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.

HINWEIS: Tier Verwendung Aussage: Alle Verfahren in dieser Studie wurden mit Genehmigung und nach den Richtlinien der Institutional Animal Benutzung und Pflege Committee (IACUC) in Cincinnati Kinderklinik durchgeführt. Acht Wochen alte C57BL / 6J männliche Mäuse wurden von Jackson Laboratories (Bar Harbor, ME) erhalten und man ließ sie für eine Woche vor dem Gebrauch akklimatisieren. Bis der Operation wurden die Tiere in einem pathogenfreien Barriere Anlage untergebracht und zur Verfügung gestellt autoklaviert Chow und gefiltertes Wasser ad libdium. Jede Maus Käfig wurde mit einer dedizierten Luft und Wasser versorgt, und die Zimmer wurden auf einem 12 Stunden Tag-Nacht-Zyklus gehalten. Nach Erholung von der Operation wurden die Mäuse in Käfigen mit gefiltertem Spitzen aufrechterhalten, sofern autoklaviert Futter ad libidum und sofern aus einer Wasserflasche gefiltertes Wasser. 1. Herstellung des Instruments <ol><li> Stellen Sie 6 Haut Wundhaken mit Büroklammern und Pins. Twist bogene Büroklammern auf den Schäften der Papier-Pins, lTraufe eine 5 cm gerader Stahldraht an einem Ende und machen eine einzige 0,5 cm &quot;U&quot; förmigen Haken am Ende des Drahtes. </li><li> Stellen ca. 15 x 15 cm im Quadrat-OP-Abdeckungen mit Plastikfolie. Bereiten Sie eine Umkleide pro Maus. Setzen Sie ein Papierturm zwischen den einzelnen Wickel. </li><li> Sterilisieren Sie alle chirurgische Instrumente zusammen mit einem Stapel von 12 x 12 Zoll Korkplatten, Gaze, und Wattestäbchen. </li></ol> 2. Maus Vorbereitung <ol><li> Induzieren Anästhesie mit 2% Isofluran. Wiegen Tier. </li><li> In einem speziellen Operationsvorbereitungsbereich rasieren linken Brust und Nackenbereich mit elektrischen Rasierer. </li><li> Geben Sie einen Tropfen der künstliche Tränen Salbe die Augen die Maus. </li><li> Dekontaminieren Hals und linken Brust mit Chlorhexidin und Isopropylalkohol. Wiederholen Sie zweimal. </li></ol> 3. Maus-Oro-Intubation und Beatmung <ol><li> Haben Sie ein nicht-sterilen OP-Techniker statt mit der Maus auf dem Rücken in der vorgewärmten OP-Bereich.</li><li> Narkosetiefe Bestätigen Dokumentation Fehlen einer Reaktion auf die Pfote Prise. </li><li> Nach dem Waschen der Hände und das Anziehen OP-Kleidung, Maske und Hut, don sterile Handschuhe. </li><li> Nach Drapierungen und unter aseptischen Bedingungen, machen Sie einen 1 cm vertikale Inzision über der vorderen Mitte Nacken, den Kehlkopf freizulegen. Leicht einfahren Halsmuskulatur mit gebogenen, gezackten 10 cm Pinzette und setzen den Kehlkopf und Luftröhre durch die Streuung der Halsmuskulatur mit der Spitze einer geraden Schere. </li><li> Oral legen Sie eine 22 G stumpfen Spitze Angiokatheter in die Mitte der Luftröhre (1A) und optisch Platzierung (1B) zu bestätigen. Pflegen Anästhesie und belüften mit 1-3% Isofluran durch Nagetierventilator (225 ul pro Hub; 200 Stokes pro min). Verwenden Sie ein Druckgrenze von 15 cm H 2 O </li></ol> 4. Maus Pneumonektomie <ol><li> Legen Sie die Maus in die richtige Seitenlage mit der Maus den Rücken vorder Betreiber (links oben). Verwenden Sie einen selbstdichtenden Plastikfolie als sterilen Tuch. Schneiden durch den Faltenwurf, verwenden stumpfer Spitze gebogene Schere, einen 2 cm langen Schnitt parallel zu den Rippen auf der 4. und 5. Zwischenrippenraum zu machen. Legen Sie die stumpfe Spitze gebogene Schere und sezieren die Haut von den zugrunde liegenden Rippen und Rippenmuskeln. </li><li> Ziehen Sie die Haut mit vier Wundhaken, um ein Quadrat aussetzen 1,5 x 1,5 cm Operationsfenster (Abbildung 2A). Sichern Sie die Wundhaken an den Kork Bord. </li><li> Präparieren Sie auf Rippen mit einer gebogenen Pinzette, und verwenden Sie eine Spitze der gebogenen Pinzette, um die Brusthöhle zu gelangen. </li><li> Mit Hilfe der stumpfen Spitze Mikroscheren, verwenden Sie die Untermesser, um die Brusthöhle zu