated, with an intermediate step to ensure correct catheter placement into the trachea, followed by insertion of a blunt needle into the catheter to mediate direct delivery of bacteria into the lung. This approach facilitates a &#62;98% efficacy of delivery into the lungs with excellent distribution of reagent throughout the lung. Thus, IMIT represents a novel approach to study LRT disease and therapeutic delivery directly into the lung, improving upon the ability to use mice as surrogates to study human respiratory disease. Furthermore, the accuracy and reproducibility of this delivery system also makes it amenable to Good Laboratory Practice Standards (GLPS), as well as delivery of a wide range of reagents which require high efficiency delivery to the lung.

Intubation-mediated Intratracheal IMIT Instillation: A Noninvasive, Lung-specific Delivery System

Intubation-mediated intratracheal (IMIT) instillation of reagents is an excellent, noninvasive method for studying respiratory disease, as well as a method for instilling therapeutic reagents directly into the lung. It is a rapid and highly reproducible method which is suitable for preclinical testing.

La intubación mediada intratraqueal (IMIT) instilación: A, Sistema de Entrega de pulmón-específica no invasiva

Respiratory disease studies typically involve the use of murine models as surrogate systems. However, there are significant physiologic differences ...

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.

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 ...

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. As this technology expands and improves, the ability to potentially evaluate and improve the quality of substandard lungs prior to transplant is a critical need. In order to more rigorously evaluate these approaches, a reproducible animal model needs to be established that would allow for testing of improved techniques and management of the donated lungs as well as to the lung-transplant recipient. In addition, an EVLP animal model of associated pathologies, e.g., ventilation induced lung injury (VILI), would provide a novel method to evaluate treatments for these pathologies. Here, we describe the development of a rat EVLP lung program and refinements to this method that allow for a reproducible model for future expansion. We also describe the application of this EVLP system to model VILI in rat lungs. The goal is to provide the research community with key information and &#8220;pearls of wisdom&#8221;/techniques that arose from trial and error and are critical to establishing an EVLP system that is robust and reproducible.

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. Here, we describe the development of a rat EVLP program and refinements that allow for a reproducible model for future expansion.

Método de aislados<em&gt; Ex Vivo</em&gt; Perfusión pulmonar en un modelo de rata: Lecciones Aprendidas de Desarrollo de un Programa EVLP Rata

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed ...

A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe the use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations of superior and uniform quality to that which can be achieved by hand. The device comprises a base that holds 4 stainless steel rods and a top, which acts a cutting guide. The rods are inserted into the lumen of the small intestine [divided into thirds] and the colon. The rods and samples are then placed over a piece of filter paper or card into the holding slots in the base of the device. The top of the device is then positioned and serves as a cutting guide. The two angled sections in the center of the top piece are used to guide a knife or scalpel and cut the intestines longitudinally on the top of the rods. Once the intestinal sections have been cut, the top is removed and the card,&#160;tissue and rods gently removed from the device and placed on the bench. The rods are then gently rolled sideways to flatten and stick the intestinal segments onto the underlying piece of filter paper or card. The final preparation can then be examined or fixed and stored for later analysis. The preparations are invaluable for the study of intestinal changes in normal or genetically modified mouse models. The preparations have been used for the study and quantification of the effects of inflammation (colitis), damage, pre-cancerous lesions (aberrant crypt foci (ACFs) and mucin depleted foci (MDFs)) and polyps or tumors.

The use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations is described.

Un dispositivo sencillo de preparar rápidamente enteros Montes del Intestino Ratón

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe ...

An Improved Method for Rapid Intubation of the Trachea in Mice

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity is a commonly used model to study both acute lung injury and fibrosis, and multiple methods have been developed for administering bleomycin (and other toxic agents) into the lungs. However, many of these approaches, such as transtracheal instillation, have inherent drawbacks, including the need for strong anesthetics and survival surgery. This paper reports a quick, reproducible method of intratracheal intubation that involves mild inhaled anesthesia, visualization of the trachea, and the use of a surrogate spirometer to confirm exposure. As a proof of concept, 8-12 week old C57BL/6 mice were administered either 2.0 U/kg of bleomycin or an equivalent volume of PBS, and both damage and fibrotic endpoints were measured post-exposure. This procedure allows researchers to treat a large cohort of mice in a relatively short period with little expense and minimal post-procedure care.

This article presents a rapid and simple method for administering bleomycin directly into the mouse trachea via intubation. Key advantages of this method are that it is highly reproducible, easy to master, and does not require specialized equipment or lengthy recovery times.

Un método mejorado para la rápida intubación de la tráquea en ratones

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...