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.

utilisation déclaration animale:: NOTE Toutes les procédures dans cette étude ont été menées avec l&#39;approbation et en suivant les lignes directrices du Comité des soins (IACUC) Institutionnel utilisation des animaux et à l&#39;Hôpital pour enfants de Cincinnati. Huit semaines C57BL / 6J mâles ont été obtenus auprès de Jackson Laboratories (Bar Harbor, ME) et on les laisse se acclimater pendant une semaine avant utilisation. Jusqu&#39;à la chirurgie, les animaux ont été logés dans un établissement de barrière exempts d&#39;agents pathogènes et à condition chow autoclave et filtré de l&#39;eau ad libdium. Chaque cage de la souris a été fournie avec un air dédié et de l&#39;eau, et les chambres ont été maintenus sur un cycle jour-nuit de 12 heures. Après récupération de la chirurgie, les souris ont été maintenues dans des cages avec des sommets filtrées, à condition autoclave chow ad libitum, et à condition que l&#39;eau filtrée à partir d&#39;une bouteille d&#39;eau. 1. Préparation des Instruments <ol><li> Faire six rétracteurs de peau en utilisant trombones et des broches. Twist redressa trombones sur les tiges des broches de papier, ll&#39;avant-toit d&#39;un fil d&#39;acier droite 5 cm à une extrémité et faire un seul 0,5 cm &quot;U&quot; en forme de crochet à l&#39;extrémité du fil. </li><li> Faire quelques 15 x 15 cm draps chirurgicaux carrés en utilisant une pellicule de plastique. Préparer une vinaigrette par souris. Mettez un tour de papier entre chaque wrap. </li><li> Stériliser tous les instruments chirurgicaux avec une pile de tuiles 12 x 12 pouces de liège, de la gaze et tampons de coton-tige. </li></ol> 2. Préparation de la souris <ol><li> Induire une anesthésie avec 2% d&#39;isoflurane. Peser animal. </li><li> Dans une zone de préparation chirurgicale dédiée raser thorax gauche et le cou avec le rasoir électrique. </li><li> Appliquez une goutte de la déchirure pommade artificielle aux yeux de la souris. </li><li> Décontaminer cou et du thorax gauche avec de la chlorhexidine et d&#39;alcool isopropylique. Répéter deux fois plus. </li></ol> 3. Souris Oro-intubation trachéale et la ventilation mécanique <ol><li> Avoir une place de technicien chirurgicale non stérile décubitus dorsal de la souris dans la zone chirurgicale préchauffé.</li><li> Confirmez profondeur de l&#39;anesthésie en documentant l&#39;absence d&#39;une réponse à la patte pincée. </li><li> Après le lavage des mains et enfiler vêtements chirurgicaux, masque et chapeau, enfiler des gants chirurgicaux stériles. </li><li> Après le drapage et utilisant une technique aseptique, faire une incision verticale de 1 cm sur la mi-partie antérieure du cou pour exposer le larynx. Rétracter légèrement les muscles de la sangle avec courbes dentelée, 10 cm pince et exposer le larynx et la trachée en écartant les muscles de la sangle avec la pointe d&#39;une paire de ciseaux droites. </li><li> Insérez oralement un 22 G émoussée pointe angiocathéter dans la mi-trachée (figure 1A) et de confirmer visuellement placement (figure 1B). Maintenir l&#39;anesthésie et ventiler en utilisant 1-3% d&#39;isoflurane travers rongeurs ventilateur (225 pi par course; 200 Stokes par min). Employer une limite de pression de 15 cm H 2 O. </li></ol> 4. Souris Pneumonectomie <ol><li> Placez la souris dans la position latérale droite de décubitus avec le dos de la souris facel&#39;opérateur (à gauche vers le haut). Utilisez un emballage de plastique auto-obturant comme un champ stérile. Coupe à travers le drap, utiliser contondants ciseaux courbes pressenti pour faire un 2 cm de long coupe parallèle aux côtes lors de la 4 ème et 5 ème espace intercostal. Insérez l&#39;extrémité incurvée ciseaux émoussés et disséquer la peau loin des côtes sous-jacents et muscles intercostaux. </li><li> Rentrer la peau avec quatre écarteurs pour exposer un carré 1,5 x 1,5 cm de la fenêtre chirurgicale (figure 2A). Fixez les écarteurs au conseil de liège. </li><li> Disséquer vers les côtes en utilisant des pinces courbes, et d&#39;utiliser une pointe de la pince courbes d&#39;entrer dans la cavité thoracique. </li><li> Avec la pointe émoussée micro-ciseaux, utiliser la lame