gelangen. Machen Sie eine 0,5 cm Einschnitt zwischen den Rippen, und wiederholen Sie in die entgegengesetzte Richtung. </li><li> Mit den beiden verbleibenden Wundhaken, öffnen Sie den Brustkorb in der Vorwärts-Rückwärts-Achse und die sichere die Retraktoren mit dem Kork Bord (2B). </li> &lt;li&gt; Mit gebogenen stumpfen Pinzette in der linken Hand das linke Lunge und verdrängen den oberen Teil der linken Lunge seitlich und unten durch die Thorakotomie, bis die linke Lungenarterie und Bronchien ausgesetzt sind (3A, B). </li><li> Halten Sie die geladenen Titan Gefäßmikroclip Applikator in die rechte Hand mit dem Körper des Applikators in der Handfläche und gebogene Spitze nach weg von der Handfläche (3C), schieben Sie die Applikatorspitze in den Brustkorb entlang der Krümmung der hinteren Seite des linken Lunge und Clip der linken Bronchus und Lungenschlagader (3D). </li><li> Entfernen Sie den Applikator aber halten Sie die linke Lunge eingefahren. Fassen Sie die stumpfe Spitze Mikro-Schere mit der rechten Hand und schneiden Sie die Bronchien und Lungenarterie distal der Clip und entfernen linken Lunge (3E). </li><li> Entfernen Sie die Rippenspreizer. </li><li> Mit den gebogenen stumpfen Pinzette zu kneifen bis 1 cm von der Haut schlechter als die incisiauf, sondern über dem Niveau der Membran und setzen Sie eine 24 G Angiokatheter durch die Haut und in die linke Brusthöhle (4A, B). </li><li> Verwenden Sie 5-0 Prolene Naht zu zwei Knopfnähten auf der 4. und 5. Rippen platzieren, um die Brusthöhle zu schließen. </li><li> Entfernen Sie die Haut Wundhaken. Verwenden Sie zwei Sätze von Pinzette, um die Haut entlang der Länge des Einschnitts nähern und kleben die Haut verschlossen. </li><li> Schließen Sie einen 3 ml Luer-Lock-Spritze mit dem Angiokatheter und entfernen Restluft durch leichten Ansaugen und Zurückziehen des Angiokatheter. </li><li> Kleben Sie den Halsschnitt geschlossen mit zwei Sätzen von Zangen wie zuvor. </li></ol> 5. Maus Sham Pneumonektomie <ol><li> Setzen Sie die linke Lunge wie in &quot;Maus Pneumonektomie&quot; Protokoll vermerkt. Heben Sie den Brustkorb mit gebogenen stumpfen Pinzette, um Luft in die linke Brusthöhle zu ermöglichen (5A, B). </li><li> Legen Sie eine 24 G Angiokatheter in die linke thoRacic Hohlraum wie oben darauf achten, daß die linke Lunge schädigen. </li><li> Mit 5-0 Prolene Nahtmaterial und darauf achten, daß die Lunge (5C) durchstoßen, indem Sie zwei Längen von Nahtmaterial in der 3. / 4 und 5. / 6. Rippenzwischenräumen (5D). Gebe beide Längen von Nahtmaterial vor den Bund, um die Risiken der linken Lunge Herniation zu verringern. Binden Sie die Nahtmaterial zu zwei unterbrochenen Stichen (5E) zu machen. </li><li> Kleben Sie die Haut über der Brust-Schnitt, entfernen Sie Restluft mit dem Angiokatheter und kleben Sie den Halsschnitt wie oben. </li></ol> 6. Wiederbelebung, Analgesie und Wiederherstellung <ol><li> Schalten Sie die Isofluran und Verwalten 0,1 mg / kg von Buprenorphin und 0,5 ml physiologischer Kochsalzlösung subkutan. </li><li> Wenn spontane Atmung wieder aufzunehmen, entfernen Sie den Endotrachealtubus. </li><li> Beachten Maus, bis es wieder ambulante ist. Wandern in der Regel Resum¯es mehrere Minuten nach der Entfernung des Endotrachealtubus. </li><li> Platzieren Sie die Maus in einem 27 ° C Inkubator (befeuchteten, Sauerstoff 25%) zu erholen O / N. HINWEIS: Legen Sie mehrere Pellets von Chow mit Wasser für die ersten 24 Stunden nach der Operation benetzt auf dem Käfigboden. </li><li> Verabreichen 0,1 mg / kg Buprenorphin durch intraperitoneale Injektion zweimal täglich für drei Tage nach der Operation. Darauf achten, dass die Operationsstelle beim Umgang mit Tieren zu öffnen. </li></ol> 7. Maus-Überwachung <ol><li> Wiegen Mäusen nach 1, 3, 5 und 7 Tage nach der Operation. </li></ol>

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The authors have nothing to disclose.

