Name: pre-clinical model of cardiac donation after circulatory death
Uploaded: 2019-08-02T13:30:00
Description: Watch this Scientific Journal Video about Pre-clinical Model of Cardiac Donation after Circulatory Death at JoVE.com

Une partie de ce travail a été soutenue par une généreuse contribution de la Fondation Marcel et Rolande Gosselin et de la Fondation M. Stefane Foumy. Nicolas Noiseux est chercheur au FRQ-S.
Les auteurs remercient Josh Zhuo Le Huang, Gabrielle Gascon, Sophia Ghiassi et Catherine Scalabrini pour leur soutien dans la collecte de données.

Tous les protocoles de soins et d'expérimentation des animaux conformes au Guide pour le soin et l'utilisation des animaux de laboratoire ont été approuvés par le comité institutionnel de soins et d'utilisation des animaux du Centre hospitalier de l'Université de Montréal.
1. Préparations préliminaires
<ol><li>Allumez le bain d'eau pour chauffer le système de livraison de cardioplégie (Figure 1A) et le système de perfusion ex vivo Langendorff (<stron...

<ol>
	<li>Gass, A. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+Transplantation+in+the+New+Era.">Cardiac Transplantation in the New Era.</a> Cardiology in Review. 23 (4), 182-188 (2015).</li><li>von Dossow, V., Costa, J., D'Ovidio, F., Marczin, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Worldwide+trends+in+heart+and+lung+transplantation:+Guarding+the+most+precious+gift+ever.">Worldwide trends in heart and lung transplantation: Guarding the most precious gift ever.</a> Best Practice &amp; Research. Clinical Anaesthesiology. 31 (2), 141-152 (2017).</li><li>Hornby, K., Ross, H., Keshavjee, S., Rao, V., Shemie, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-utilization+of+hearts+and+lungs+after+consent+for+donation:+a+Canadian+multicentre+study.">Non-utilization of hearts and lungs after consent for donation: a Canadian multicentre study.</a> Canadian Journal Of Anaesthesia. 53 (8), 831-837 (2006).</li><li>Manyalich, M., Nelson, H., Delmonico, F. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+need+and+opportunity+for+donation+after+circulatory+death+worldwide.">The need and opportunity for donation after circulatory death worldwide.</a> Current Opinion In Organ Transplantation. 23 (1), 136-141 (2018).</li><li>Shemie, S. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=National+recommendations+for+donation+after+cardiocirculatory+death+in+Canada:+Donation+after+cardiocirculatory+death+in+Canada.">National recommendations for donation after cardiocirculatory death in Canada: Donation after cardiocirculatory death in Canada.</a> CMAJ : Canadian Medical Association Journal. 175 (8), S1 (2006).</li><li>Page, A., Messer, S., Large, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+transplantation+from+donation+after+circulatory+determined+death.">Heart transplantation from donation after circulatory determined death.</a> Annals of Cardiothoracic Surgery. 7 (1), 75-81 (2018).</li><li>Monteagudo Vela, M., Garcia Saez, D., Simon, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+approaches+in+retrieval+and+heart+preservation.">Current approaches in retrieval and heart preservation.</a> Annals of Cardiothoracic Surgery. 7 (1), 67-74 (2018).</li><li>Dhital, K. K., Chew, H. C., Macdonald, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Donation+after+circulatory+death+heart+transplantation.">Donation after circulatory death heart transplantation.</a> Current Opinion In Organ Transplantation. 22 (3), 189-197 (2017).</li><li>McNally, S. J., Harrison, E. M., Wigmore, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ethical+considerations+in+the+application+of+preconditioning+to+solid+organ+transplantation.">Ethical considerations in the application of preconditioning to solid organ transplantation.</a> Journal of Medical Ethics. 31 (11), 631-634 (2005).</li><li>Rao, V., Feindel, C. M., Weisel, R. D., Boylen, P., Cohen, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Donor+blood+perfusion+improves+myocardial+recovery+after+heart+transplantation.">Donor blood perfusion improves myocardial recovery after heart transplantation.</a> The Journal of Heart and Lung Transplantation. 16 (6), 667-673 (1997).</li><li>Ramzy, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+allograft+preservation+using+donor-shed+blood+supplemented+with+L-arginine.">Cardiac allograft preservation using donor-shed blood supplemented with L-arginine.</a> The Journal of Heart and Lung Transplantation. 24 (10), 1665-1672 (2005).</li><li>Xin, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+New+Multi-Mode+Perfusion+System+for+Ex+vivo+Heart+Perfusion+Study.">A New Multi-Mode Perfusion System for Ex vivo Heart Perfusion Study.</a> Journal of Medical Systems. 42 (2), 25 (2017).</li><li>Messer, S., Ardehali, A., Tsui, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Normothermic+donor+heart+perfusion:+current+clinical+experience+and+the+future.">Normothermic donor heart perfusion: current clinical experience and the future.</a> Transplant International. 28 (6), 634-642 (2015).</li><li>Flecknell, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Laboratory Animal Anaesthesia (Fourth Edition). , 77-108 (2016).</li><li>Kearns, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Rodent+Model+of+Cardiac+Donation+After+Circulatory+Death+and+Novel+Biomarkers+of+Cardiac+Viability+During+Ex+vivo+Heart+Perfusion.">A Rodent Model of Cardiac Donation After Circulatory Death and Novel Biomarkers of Cardiac Viability During Ex vivo Heart Perfusion.</a> Transplantation. 101 (8), e231-e239 (2017).</li><li>Sandha, J. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Steroids+Limit+Myocardial+Edema+During+Ex+vivo+Perfusion+of+Hearts+Donated+After+Circulatory+Death.">Steroids Limit Myocardial Edema During Ex vivo Perfusion of Hearts Donated After Circulatory Death.</a> The Annals of Thoracic Surgery. 105 (6), 1763-1770 (2018).</li><li>Iyer, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increasing+the+tolerance+of+DCD+hearts+to+warm+ischemia+by+pharmacological+postconditioning.">Increasing the tolerance of DCD hearts to warm ischemia by pharmacological postconditioning.</a> American Journal of Transplantation. 14 (8), 1744-1752 (2014).</li><li>Sanz, M. