Unterstützt durch NIH HL-10342.

Ein. Vorbereitung für die Vorgehensweise Man muss zuerst erhalten und bereiten die folgenden Elemente: <ol><li> Die Kanüle. Für Intubation von 20-35 g Mäusen, verwenden wir eine 1 oder 1,5 Zoll lang, 20 Gauge IV-Katheter (BD Insylte, Sparks, MD oder Jelco Optiva, Carlsbad, CA). Ein neuer steriler Katheter kann für jede Maus verwendet werden, aber Kathetern können auch nach der Sterilisation durch Einweichen in 70% Ethanol über Nacht wiederverwendet werden. Obwohl weder der Rachen noch Trachea der Maus ist steril richtigen Sauberkeit Verfahren, einschließlich der Verwendung von sterilen Handschuhen und Instrumente angewendet werden sollten. </li><li> Die faseroptische Kabel. Wir verwenden ≈ 70 cm von 0,5 mm optische Kabel von Edmund Optics, aber die Länge ist nicht kritisch. Es ist wichtig, sicherzustellen, dass die Faser ihrem Rand geglättet, da nach dem Schneiden des Kabels zu Länge mit einer Rasierklinge, die Kante nach links relativ scharfe, und es nimmt nicht viel Aufwand zu durchstechen die Trachealwand. Allerdings ist es is sehr leicht, diese Kante, indem die Faser etwa 2 cm vom Ende und dann machen kleine Kreise für ein paar Sekunden mit den Kanten der Spitze berühren ein Stück 1.000 Schmirgelpapier (siehe Demonstration in der Video-und der Abbildung 1 zu glätten MacDonald, et al. 10). Das andere Ende ist durch einen Gummistopfen gesteckt. Dies geschieht am einfachsten, indem erste Schiebeglied eine 18-Gauge-Nadel durch den Stopfen, Einführen des Lichtwellenleiters durch die Nadelbohrung, dann Zurückziehen der Nadel erfolgt. Der Gummistopfen mit einer 150 Watt Halogen-Lichtquelle angeschlossen ist (zB NCL-150, Volpi USA, oder jede andere Lichtquelle oder sogar weniger als 150 Watt). Es ist darauf zu achten, um einen Stopfen aus Silikongummi (oder anderen hitzebeständigen Material) zu verwenden, da gewöhnlichen Gummi oder Kork kann verbrennen, wenn sich so nahe an der heißen Lichtquelle. </li></ol> 2. Durchführung der Intubation <ol><li> Siehe Abbildungen 1 und 2. Legen Glasfaserkabel turch ein kurzes Stück aus Silikon-Kautschuk-Schlauch (≈ 0,8 mm ID x 4 mm OD, Cole-Palmer, EW-96410-13). Tie dieses Gummischlauch ziemlich eng, solange noch die Glasfaserkabel angepasst werden. Einsetzen der Silikonschlauch gut in der Luer Ende der Kanüle fixiert das Glasfaserkabel Position in der Kanüle. Passen Sie die Position des Fiberkabels so dass es sich durch die Kanüle ≈ 4 mm vor der Kanülenspitze. </li><li> Platzieren Sie die anästhesierten Maus auf einem vertikalen Träger, der mit seinem oberen Schneidezähne (Abbildung 3) suspendiert. Die meisten Ermittler finden Sie die besten Visualisierung mit der Bauchseite der Maus vor sich. Sehr vorsichtig die Zunge heraus und halten Sie mit Daumen und Zeigefinger. Der Mittelfinger ist zwischen dem Hals und Kunststoffträger platziert. Traction auf der Zunge mit dem Daumen und Zeigefinger wird verwendet, um den Mund zu öffnen, und die Intubation Weg begradigen, wird der Winkel des Kopfes mit dem Mittelfinger hinter dem Hals eingestelltin 3 gezeigt. </li><li> Mit dem LWL-Kabel als Lichtquelle und Einführhilfe, schieben Sie es durch die visualisierte Stimmbänder. Wenn die Kabel nicht sichtbar sind, sanft schwerer ziehen auf der Zunge mit dem Mittelfinger als Unterstützung. Nach dem Einlegen voran die Kanüle ≈ 5 mm weiter. Dann sehr vorsichtig nicht zu bewegen die Kanüle zurückziehen Glasfaserkabel. Liegen die Maus nach unten und sichern Sie die Kanüle mit einem Stück Klebeband und unterstützen die Kanülenansatz auf ein Stück Knetmasse (Modelliermasse), wie in Abbildung 4 dargestellt. </li><li> Das Verfahren in Schritt 3 nicht leicht gelehrt oder sogar nachgewiesen werden, da es ein Solobetrieb ist. Doch durch subtile Anpassungen der Traktion auf der Zunge und der Unterstützung hinter dem Kopf fast alle, die versuchen, diese bald den richtigen Weg finden, um die Maus zu positionieren, um die Stimmbänder zu visualisieren. </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>

Keiner der Autoren keine Interessenkonflikte offenlegen.