inférieure pour entrer dans la cavité thoracique. Faire une incision de 0,5 cm entre les côtes et la répétition dans la direction opposée. </li><li> En utilisant les deux écarteurs restants, ouvrir le thorax dans l&#39;axe antéro-postérieur et sécurisé les rétracteurs à le panneau de liège (figure 2B). </li> &lt;li&gt; En utilisant des pinces atraumatiques de courbes dans la main gauche, saisir le poumon gauche et de déplacer la partie supérieure du poumon gauche et en bas latéralement à travers la thoracotomie jusqu&#39;à l&#39;artère pulmonaire gauche et des bronches sont exposés (figure 3A, B). </li><li> Tenir le titane vasculaire microclip applicateur chargé dans la main droite avec le corps de l&#39;applicateur dans la paume et pointe recourbée pointant loin de la paume (figure 3C), faites glisser le bout de l&#39;applicateur dans le thorax le long de la courbure de la face postérieure de la gauche poumon et clip la bronche gauche et l&#39;artère pulmonaire (Figure 3D). </li><li> Retirez l&#39;applicateur mais garder le poumon gauche rétractée. Saisir la pointe micro-ciseaux émoussés avec la main droite et couper la bronche et distale de l&#39;artère pulmonaire au clip et retirer poumon gauche (figure 3E). </li><li> Retirez les rétracteurs de côtes. </li><li> Utilisez la pince contondants courbes pour pincer la peau de 1 cm inférieurs au incisisur mais au-dessus du niveau de la membrane et insérer un angiocathéter G 24 à travers la peau et dans la cavité thoracique gauche (figure 4A, B). </li><li> Utilisez 5-0 prolène suture de placer deux points séparés dans le 4 ème et 5 ème nervures pour fermer la cavité thoracique. </li><li> Retirez les rétracteurs de la peau. Utiliser deux ensembles de pince pour rapprocher la peau le long de la longueur de l&#39;incision et de la colle la peau fermée. </li><li> Connectez un 3 ml luer-lock seringue à la angiocathéter et retirer l&#39;air résiduel en appliquant une aspiration douce et le retrait de la angiocathéter. </li><li> Coller l&#39;incision du cou fermée à l&#39;aide de deux ensembles de pinces comme avant. </li></ol> 5. Souris Sham Pneumonectomie <ol><li> Exposer le poumon gauche comme indiqué dans le protocole &quot;Souris Pneumonectomie&quot;. Soulevez la cage thoracique avec le forceps émoussé courbes pour permettre à l&#39;air dans la cavité de la poitrine gauche (figure 5A, B). </li><li> Placez un G angiocathéter 24 dans le tho gauchecavité Racic comme ci-dessus en faisant attention de ne pas blesser le poumon gauche. </li><li> Utilisation 5-0 prolène suture et en faisant attention à ne pas percer le poumon (Figure 5C), placez deux longueurs de matériau de suture dans le 3 e / 4 e et 5 e / 6 e interstices de nervure (Figure 5D). Placer les deux longueurs de matériau de suture avant de l&#39;attacher à réduire les risques de hernie du poumon gauche. Attachez le matériel de suture pour faire deux points de suture interrompue (figure 5E). </li><li> Collez la peau sur l&#39;incision thoracique, retirer l&#39;air résiduel avec le angiocathéter, et collez l&#39;incision du cou comme ci-dessus. </li></ol> 6. réanimation, analgésie, et récupération <ol><li> Éteignez l&#39;isoflurane, et administrer 0,1 mg / kg de buprénorphine et 0,5 ml de solution saline normale voie sous-cutanée. </li><li> Lorsque la respiration spontanée reprendre, retirer la sonde d&#39;intubation. </li><li> Observez la souris jusqu&#39;à ce qu&#39;il soit à nouveau ambulatoire. Marcher RESUM généralementes plusieurs minutes après le retrait de la sonde endotrachéale. </li><li> Placez la souris dans un 27 ° C incubateur (humidifié, 25% d&#39;oxygène) pour récupérer O / N. NOTE: Nous mettons plusieurs pastilles de chow mouillée avec de l&#39;eau sur le sol de la cage pour les 24 premières heures après la chirurgie. </li><li> Administrer 0,1 mg / kg de buprénorphine par injection intraperitoneale deux fois par jour pendant trois jours après la chirurgie. Prenez soin de ne pas ouvrir le site chirurgical lors de la manipulation des animaux. </li></ol> 7. Suivi de la souris <ol><li> Peser les souris à 1, 3, 5, et 7 jours après la chirurgie. </li></ol>

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The authors have nothing to disclose.