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardioprotective+reperfusion+strategies+differentially+affect+mitochondria:studies+in+an+isolated+rat+heart+model+of+donation+after+circulatory+death+(DCD).">Cardioprotective reperfusion strategies differentially affect mitochondria:studies in an isolated rat heart model of donation after circulatory death (DCD).</a> American Journal of Transplantation. , (2018).</li><li>Van de Wauwer, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mode+of+death+in+the+non-heart-beating+donor+has+an+impact+on+lung+graft+quality.">The mode of death in the non-heart-beating donor has an impact on lung graft quality.</a> European Journal of Cardio-Thoracic Surgery. 36 (5), 919-926 (2009).</li><li>Quader, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+Optimal+Coronary+Flow+for+the+Preservation+of+&amp;#34;Donation+after+Circulatory+Death&amp;#34;+in+Murine+Heart+Model.">Determination of Optimal Coronary Flow for the Preservation of &amp;#34;Donation after Circulatory Death&amp;#34; in Murine Heart Model.</a> ASAIO journal (American Society for Artificial Internal Organs : 1992). 64 (2), 225-231 (2018).</li><li>Priebe, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+acute+open-chest+model.">The acute open-chest model.</a> British Journal Of Anaesthesia. 60 (8 Suppl 1), 38-41 (1988).</li><li>Narita, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+effects+of+vecuronium+and+its+interaction+with+autonomic+nervous+system+in+isolated+perfused+canine+hearts.">Cardiac effects of vecuronium and its interaction with autonomic nervous system in isolated perfused canine hearts.</a> Journal of Cardiovascular Pharmacology. 19 (6), 1000-1008 (1992).</li><li>Dhital, K. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adult+heart+transplantation+with+distant+procurement+and+ex-vivo+preservation+of+donor+hearts+after+circulatory+death:+a+case+series.">Adult heart transplantation with distant procurement and ex-vivo preservation of donor hearts after circulatory death: a case series.</a> Lancet (London, England). 385 (9987), 2585-2591 (2015).</li><li>Messer, S. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+assessment+and+transplantation+of+the+donor+heart+after+circulatory+death.">Functional assessment and transplantation of the donor heart after circulatory death.</a> The Journal of Heart and Lung Transplantation. 35 (12), 1443-1452 (2016).</li><li>White, C. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+donor+heart+viability+during+ex+vivo+heart+perfusion.">Assessment of donor heart viability during ex vivo heart perfusion.</a> Canadian Journal of Physiology and Pharmacology. 93 (10), 893-901 (2015).</li><li>Mayr, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+troponin+T+and+creatine+kinase+predict+mid-term+infarct+size+and+left+ventricular+function+after+acute+myocardial+infarction:+a+cardiac+MR+study.">Cardiac troponin T and creatine kinase predict mid-term infarct size and left ventricular function after acute myocardial infarction: a cardiac MR study.</a> Journal of Magnetic Resonance Imaging. 33 (4), 847-854 (2011).</li><li>Remppis, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracellular+compartmentation+of+troponin+T:+release+kinetics+after+global+ischemia+and+calcium+paradox+in+the+isolated+perfused+rat+heart.">Intracellular compartmentation of troponin T: release kinetics after global ischemia and calcium paradox in the isolated perfused rat heart.</a> Journal of Molecular and Cellular Cardiology. 27 (2), 793-803 (1995).</li><li>Rossello, X., Hall, A. R., Bell, R. M., Yellon, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+Langendorff+Perfused+Isolated+Mouse+Heart+Model+of+Global+Ischemia-Reperfusion+Injury:+Impact+of+Ischemia+and+Reperfusion+Length+on+Infarct+Size+and+LDH+Release.">Characterization of the Langendorff Perfused Isolated Mouse Heart Model of Global Ischemia-Reperfusion Injury: Impact of Ischemia and Reperfusion Length on Infarct Size and LDH Release.</a> Journal of Cardiovascular Pharmacology and Therapeutics. 21 (3), 286-295 (2016).</li><li>Dornbierer, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+reperfusion+hemodynamics+predict+recovery+in+rat+hearts:+a+potential+approach+towards+evaluating+cardiac+grafts+from+non-heart-beating+donors.">Early reperfusion hemodynamics predict recovery in rat hearts: a potential approach towards evaluating cardiac grafts from non-heart-beating donors.</a> PloS One. 7 (8), e43642 (2012).</li><li>Henry, P. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Positive+staircase+effect+in+the+rat+heart.">Positive staircase effect in the rat heart.</a> The American Journal of Physiology. 228 (2), 360-364 (1975).</li><li>Markert, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+a+method+to+correct+the+contractility+index+LVdP/dt(max)+for+changes+in+heart+rate.">Evaluation of a method to correct the contractility index LVdP/dt(max) for changes in heart rate.</a> Journal of Pharmacological and Toxicological Methods. 66 (2), 98-105 (2012).</li><li>Azar, T., Sharp, J., Lawson, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+rates+of+male+and+female+Sprague-Dawley+and+spontaneously+hypertensive+rats+housed+singly+or+in+groups.">Heart rates of male and female Sprague-Dawley and spontaneously hypertensive rats housed singly or in groups.</a> Journal of the American Association for Laboratory Animal Science. 50 (2), 175-184 (2011).</li><li>Bonney, S., Hughes, K., Eckle, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anesthetic+cardioprotection:+the+role+of+adenosine.">Anesthetic cardioprotection: the role of adenosine.</a> Current Pharmaceutical Design. 20 (36), 5690-5695 (2014).</li><li>Ali, A. 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Le protocole présenté ici introduit un modèle simple, pratique et polyvalent de DCD cardiaque, offrant la possibilité d'évaluer la récupération fonctionnelle cardiaque, les dommages de tissu et l'utilisation des agents cardioprotecteurs de poteau-conditionnement pour améliorer le rétablissement du donneur cœurs autrement jetés pour la transplantation. Les systèmes de perfusion cardiaque ex vivo (EVHP) ont été optimisés pour fournir une plate-forme pour évaluer la fonction cardiaque et offrir une occasion ...

