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.

Uso affermazione Animal:: NOTA Tutte le procedure di questo studio sono stati condotti con l&#39;approvazione e seguendo le linee guida del Istituzionale uso degli animali e del Comitato Care (IACUC) dell&#39;Ospedale dei bambini di Cincinnati. Otto settimane di età C57BL / 6J maschi sono stati ottenuti da Jackson Laboratories (Bar Harbor, ME) e permesso di ambientarsi per una settimana prima dell&#39;uso. Fino chirurgia, gli animali sono stati alloggiati in una struttura senza barriere-patogeno e fornito chow autoclavato e filtrata ad acqua libdium. Ogni gabbia mouse è stato alimentato con aria dedicato e acqua e sale sono stati mantenuti su una 12 hr ciclo giorno-notte. Dopo il recupero da un intervento chirurgico, i topi sono stati mantenuti in gabbie con cime filtrati, a condizione autoclavato chow ad libidum, e purché filtrata l&#39;acqua da una bottiglia d&#39;acqua. 1. Preparazione degli strumenti <ol><li> Rendere 6 divaricatori pelle utilizzando clip e perni di carta. Twist raddrizzò graffette sui gambi di perni di carta, lgronda un filo di acciaio diritto 5 cm da una estremità e fare un unico 0,5 centimetri gancio a forma di &quot;U&quot; alla fine del filo. </li><li> Fare circa 15 x 15 cm teli chirurgici quadrati con involucro di plastica. Preparare un condimento per ogni mouse. Mettere una torre di carta tra ogni involucro. </li><li> Sterilizzare tutti gli strumenti chirurgici con una pila di piastrelle 12 x 12 pollici sughero, garze e tamponi di cotone con punta. </li></ol> 2. Preparazione del mouse <ol><li> Indurre l&#39;anestesia con il 2% isoflurano. Pesare animale. </li><li> In un&#39;area dedicata, la preparazione chirurgica radersi torace sinistro e collo con il rasoio elettrico. </li><li> Applicare una goccia di lacrima artificiale unguento agli occhi del topo. </li><li> Decontaminare collo e torace sinistro con clorexidina e alcool isopropilico. Ripetere altre due volte. </li></ol> 3. mouse oro-tracheale intubazione e ventilazione meccanica <ol><li> Avere un luogo non sterile tecnico chirurgico la supina mouse nell&#39;area chirurgica pre-riscaldato.</li><li> Confermare profondità dell&#39;anestesia documentando la mancanza di una risposta a zampa pizzico. </li><li> Dopo aver lavato le mani e indossare abiti chirurgica, maschera e cappello, indossare guanti chirurgici sterili. </li><li> Dopo drappeggio e con tecnica asettica, fare una incisione verticale 1 centimetro sopra l&#39;anteriore a metà collo per esporre la laringe. Leggermente ritrarre i muscoli cinghia con curve dentata 10 centimetri pinze ed esporre la laringe e della trachea, diffondendo i muscoli cinghia con la punta di un paio di forbici diritte. </li><li> Via orale inserire un 22 G smussato punta angiocatheter nel mid-trachea (Figura 1A) e confermare visivamente il posizionamento (Figura 1B). Mantenere l&#39;anestesia e ventilare con 1-3% isoflurano attraverso roditori ventilatore (225 ml per ictus; 200 Stokes al minuto). Impiegare un limite pressione di 15 cm H 2 O. </li></ol> 4. mouse Pneumonectomia <ol><li> Posare il mouse in decubito laterale destro con la parte posteriore del topo di frontel&#39;operatore (lato sinistro in alto). Utilizzare un involucro di plastica autosigillante come un telino sterile. Taglio attraverso il drappo, utilizzare smussato forbici curve a punta per fare un taglio di due centimetri lungo parallelo alle costole al 4 ° e 5 ° spazio intercostale. Inserire la punta ricurva forbici spuntate e sezionare la pelle dalle costole sottostanti e muscoli intercostali. </li><li> Ritrarre la pelle con quattro divaricatori per esporre un quadrato di 1,5 x 1,5 centimetri finestra chirurgica (Figura 2A). Fissare i divaricatori alla scheda di sughero. </li><li> Sezionare fino a costine con pinze curve, e utilizzare una punta delle pinze curve di entrare nella cavità toracica. </li><li> Utilizzando la punta smussata micro-forbici, usare la lama inferiore per entrare nella cavità toracica. Eseguire un&#39;incisione 