Das hier beschriebene Verfahren hat mehrere Vorteile. Zunächst wird die Vorrichtung ist einfach und relativ kostengünstig .. Die Herstellung des Gerätes erfordert keine speziellen Werkzeuge oder teure Ausrüstung. Die Verwendung eines Katheters führt, daß auch die Lichtquelle bedeutet, daß man nie verliert der Trachealöffnung als die Einführeinrichtung die Trachealöffnung nähert. Die Verwendung eines 0,5 mm Einführungsvorrichtung dient auch dazu, Trauma, das mit einem anfänglichen Einsetzen eines größeren ...

In 1999 veröffentlichte Brown et al. Ein Papier beschreibt eine Methode zur Intubation der Mauslunge 1. Eine solche Technik hat erhebliche Utility dabei wiederholen Lungenfunktion oder Bronchalveolarlavages in einzelnen Mäusen in Längsschnittstudien 2. Seit diesem ursprünglichen Papier, gab es mehrere andere Papiere, die verschiedene Ansätze beschrieben haben, um mit der Maus Intubation 3-9. Während all dieser Verfahren erfolgreich verwendet werden kann, sie in der Regel nur mit erheblichem Ausbildung oder einsehbar. Eines der Hauptprobleme bei solchen Intubation ist, dass die Intubationskanüle Ansätze nähert sich die Luftröhre anstehenden Insertion, sperrt die Kanüle selbst das Licht und damit die Visualisierung von wo es gehen muss. Somit wird das Einführen blinden am meisten kritischen Zeitpunkt. In diesem Papier zeigen wir, wie einfach und kostengünstig zu beseitigen diese Visualisierung Problem, wodurch eine erfolgreiche Intubation mit relativ wenig Training oderErfahrung.

<table border="1"><tbody><tr><td> Material </td><td> Firma </td><td> Katalog # </td><td> Kommentar </td></tr><tr><td> Intubationskanüle </td><td> BD Insylte, Sparks, MD oder Jelco Optiva, Carlsbad, CA </td><td></td><td> 1-in.-länge, 20-Gauge intravenöse (IV)-Katheter </td></tr><tr><td> LWL- </td><td> Edmund Optics, Barrington, NJ </td><td> # NT02-542 </td><td> Etwa 2-ft Länge von 0,5-mm-Glasfaser (Communication hochwertigem Kunststoff-Faser). Der Rand des Faserendes, die in die Luftröhre eingeführt wird, sollte vorsichtig, indem die Faser ≈ 2 cm vom Ende und dann machen kleine Kreise, während Sie den Tipp für ein paar Sekunden auf 1.000-grit emory Papier abgerundet werden. </td></tr><tr><td> Light Source </td><td> Volpi </td><td> NCL-150 </td><td> Obwohl wir einen 150-W-Halogen-Lichtquelle, einer äquivalenten Lichtquelle, selbst mit sehr viel geringeren Wattzahlkönnen verwendet werden. </td></tr><tr><td> Silikon Gummistopfen </td><td></td><td></td><td> A # 1 Silikonkautschuk Stopfen passt der Lichtquelle in dieser Demonstration verwendet. Verschiedene Lichtquellen können eine unterschiedliche Größe. </td></tr><tr><td> Kleine Silikonkautschuk Rohr </td><td> Cole-Palmer </td><td> EW-96.410-13 </td><td> A ≈ 1,5 cm großes Stück aus Silikonkautschuk Rohrleitung (Innendurchmesser 0,8, 4 mm Außendurchmesser) </td></tr><tr><td> Angled Ständer </td><td></td><td></td><td> Ours ist aus Plexiglas constucted, sondern jedes Material, an dem ein Faden oder Draht kann angebracht, um die Maus an einer nahezu senkrechten Winkel zu halten werden können verwendet werden. </td></tr></tbody></table>