Nous avons fourni la description la plus détaillée des procédures de PNX de PNX de la souris et de la souris faux rapportés à ce jour. Nous avons fait le lecteur au courant de plusieurs des écueils les plus courants que les enquêteurs de la procédure d&#39;apprentissage rencontrent couramment, et nous avons présenté plusieurs techniques développées par notre laboratoire pour atténuer ces écueils. D&#39;autres laboratoires qui utilisent ce modèle peuvent avoir développé d&#39;autres modifications de tech...

La fonction principale du poumon est de fournir de l&#39;oxygène et du dioxyde de carbone échange entre un organisme et l&#39;atmosphère. Chez l&#39;homme, une multitude de conditions congénitales et acquises entraîner une diminution de la surface des poumons qui résulte de la fonction pulmonaire réduite. Bien que de nombreuses thérapies tels que les corticostéroïdes inhalés, les bronchodilatateurs, l&#39;oxygène supplémentaire, et la ventilation mécanique chronique sont utilisés pour atténuer les conséquences de la fonction pulmonaire 1-3, la thérapie idéale pour ces conditions serait favoriser la repousse du tissu pulmonaire fonctionnelle - ce est à dire, du poumon régénération. La régénération de tissu de mammifère a été bien documentée. La tortue-souris africaine peut se régénérer de vastes zones de la peau sans la formation de cicatrices 4. La phalange distale chez l&#39;homme peut se régénérer après une lésion ou une amputation 5-7. Suite pneumonectomies (PNX), la croissance du poumon compensatoire se produit chez des souris 8, 9 rats, doGS 10, 11 et les humains. Par définition, la croissance du poumon compensatoire implique non seulement l&#39;expansion des espaces aériens existants, mais re-cloisonnement de ces espaces aériens élargis avec l&#39;expansion de la microcirculation associée 12. analyse de l&#39;expression génique a montré que ce modèle récapitule plusieurs des événements de signalisation du développement pulmonaire 13. Quatre semaines après PNX souris, la surface alvéolaire est équivalente à celle des animaux opérés factices 14. Dans ce manuscrit, nous décrivons la PNX de la souris et les procédures PNX faux.

<table border="0" cellpadding="0" cellspacing="0" width="1052">
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:263px;">Name of the reagent</td>
			<td style="width:263px;">Company</td>
			<td style="width:263px;">Catalogue number</td>
			<td style="width:263px;">Comments (optional)</td>
		</tr>
		<tr height="101">
			<td height="101" style="height:101px;width:263px;">6-inch Vascular Clip Applicator</td>
			<td style="width:263px;">Teleflex Medical (WECK)</td>
			<td style="width:263px;">137062</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="101">
			<td height="101" style="height:101px;width:263px;">Horizon Small Titanium Red Clip</td>
			<td style="width:263px;">Teleflex Medical (WECK)</td>
			<td style="width:263px;">1201</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="121">
			<td height="121" style="height:121px;width:263px;">Narrow Pattern 12cm Curved Forceps</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">11003-12</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="141">
			<td height="141" style="height:141px;width:263px;">Curved Serated 10 cm Graefe Forceps&#160;</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">11052-10</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">Castroviejo Needle Holder</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">12565-14</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:263px;">Straight 9 cm Strabismus Scissors (Blunt Tip)</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">14075-09</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:263px;">Straight 8.5 cm Hardened Fine Scissors</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">14090-09</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:263px;">Straight, Blunt Tip Cohan-Vannas Spring Scissors</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">15000-12</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">Skin Glue</td>
			<td style="width:263px;">Gluture</td>
			<td style="width:263px;">32046</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">22 ga angiocatheter</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">24 ga angiocatheter</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">3 mL luer lock syringe</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">4 short retractors</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">2 long retractors</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:263px;">5-0 Prolene on curved cutting needle</td>
			<td style="width:263px;">Ethicon</td>
			<td style="width:263px;">8698G</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">0.5 mL syringe on 27 ga needle</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">Normal Saline</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">Buprenorphine</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
		</tr>
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mouse pneumonectomy model of compensatory lung growth