0,5 centimetri tra le costole e ripetere nella direzione opposta. </li><li> Utilizzando i due divaricatori rimanenti, aprire il torace nel asse anteriore-posteriore e la sicurezza dei divaricatori per la bacheca di sughero (Figura 2B). </li> &lt;li&gt; Uso curve pinze a punta smussata nella mano sinistra, afferrare il polmone sinistro e spostare la parte superiore del polmone sinistro lateralmente e inferiormente attraverso la toracotomia fino l&#39;arteria polmonare sinistra e dei bronchi sono esposti (Figura 3A, B). </li><li> Tenendo titanio applicatore microclip vascolare caricata nella mano destra con il corpo dell&#39;applicatore nel palmo e punta ricurva punta lontano dalla palma (Figura 3C), far scorrere la punta dell&#39;applicatore nel torace lungo la curvatura della faccia posteriore sinistra polmone e clip il bronco sinistro e l&#39;arteria polmonare (Figura 3D). </li><li> Rimuovere l&#39;applicatore ma mantenere il polmone sinistro retratto. Afferrare le smussati punta micro-forbici con la mano destra e tagliare il bronchi e polmonare distale al clip e rimuovere polmone sinistro (Figura 3E). </li><li> Rimuovere i divaricatori costali. </li><li> Utilizzare le pinze smussato curve a pizzicare 1 cm di pelle inferiori alla incisiacceso ma sopra del livello del diaframma e inserire un G angiocatheter 24 attraverso la pelle e nella cavità toracica sinistra (Figura 4A, B). </li><li> Utilizzare 5-0 prolene sutura di inserire due punti staccati in tutto il 4 ° e 5 ° costole per chiudere la cavità toracica. </li><li> Rimuovere i divaricatori pelle. Utilizzare due serie di pinze per approssimare la pelle lungo la lunghezza dell&#39;incisione e incollare la pelle chiusa. </li><li> Collegare una siringa luer-lock 3 ml al angiocatheter e rimuovere l&#39;aria residua applicando delicata aspirazione e il ritiro della angiocatheter. </li><li> Incollare l&#39;incisione collo chiuso con due set di pinze come prima. </li></ol> 5. mouse Sham Pneumonectomia <ol><li> Esporre il polmone sinistro, come indicato nel protocollo &quot;Mouse Pneumonectomia&quot;. Sollevare la gabbia toracica con una pinza smussato curve per consentire all&#39;aria nella cavità toracica sinistra (Figura 5A, B). </li><li> Posizionare un G angiocatheter 24 nel tho di sinistraRacic cavità come sopra facendo attenzione a non danneggiare il polmone sinistro. </li><li> Utilizzando 5-0 prolene sutura e facendo attenzione a non forare il polmone (figura 5C), posizionare due lunghezze di materiale di sutura nel 3 ° / 4 ° e 5 ° / 6 ° intercapedini costola (Figura 5D). Mettere entrambe le lunghezze di materiale di sutura prima di legare per ridurre i rischi di ernia del polmone sinistro. Legare il materiale di sutura per fare due punti interrotti (Figura 5E). </li><li> Incollare la pelle sopra l&#39;incisione toracica, rimuovere l&#39;aria residua con il angiocatheter, e incollare l&#39;incisione collo come sopra. </li></ol> 6. Rianimazione, Analgesia, e recupero <ol><li> Spegnere il isoflurano, e somministrare 0,1 mg / kg di buprenorfina e 0,5 ml di soluzione fisiologica normale per via sottocutanea. </li><li> Quando respirazioni spontanee riprendono, rimuovere il tubo endotracheale. </li><li> Osservare il mouse fino a quando non è di nuovo ambulatoriale. Walking tipicamente resumes diversi minuti dopo la rimozione del tubo endotracheale. </li><li> Posizionare il mouse in un 27 ° C incubatore (umidificato, 25% di ossigeno) per recuperare O / N. NOTA: Abbiamo posto diverse pellets di chow bagnato con acqua sul pavimento della gabbia per le prime 24 ore dopo l&#39;intervento chirurgico. </li><li> Somministrare 0,1 mg / kg di buprenorfina per via intraperitoneale due volte al giorno per tre giorni dopo l&#39;intervento. Fare attenzione a non aprire il sito chirurgico durante la manipolazione degli animali. </li></ol> 7. Monitoraggio mouse <ol><li> Pesare topi a 1, 3, 5, e 7 giorni dopo l&#39;intervento chirurgico. </li></ol>

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The authors have nothing to disclose.