La transplantation d'organes solides en général et la transplantation cardiaque, en particulier, sont à la hausse dans le monde1,2. La méthode standard d'achat d'organes est le don après la mort cérébrale (DBD). Compte tenu des critères d'inclusion stricts de la DBD, moins de 40% des cœurs offerts sont acceptés3, limitant ainsi l'offre face à la demande croissante et l'extension de la liste d'attente des organes. Pour résoudre ce problème, l'utilisation d'organes donnés après la mort circulatoire (DCD) est considérée comme une solution potentielle4.
Chez les donneurs de DCD, cependant, une phase angoissée après le retrait des soins et une période d'ischémie chaude non protégée avant la réanimation sont inévitables5. Les dommages potentiels d'organe après la mort circulatoire peuvent mener au dysfonctionnement d'organe, expliquant la réticence à adopter systématiquement des transplantations de coeur de DCD. Il est rapporté que seulement 4 centres utilisent des coeurs de DCD médicalement, avecdes critères stricts qui incluent les temps chauds très courts d'ischémie chaude et les jeunes donateurs sans pathologies chroniques 6,7. Pour des raisons éthiques et juridiques, des interventions cardioprotectrices limitées ou inadaptées peuvent être appliquées chez les donneurs avant la mort circulatoire5,8,9. Ainsi, toute atténuation pour soulager les lésions ischémique-réperfusion (IR) est limitée aux thérapies cardioprotectrices initiées lors d'une réperfusion précoce avec des solutions cardioplégiques, et ne permettent pas une évaluation fonctionnelle appropriée. Perfusion cardiaque ex vivo (EVHP) et le reconditionnement du cœur DCD à l'aide de plates-formes dédiées a été proposé comme une solution alternative et étudié par divers chercheurs10,11,12,13 . EVHP offre une occasion unique de fournir des agents post-conditionnement aux cœurs DCD pour améliorer la récupération fonctionnelle. Cependant, pour une traduction clinique efficace, de nombreuses questions techniques et pratiques restent à résoudre, et cela est encore aggravé par l'absence de consensus sur une gamme de perfusion et de critères fonctionnels pour déterminer la transplantation6, 8.
Ici nous rapportons le développement d'un protocole préclinique reproductible de DCD d'animal de petit animal combiné avec un système ex vivo de perfusion de coeur qui peut être employé pour étudier le post-conditionnement d'organe lancé au moment de l'approvisionnement, pendant la reperfusion initiale, et /ou dans l'ensemble de l'EVHP.