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to disability or death. Injury-based animal models of lung repair and regeneration are limited by injury-specific responses making it difficult to differentiate changes related to the injury response and injury resolution from changes related to lung repair and lung regeneration. However, use of animal models to identify these repair and regeneration signaling pathways is critical to the development of new therapies aimed at improving pulmonary function following lung injury. The mouse pneumonectomy model utilizes compensatory lung growth to isolate those repair and regeneration signals in order to more clearly define mechanisms of alveolar re-septation. Here, we describe our technique for performing mouse pneumonectomy and sham pneumonectomy. This technique may be utilized in conjunction with lineage tracing or other transgenic mouse models to define molecular and cellular mechanism of lung repair and regeneration.

. Une parcelle de PNX et le poids de souris subi une opération fictive est fourni à la figure 6 Dans nos mains, la survie est toujours de 95 à 100% pour les deux PNX et imposture pneumonectomie. Pour une description de la façon dont le poumon droit re-pousse dans ce modèle et l&#39;évolution dans le temps prévu, nous renvoyons le lecteur aux manuscrits de Gibney et al. 15 et Wang et al. 14 Plusieurs écueils les plus courants do...

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.

Souris Pneumonectomie modèle de croissance compensatoire Lung

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

12.7K vues.  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.

Vidéo: Mouse Pneumonectomy Model of Compensatory Lung Growth

A Simple Method of Mouse Lung Intubation

A simple procedure to intubate mice for pulmonary function measurements would have several advantages in longitudinal studies with limited numbers or expensive animal. One of the reasons that this is not done more routinely is that it is relatively difficult, despite there being several published studies that describe ways to achieve it. In this paper we demonstrate a procedure that eliminates one of the major hurdles associated with this intubation, that of visualizing the trachea during the entire time of intubation. The approach uses a 0.5 mm fiberoptic light source that serves as an introducer to direct the intubation cannula into the mouse trachea. We show that it is possible to use this procedure to measure lung mechanics in individual mice over a time course of at least several weeks. The technique can be set up with relatively little expense and expertise, and it can be routinely accomplished with relatively little training. This should make it possible for any laboratory to routinely carry out this intubation, thereby allowing longitudinal studies in individual mice, thereby minimizing the number of mice needed and increasing the statistical power by using each mouse as its own control.

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

Une méthode simple d&#39;intubation poumon de souris

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

Recherche

Médecine

Diagnostic Ultrasound Imaging of Mouse Diaphragm Function

Function analysis of rodent respiratory skeletal muscles, particularly the diaphragm, is commonly performed by isolating muscle strips using invasive surgical procedures. Although this is an effective method of assessing in vitro diaphragm activity, it involves non-survival surgery. The application of non-invasive ultrasound imaging as an in vivo procedure is beneficial since it not only reduces the number of animals sacrificed, but is also suitable for monitoring disease progression in live mice. Thus, our ultrasound imaging method may likely assist in the development of novel therapies that alleviate muscle injury induced by various respiratory diseases. Particularly, in clinical diagnoses of obstructive lung diseases, ultrasound imaging has the potential to be used in conjunction with other standard tests to detect the early onset of diaphragm muscle fatigue. In the current protocol, we describe how to accurately evaluate diaphragm contractility in a mouse model using a diagnostic ultrasound imaging technique.

Diagnostic ultrasound imaging has proven to be effective in diagnosing various respiratory diseases in human and animal subjects. We demonstrate a comprehensive ultrasound protocol utilized by Dr. Zuo's lab to analyze diaphragm kinetics specifically in mouse models. This is also a non-invasive research technique which can provide quantitative information on mouse respiratory muscle function.

L&#39;échographie diagnostique de souris membrane Fonction

Function analysis of rodent respiratory skeletal muscles, particularly the diaphragm, is commonly performed by isolating muscle strips using invasive ...