Abbiamo fornito la descrizione più dettagliata delle procedure PNX del mouse PNX e mouse farsa segnalati fino ad oggi. Abbiamo fatto il lettore a conoscenza di alcuni dei problemi comuni che gli investigatori di apprendimento la procedura comunemente incontrano, e abbiamo delineato diverse tecniche sviluppate dal nostro laboratorio per mitigare queste trappole. Altri laboratori che utilizzano questo modello possono sviluppare altre modifiche tecnica o utilizzare strumenti diversi. Nel valutare le differenze nelle tecni...

La funzione principale del polmone è di fornire ossigeno e anidride carbonica scambio tra un organismo e l&#39;atmosfera. Negli esseri umani, una serie di condizioni congenite ed acquisite portano a ridurre la superficie del polmone che si traduce in deficit della funzionalità respiratoria. Anche se una serie di terapie come corticosteroidi inalatori, broncodilatatori, ossigeno supplementare, e la ventilazione meccanica cronica sono utilizzati per attenuare le conseguenze di funzionalità polmonare ridotta 1-3, la terapia ideale per queste condizioni dovrebbe promuovere la ricrescita del tessuto polmonare funzionale - cioè, polmone rigenerazione. Rigenerazione dei tessuti dei mammiferi è stata ben documentata. L&#39;africano Mouse spinosi può rigenerare ampie aree di pelle senza formazione di cicatrici 4. La falange distale nell&#39;uomo può rigenerarsi dopo una lesione o amputazione 5-7. Dopo pneumonectomia (PNX), la crescita del polmone di compensazione avviene nei topi 8, ratti 9, dogs 10, e gli esseri umani 11. Per definizione, la crescita del polmone di compensazione coinvolge non solo l&#39;espansione di spazi aerei esistenti, ma ri-septation di questi spazi aerei allargata con l&#39;espansione del microcircolo associato 12. Analisi di espressione genica ha dimostrato che questo modello ricapitola molti degli eventi di segnalazione dello sviluppo polmonare 13. Quattro settimane dopo il mouse PNX, superficie alveolare è equivalente a quella di fittizie animali operati 14. In questo manoscritto, descriviamo la PNX mouse e procedure PNX fittizi.

<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>
		<tr height="21">
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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.

. Una trama di PNX e sham gestito pesi del mouse viene fornito in figura 6 Nelle nostre mani, la sopravvivenza è costantemente 95-100% per entrambi PNX e sham pneumonectomia. Per una descrizione di come il polmone destro re-cresce in questo modello e il decorso previsto, rimandiamo il lettore di manoscritti di Gibney et al. 15 e Wang et al. 14 Diversi problemi comuni devono essere evitati per eseguire con successo le procedure sham pn...

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.

Topo Pneumonectomia Modello di crescita compensatoria 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 Views.  Cincinnati Children's Hospital Medical Center. Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.

Video: Mouse Pneumonectomy Model of Compensatory Lung Growth

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.

Un metodo semplice di intubazione mouse polmone

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

Ricerca

Medicina

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.

Diagnostic Ultrasound Imaging di funzione mouse Diaframma

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 modello murino di ischemia miocardica e riperfusione lesioni da legatura della arteria discendente anteriore sinistra

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.

L&#39;impianto di fibrina gel sul mouse polmone per studiare angiogenesi Lung-specifica

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.

Metodo di isolato<em&gt; Ex Vivo</em&gt; Lung Perfusion in un modello di ratto: Lezioni apprese da Sviluppo di un programma 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.

In tempo reale a raggi X Imaging del polmone volumi di fluido in neonatale topo polmone

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.

Modello apicale resezione mouse per primo studio Mammalian rigenerazione cardiaca

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.

Aprire tracheostomia gastrico acido aspirazione Modello murino di lesioni polmonari acute Risultati in massima acuta non letale 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.

Le risposte compensative integrati in un modello umano di emorragia

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.