<table>
	<tbody>
		<tr>
			<td>0,9% Sodium Chloride. 1L bag</td>
			<td>Baxter</td>
			<td></td>
			<td>Electrolyte solution for flushing in the modified Langendorff system.</td>
		</tr>
		<tr>
			<td>14G 2&#34; I.V catheter</td>
			<td>Jelco</td>
			<td>4098</td>
			<td>To act as endotracheal tube.</td>
		</tr>
		<tr>
			<td>2,3,5-Triphenyltetrazolium chloride</td>
			<td>Milipore-Sigma</td>
			<td>T8877</td>
			<td>Vital coloration</td>
		</tr>
		<tr>
			<td>22G 1&#34; I.V catheter</td>
			<td>BD</td>
			<td>383532</td>
			<td>I.V catheter with extension tube that facilitates manipulation for carotid catheterization</td>
		</tr>
		<tr>
			<td>Adson Dressing Fcp, 4 3/4&#34;, Serr</td>
			<td>Skalar</td>
			<td>50-3147</td>
			<td>Additional forceps for tissue manipulation</td>
		</tr>
		<tr>
			<td>Alm Self-retaining retractor 4x4 Teeth Blunt 2-3/4&#34;</td>
			<td>Skalar</td>
			<td>22-9027</td>
			<td>Tissue retractor used to maintain the chest open.</td>
		</tr>
		<tr>
			<td>Bridge amp</td>
			<td>ADinstruments</td>
			<td>FE221</td>
			<td>Bridge amp for intracarotid blood pressure measurement</td>
		</tr>
		<tr>
			<td>Calcium chloride</td>
			<td>Milipore-Sigma</td>
			<td>C1016</td>
			<td>CaCl2 anhydrous, granular, &#8804;7.0 mm, &#8805;93.0% Part of the Krebs solution</td>
		</tr>
		<tr>
			<td>D-(+)-Glucose</td>
			<td>Milipore-Sigma</td>
			<td>G8270</td>
			<td>D-Glucose &#8805;99.5% Part of the Krebs solution</td>
		</tr>
		<tr>
			<td>DIN(8) to Disposable BP Transducer</td>
			<td>ADinstruments</td>
			<td>MLAC06</td>
			<td>Adapter cable for link between bridge amp and pressure transducer</td>
		</tr>
		<tr>
			<td>Disposable BP Transducer (stopcock)</td>
			<td>ADinstruments</td>
			<td>MLT0670</td>
			<td>Pressure transducer for intracarotid blood pressure measurement</td>
		</tr>
		<tr>
			<td>dPBS</td>
			<td>Gibco</td>
			<td>14190-144</td>
			<td>Electrolyte solution without calcium or magnesium.</td>
		</tr>
		<tr>
			<td>Eye Dressing Fcp, Str, Serr, 4&#34;</td>
			<td>Skalar</td>
			<td>66-2740</td>
			<td>Additional forceps for tissue manipulation</td>
		</tr>
		<tr>
			<td>Formalin solution, neutral buffered, 10%</td>
			<td>Milipore-Sigma</td>
			<td>HT501128</td>
			<td>Fixative solution</td>
		</tr>
		<tr>
			<td>Heating Pad</td>
			<td>Sunbean</td>
			<td>756-CN</td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin sodium 1000 UI/mL</td>
			<td>Sandoz</td>
			<td></td>
			<td>For systemic anticoagulation</td>
		</tr>
		<tr>
			<td>Hydrochloric Acid 36,5 to 38,0%</td>
			<td>Fisher scientific</td>
			<td>A144-500</td>
			<td>Diluted 1:1 for pH correction</td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Bimeda</td>
			<td></td>
			<td>Anesthetic. 100 mg/ml</td>
		</tr>
		<tr>
			<td>LabChart</td>
			<td>ADinstruments</td>
			<td></td>
			<td>Control software for the Powerlab polygraph, allowing off-line analyses. Version 7, with blood pressure and PV loop modules enabled</td>
		</tr>
		<tr>
			<td>Left ventricle pressure balloon</td>
			<td>Radnoti</td>
			<td>170404</td>
			<td>In latex. Size 4.</td>
		</tr>
		<tr>
			<td>Lidocaine HCl 2% solution</td>
			<td>AstraZeneca</td>
			<td></td>
			<td>Antiarrhythmic for the cardioplegic solution</td>
		</tr>
		<tr>
			<td>Magnesium Chloride ACS</td>
			<td>ACP Chemicals</td>
			<td>M-0460</td>
			<td>MgCl2+6H2O &#8805;99.0% Part of the Krebs solution</td>
		</tr>
		<tr>
			<td>Micro pressure sensor</td>