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms at work during MI and developing effective therapeutic approaches requires methodology to reproducibly simulate the clinical incidence and reflect the pathophysiological changes associated with MI. Here, we describe a surgical method to induce MI in mouse models that can be used for short-term ischemia-reperfusion (I/R) injury as well as permanent ligation. The major advantage of this method is to facilitate location of the left anterior descending artery (LAD) to allow for accurate ligation of this artery to induce ischemia in the left ventricle of the mouse heart. Accurate positioning of the ligature on the LAD increases reproducibility of infarct size and thus produces more reliable results. Greater precision in placement of the ligature will improve the standard surgical approaches to simulate MI in mice, thus reducing the number of experimental animals necessary for statistically relevant studies and improving our understanding of the mechanisms producing cardiac dysfunction following MI. This mouse model of MI is also useful for the preclinical testing of treatments targeting myocardial damage following MI.

We introduce a surgical method to induce experimental ischemia/reperfusion (I/R) injury to simulate myocardial infarction (MI) in mouse models that allows for more clarity in positioning of the ligation on the left anterior descending artery (LAD) to increase the reproducibility of MI experiments in mice.

Un modèle murin d&#39;ischémie myocardique-reperfusion par ligature de l&#39;artère interventriculaire antérieure

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms ...

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.

Implantation de fibrine Gel sur la souris Lung pour étudier l&#39;angiogenèse pulmonaire spécifique

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

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.

Méthode de isolé<em&gt; Ex Vivo</em&gt; Lung perfusion dans un modèle rat: Leçons tirées élaboration d&#39;un programme EVLP Rat

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

Real-time X-ray Imaging of Lung Fluid Volumes in Neonatal Mouse Lung

At birth, the lung undergoes a profound phenotypic switch from secretion to absorption, which allows for adaptation to breathing independently. Promoting and sustaining this phenotype is critically important in normal alveolar growth and gas exchange throughout life. Several in vitro studies have characterized the role of key regulatory proteins, signaling molecules, and steroid hormones that can influence the rate of lung fluid clearance. However, in vivo examinations must be performed to evaluate whether these regulatory factors play important physiological roles in regulating perinatal lung liquid absorption. As such, the utilization of real time X-ray imaging to determine perinatal lung fluid clearance, or pulmonary edema, represents a technological advancement in the field. Herein, we explain and illustrate an approach to assess the rate of alveolar lung fluid clearance and alveolar flooding in C57BL/6 mice at post natal day 10 using X-ray imaging and analysis. Successful implementation of this protocol requires prior approval from institutional animal care and use committees (IACUC), an in vivo small animal X-ray imaging system, and compatible molecular imaging software.

We present a protocol to assess the rate of alveolar fluid clearance or pulmonary edema in neonatal mouse lung using X-ray imaging technology.

En temps réel d&#39;imagerie à rayons X du poumon Volumes fluide dans néonatale Souris Lung

At birth, the lung undergoes a profound phenotypic switch from secretion to absorption, which allows for adaptation to breathing independently. Promoting ...

Apical Resection Mouse Model to Study Early Mammalian Heart Regeneration

Cardiovascular disease plagues the whole world due to intensive lifestyle changes. Heart regeneration holds great promise for repairing and restoring cardiomyocytes lost due to injury and disease. In contrast to the robust cardiac regeneration of certain lower vertebrates, adult mammalian hearts typically show minimal capacity for heart regeneration and repair. However, recent studies have sparked considerable scientific interest with the finding that, between postnatal day 1 to 7 (P1 to P7), the neonatal mouse heart retains significant regenerative potential after apical resection (i.e., surgical amputation and exposure of left ventricular apex). One major controversy over this finding might be due to the diverse surgery-related procedures used in efforts to replicate or expand upon this important finding. These instructions dynamically present the materials and methodology for apical resection in a mouse model. The salient steps of this rodent survival surgery involve hypothermia anesthesia, thoracotomy, surgical amputation of heart ventricular apex, and suture and recovery of mice. The approach described could expand the application of the apical resection mouse model for cardiovascular research.

A step-by-step video protocol of apical resection is demonstrated in this study. Apical resection is a recently highlighted surgical approach in mammalian heart regeneration research. This study may promote the application of apical resection as a standard methodology in research into the mechanism underlying heart regeneration.

Modèle apicale résection souris pour étude au début de régénération de coeur de mammifère

Cardiovascular disease plagues the whole world due to intensive lifestyle changes. Heart regeneration holds great promise for repairing and restoring ...