Wir haben die detaillierte Beschreibung der Maus PNX und Maus Schein PNX Verfahren bis heute berichtet vorgesehen. Wir haben die Leser bewusst einige der häufigsten Fallstricke, dass die Ermittler das Erlernen der häufigsten Verfahren stoßen gemacht, und wir haben verschiedene Techniken, die von unserem Labor entwickelt, um gegen diese Gefahren zu mildern skizziert. Andere Labors nutzen dieses Modell können andere Technik Modifikationen entwickelt haben, oder verwenden Sie verschiedene Instrumente. Bei der Bewertung...

Die Hauptfunktion der Lunge ist für Sauerstoff und Kohlendioxid Austausch zwischen einem Organismus und der Atmosphäre bereitzustellen. Beim Menschen eine Vielzahl von angeborenen und erworbenen Bedingungen führen zu verminderter Lungenfläche, die zu einer Beeinträchtigung der Lungenfunktion führt. Dh Lungen - Obwohl eine Vielzahl von Therapien wie inhalativen Kortikosteroiden, Bronchodilatatoren, zusätzlichen Sauerstoff und chronische mechanische Belüftung verwendet werden, um die Folgen der Beeinträchtigung der Lungenfunktion 1-3 abzuschwächen, wäre die ideale Therapie für diesen Bedingungen Nachwachsen der funktionellen Lungengewebe zu fördern Regeneration. Säugetiergeweberegeneration ist gut dokumentiert worden. Die Afrikanische Stachelmaus können große Bereiche der Haut ohne Narbenbildung 4 zu regenerieren. Die Phalanx beim Menschen nach einer Verletzung oder Amputation 5-7 regenerieren. Nach Pneumonektomie (PNX) tritt Ausgleichslungenwachstum bei Mäusen, 8, 9 Ratten, zu tungs 10 und 11 Menschen. Per Definition beinhaltet Ausgleichslungenwachstum nicht nur Ausbau der bestehenden Lufträume, aber erneut Septierung dieser vergrößerten Lufträume mit der Expansion des zugehörigen Mikrozirkulation 12. Genexpressionsanalyse hat gezeigt, dass dieses Modell rekapituliert viele der Signalwege der Lungenentwicklung 13. Vier Wochen nach der Maus PNX ist Alveolaroberfläche äquivalent zu der scheinoperierten Tieren 14. In diesem Manuskript beschreiben wir die Maus PNX und Schein PNX Verfahren.

<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">
			<td height="21" style="height:21px;width:263px;">Press-n-Seal Wrap</td>
			<td style="width:263px;">Glad Products Company</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.

. Ein Grundstück von PNX und scheinoperierten Maus Gewichte ist in Abbildung 6 vorgesehen In unseren Händen, ist das Überleben konsequent 95-100% für beide PNX und Schein Pneumonektomie. Eine Beschreibung, wie die rechte Lunge wieder wächst in diesem Modell und dem erwarteten zeitlichen Verlauf, verweisen wir den Leser auf Manuskripte Gibney et al. 15 und Wang et al. 14 Einige häufige Fehler zu vermeiden, um die Maus PNX und Maus...

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.

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 ...

mouse-pneumonectomy-model-of-compensatory-lung-growth

Research

JoVE Journal

Medicine

12.7K Aufrufe.  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.

Serie: 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.

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  ...

Forschung

Medizin

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.

Diagnostische Ultraschall-Bildgebung der Maus Membranfunktion

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.

Einem Mausmodell der myokardialen Ischämie-Reperfusion Injury durch Ligation des linken vorderen absteigenden Arterie

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.

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 ...

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 ...

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.

Echtzeit-Röntgenbildgebung von Lungenflüssigkeitsvolumen in Neonatal Mauslunge

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.

Wurzelspitzenresektion Mausmodell zur frühen Säugetierherzregeneration Studieren

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.

Offene Tracheotomie Magensäure Aspiration Mausmodell der akuten Lungenschädigung Ergebnisse in Maximal Akute 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 ...

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.

Integrierte kompensatorische Antworten in einem menschlichen Modell von Einblutung

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.