			<td>Radnoti</td>
			<td>159905</td>
			<td>Micro pressure sensor and amplifier connected to the intraventricular balloon</td>
		</tr>
		<tr>
			<td>Pacemaker</td>
			<td>Biotronik</td>
			<td>Reliaty</td>
			<td>Set to generate a pulse each 200 ms for a heart rate of 300 bpm.</td>
		</tr>
		<tr>
			<td>pH bench top meter</td>
			<td>Fisher scientific</td>
			<td>AE150</td>
			<td></td>
		</tr>
		<tr>
			<td>Physiological monitor</td>
			<td>Kent Scientific</td>
			<td>Physiosuite</td>
			<td>For continuous monitoring of rodent temperature and saturation during the procedure</td>
		</tr>
		<tr>
			<td>Plasma-Lyte A</td>
			<td>Baxter</td>
			<td></td>
			<td>Electrolyte solution used as base to prepare cardioplegia</td>
		</tr>
		<tr>
			<td>Potassium Chloride</td>
			<td>Milipore-Sigma</td>
			<td>P4504</td>
			<td>KCl &#8805;99.0% Part of the Krebs solution</td>
		</tr>
		<tr>
			<td>Potassium Chloride 2 meq/ml</td>
			<td>Hospira</td>
			<td></td>
			<td>Part of the cardioplegic solution</td>
		</tr>
		<tr>
			<td>PowerLab 8/30 Polygraph</td>
			<td>ADinstruments</td>
			<td></td>
			<td>Electronic polygraph</td>
		</tr>
		<tr>
			<td>Silk 2-0</td>
			<td>Ethicon</td>
			<td>A305H</td>
			<td>Suture material for Langendorff apparatus</td>
		</tr>
		<tr>
			<td>Silk 5-0</td>
			<td>Ethicon</td>
			<td>A302H</td>
			<td>Suture material for carotid</td>
		</tr>
		<tr>
			<td>Small animal anesthesia workstation</td>
			<td>Hallowell EMC</td>
			<td>000A2770</td>
			<td>Small animal ventilator</td>
		</tr>
		<tr>
			<td>Sodium bicarbonate</td>
			<td>Milipore-Sigma</td>
			<td>S5761</td>
			<td>NaHCO3 &#8805;99,5% Part of the Krebs solution</td>
		</tr>
		<tr>
			<td>Sodium Chloride</td>
			<td>Milipore-Sigma</td>
			<td>S7653</td>
			<td>NaCl &#8805;99.5% Part of the Krebs solution</td>
		</tr>
		<tr>
			<td>Sodium Hydroxide pellets</td>
			<td>ACP chemicals</td>
			<td>S3700</td>
			<td>Diluted to 5N (10 g in 50 ml) for pH correction</td>
		</tr>
		<tr>
			<td>Sodium phosphate monobasic</td>
			<td>Milipore-Sigma</td>
			<td>S0751</td>
			<td>NaH2PO4 &#8805;99.0% Part of the Krebs solution</td>
		</tr>
		<tr>
			<td>Stevens Tenotomy Sciss, Str, Delicate, SH/SH, 4 1/2&#34;</td>
			<td>Skalar</td>
			<td>22-1240</td>
			<td>Small scisors for atria and cava vein opening</td>
		</tr>
		<tr>
			<td>Tissue slicer blades</td>
			<td>Thomas scientific</td>
			<td>6727C18</td>
			<td>Straight carbon steel blades for tissue slicing at the end of the protocol</td>
		</tr>
		<tr>
			<td>Tuberculin safety syringe with needle 25G 5/8&#34;</td>
			<td>CardinalHealth</td>
			<td>8881511235</td>
			<td>For heparin injection</td>
		</tr>
		<tr>
			<td>Veterinary General Surgery Set</td>
			<td>Skalar</td>
			<td>98-1275</td>
			<td>Surgery instruments including disection scisors and mosquito clamps</td>
		</tr>
		<tr>
			<td>Veterinary Micro Set</td>
			<td>Skalar</td>
			<td>98-1311</td>
			<td>Surgery instruments with microscisors used for carotid artery opening</td>
		</tr>
		<tr>
			<td>Working Heart Rat/Guinea Pig/Rabbit system</td>
			<td>Radnoti</td>
			<td>120101BEZ</td>
			<td>Modular working heart system modified for the needs of the protocol. Includes all the necesary tubbing, water jacketed reservoirs and valves, including 2 and 3 way stop cock</td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Bayer</td>
			<td></td>
			<td>Sedative. 20 mg/ml</td>
		</tr>
	</tbody>
</table>