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute respiratory distress syndrome (ARDS), characterized by neutrophilic alveolitis, and injury to both alveolar epithelium and vascular endothelium. Clinically, ARDS is defined by acute onset of hypoxemia, bilateral patchy pulmonary infiltrates and non-cardiogenic pulmonary edema. Human studies have provided us with valuable information about the physiological and inflammatory changes in the lung caused by ARDS, which has led to various hypotheses about the underling mechanisms. Unfortunately, difficulties determining the etiology of ARDS, as well as a wide range of pathophysiology have resulted in a lack of critical information that could be useful in developing therapeutic strategies.
Translational animal models are valuable when their pathogenesis and pathophysiology accurately reproduce a concept proven in both in vitro and clinical settings. Although large animal models (e.g., sheep) share characteristics of the anatomy of human trachea-bronchial tree, murine models provide a host of other advantages including: low cost; short reproductive cycle lending itself to greater data acquisition; a well understood immunologic system; and a well characterized genome leading to the availability of a variety of gene deletion and transgenic strains. A robust model of low pH induced ARDS requires a murine ALI that targets mainly the alveolar epithelium, secondarily the vascular endothelium, as well as the small airways leading to the alveoli. Furthermore, a reproducible injury with wide differences between different injurious and non-injurious insults is important.
The murine gastric acid aspiration model presented here using hydrochloric acid employs an open tracheostomy and recreates a pathogenic scenario that reproduces the low pH pneumonitis injury in humans. Additionally, this model can be used to examine interaction of a low pH insult with other pulmonary injurious entities (e.g., food particles, pathogenic bacteria).

This protocol induces acute lung injury in a mouse that has close fidelity to the pathogenesis of acid pneumonitis observed in humans. We generate a maximal acute nonlethal low pH lung injury and account for differences in rodent-human anatomic respiratory structure using an open tracheostomy coupled with circumferential pressure release.

Ouvrir trachéotomie acide gastrique Aspiration modèle murin de lésion pulmonaire aiguë Résultats dans Maximal aiguë non létaux Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...

Integrated Compensatory Responses in a Human Model of Hemorrhage

Hemorrhage is the leading cause of trauma-related deaths, partly because early diagnosis of the severity of blood loss is difficult. Assessment of hemorrhaging patients is difficult because current clinical tools provide measures of vital signs that remain stable during the early stages of bleeding due to compensatory mechanisms. Consequently, there is a need to understand and measure the total integration of mechanisms that compensate for reduced circulating blood volume and how they change during ongoing progressive hemorrhage. The body&#39;s reserve to compensate for reduced circulating blood volume is called the &#39;compensatory reserve&#39;. The compensatory reserve can be accurately evaluated with real-time measurements of changes in the features of the arterial waveform measured with the use of a high-powered computer. Lower Body Negative Pressure (LBNP) has been shown to simulate many of the physiological responses in humans associated with hemorrhage, and is used to study the compensatory response to hemorrhage. The purpose of this study is to demonstrate how compensatory reserve is assessed during progressive reductions in central blood volume with LBNP as a simulation of hemorrhage.

The purpose of this protocol is to demonstrate the techniques for measuring compensatory responses to reduced central blood volume using lower body negative pressure as a noninvasive experimental model of human hemorrhage which can be used to quantify the total integration of compensatory mechanisms to blood volume deficit in humans.

Les réponses compensatoires intégrées dans un modèle humain de Hémorragie

Hemorrhage is the leading cause of trauma-related deaths, partly because early diagnosis of the severity of blood loss is difficult. Assessment of ...

Echocardiographic Measurement of Right Ventricular Diastolic Parameters in Mouse

Diastolic dysfunction is a prominent feature of right ventricular (RV) remodeling associated with conditions of pressure overload. However, the RV diastolic function is rarely quantified in experimental studies. This might be due to technical difficulties in the visualization of the RV in the apical four-chamber view in rodents. Here we describe two positions facilitating the visualization of the apical four-chamber view in mice to assess the RV diastolic function.
The apical four-chamber view is enabled by tilting the mouse fixation platform to the left and caudally (LeCa) or to the right and cranially (RiCr). Both positions provide images of comparable quality. The results of the RV diastolic function obtained from two positions are not significantly different. Both positions are comparably easy to perform. This protocol can be incorporated into published protocols and enables detailed investigations of the RV function.

Here we describe and compare two positions for obtaining the apical four-chamber view in mice. These positions enable the quantification of the right ventricular function, provide comparable results, and can be used interchangeably.