a simple method of mouse lung intubation

Ein einfaches Verfahren, um Mäuse für Lungenfunktion Messungen intubieren hätte mehrere Vorteile in Längsschnittstudien mit einer begrenzten oder teure Tier. Einer der Gründe, dass dies nicht geschieht routinemäßig mehr ist, dass es relativ schwierig ist, obwohl es mehrere veröffentlichte Studien, die Wege dies zu erreichen beschreiben. In diesem Papier zeigen wir ein Verfahren, das eine der größten Hürden mit dieser Intubation assoziiert, dass der Visualisierung die Luftröhre während der gesamten Zeit der Intubation beseitigt. Der Ansatz nutzt eine 0,5 mm LWL-Lichtquelle, die als Einführer die Intubation Kanüle in die Luftröhre Maus lenken dient. Wir zeigen, dass es möglich ist, dieses Verfahren zu verwenden, um Lungenmechanik in einzelnen Mäusen über einen Zeitverlauf von mindestens mehreren Wochen zu messen. Die Technik kann mit relativ geringem Aufwand und Erfahrung eingestellt werden, und es kann routinemäßig mit relativ wenig Ausbildung erreicht werden. Dies sollte makê es jedem Labor routinemäßig Durchführung dieses Intubation, wodurch Längsschnittstudien in einzelnen Mäusen, wodurch eine Minimierung der Anzahl von Mäusen benötigt und Erhöhen der statistische unter Verwendung jeder Maus als seine eigene Kontrolle.

Als Bewertung des Verfahrens verwendeten wir vier 20 Wochen alte männliche BALB / c-Mäuse mit einem durchschnittlichen Gewicht (± SEM) von 27,7 ± 0,40 g auf. Sie wurden auf fünf aufeinanderfolgende Wochen untersucht, wobei die Lungenwiderstand wurde unter Verwendung eines Systems, wie zuvor beschrieben 11. Jede Maus wurde mit Ketamin (100 ug / g BW) und Xylazin (15 ug / g BW) in Kochsalzlösung über IP-Injektion. Sie wurden dann intubiert wie oben beschrieben. Wenn es irgendeinen Zweifel, ob die Kanüle...

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

A Simple Method of Mouse Lung Intubation

Dieses Papier beschreibt eine striaghforward und effiziente Methode zur Intubation Mäusen Lungenfunktion Messungen oder pulmonale Instillation, dass die Mäuse sich zu erholen und zu späteren Zeitpunkten untersucht werden können. Das Verfahren beinhaltet eine kostengünstige LWL-Lichtquelle, die direkt beleuchtet die Luftröhre.

Eine einfache Methode der Maus Lung Intubation

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

50.5K Aufrufe.  Johns Hopkins Bloomberg School of Public Health. Dieses Papier beschreibt eine striaghforward und effiziente Methode zur Intubation Mäusen Lungenfunktion Messungen oder pulmonale Instillation, dass die Mäuse sich zu erholen und zu späteren Zeitpunkten untersucht werden können. Das Verfahren beinhaltet eine kostengünstige LWL-Lichtquelle, die direkt beleuchtet die Luftröhre.

Serie: 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.

Ein Modell des Disturbed Flow-Atherosklerose in Maus Halsschlagader durch partielle Ligation und eine einfache Methode zur RNA-Isolierung aus Carotid Endothelium

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

Forschung

Medizin

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.

Richtlinien für die Wahlfächer Pediatric Fiberoptische Intubation

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.

Eine einfache Anleitung Screw Verfahren zur Intrakranielle Xenograft Studien an Mäusen

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.

Nichtinvasive Intratracheale Intubation, die Pathologie und Physiologie der Maus Lung Study

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.

Die Implantation von Fibrin Gel auf Maus Lung Lung spezifischen Angiogenese Studieren

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.

Intubation vermittelte Intratracheale (IMIT) Instillation: eine nichtinvasive, lungenspezifischen Delivery System

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.

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 ...

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.

Methode Isolated<em&gt; Ex vivo</em&gt; Lung Perfusion in einem Rattenmodell: Lehren aus Entwicklung einer Ratte EVLP Programm gelernt

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.

Eine einfache Vorrichtung, um schnell vorbereiten Whole Mounts des Mausdarm

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.

Ein verbessertes Verfahren zur schnellen Intubation der Luftröhre in Mice

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