pre-clinical model of cardiac donation after circulatory death

La demande de transplantation cardiaque est à la hausse; néanmoins, la disponibilité des organes est limitée en raison du manque de donneurs appropriés. Le don d'organes après la mort circulatoire (DCD) est une solution pour répondre à cette disponibilité limitée, mais en raison d'une période d'ischémie chaude prolongée et le risque de lésions tissulaires, son utilisation courante dans la transplantation cardiaque est rarement vu. Dans ce manuscrit, nous fournissons un protocole détaillé imitant étroitement les pratiques cliniques actuelles dans le contexte de DCD avec la surveillance continue de la fonction cardiaque, permettant l'évaluation des nouvelles stratégies et interventions cardioprotectrices pour diminuer ischémie-reperfusion.
Dans ce modèle, le protocole DCD est initié chez les rats Lewis anesthésiés en arrêtant la ventilation pour induire la mort circulatoire. Lorsque la pression artérielle systolique descend en dessous de 30 mmHg, le temps ischémique chaud est lancé. Après une période ischémique chaude pré-fixée, les coeurs sont rincés avec une solution cardioplégique normothermic, acheté, et monté sur un système de perfusion de coeur ex vivo de Langendorff. Après 10 min de réperfusion et de stabilisation initiales, le reconditionnement cardiaque est continuellement évalué pendant 60 min en utilisant la surveillance intraventriculaire de pression. Une lésion cardiaque est évaluée en mesurant la troponine cardiaque T et la taille infarctus est quantifiée par la coloration histologique. Le temps ischémique chaud peut être modulé et adapté pour développer la quantité désirée de dommages structurels et fonctionnels. Ce protocole simple permet l'évaluation de différentes stratégies de conditionnement cardioprotective introduites au moment de la cardioplégie, de la réperfusion initiale et/ou pendant la perfusion ex vivo. Les résultats obtenus à partir de ce protocole peuvent être reproduits dans de grands modèles, facilitant la traduction clinique.

Après l'extubation, la pression artérielle diminue rapidement dans un modèle prévisible (Figure 3). Le temps de mort prévu est inférieur à 5 min.
La figure 4 montre une courbe moyenne de pression/temps au début du reconditionnement après 0, 10 et 15 min de WIT. La fonction contractile s'améliorera avec le temps. L'utilisation de courtes périodes de SRT permettra à la passation de pouvoir de revenir à la normale, et les do...

Watch this Scientific Journal Video about Pre-clinical Model of Cardiac Donation after Circulatory Death at JoVE.com

Pre-clinical Model of Cardiac Donation after Circulatory Death

Ce protocole montre une approche simple et flexible pour l'évaluation de nouveaux agents de conditionnement ou de stratégies pour augmenter la faisabilité du don cardiaque après la mort circulatoire.

Modèle préclinique du don cardiaque après la mort circulatoire

Cardiac transplantation demand is on the rise; nevertheless, organ availability is limited due to a paucity of suitable donors. Organ donation after ...

Ce protocole cible la question limitée de l’approvisionnement en organes de transplantation en facilitant l’évaluation des différentes stratégies de conditionnement cardioprotecteur pour permettre l’utilisation de cœurs donnés après la mort circulatoire. Placer l’animal sur un coussin chauffant réglé à moyen. Après avoir confirmé un manque de réponse au pincement des pieds chez un rat adulte anesthésié, insérez une sonde rectale de température.Fixez un capteur transdermique d’oxymètre d’impulsion à une patte arrière, et couvrez le rat avec un tampon absorbant pour maintenir la température corporelle. Faire une incision de trois à quatre centimètres de la peau midline dans le cou, et utiliser des ciseaux incurvés pointe émoussée pour émousser disséquer le tissu sous-cutané, pour exposer le muscle sternohyoid droit. À l’aide de forceps non traumatiques, déplacez le muscle latéralement jusqu’à ce que l’artère carotide droite, la veine jugulaire et le nerf vague soient identifiés.Utilisez des ciseaux pour séparer soigneusement le nerf vague de l’artère carotide. Injecter 2000 unités internationales par kilogramme d’héparine dans la veine jugulaire droite, en appliquant une pression sur le site d’injection après la rétractation de l’aiguille pour éviter les fuites de sang. Utilisez des forceps incurvés pour passer deux sutures de soie 5-0 autour de l’artère carotide.Attachez fermement la suture distale pour occluser l’artère carotide à l’aspect supérieur de l’artère exposée, en gardant la suture proximale déliée, et placez le rat sous un microscope stéréo. Utilisez ensuite des ciseaux de microchirurgie pour faire soigneusement une incision d’un millimètre au-dessus de la paroi antérieure de l’artère carotide, et insérez un cathéter intraveineux fermé de 22 calibres et d’un pouce vers l’arc aortique. L’utilisation d’outils de grossissement et de microdissection est fortement recommandé pour ces étapes.Un champ propre peut être réalisé en tensionnant la suture proximale pour éviter le saignement. Le don cardiaque est nécessaire pour assembler l’appareil ex vivo cardioplégie. Pour initier le protocole DCD, d’abord extuber l’animal, puis utiliser des pinces à moustiques pour serrer la trachée, en commençant la phase agonale.Commencez à compter l’ESPRIT fonctionnel lorsque la pression artérielle systolique maximale descend en dessous de 30 millimètres de mercure ou d’asystole ou de fibrillation ventriculaire apparaît, selon la première. À la fin de WIT, effectuer une sternotomie médiale, en gardant le thorax ouvert avec un rétracteur Alm si nécessaire. Séparez le cœur des veines pulmonaires et d’autres structures thoraciques pour compléter la cardiectomie.Submergez immédiatement le cœur dans la solution krebs glacée pour un transport rapide vers le système ex vivo. Utilisez une suture en soie 2-0 pour fixer étroitement l’aorte à l’appareil Langendorff, puis ouvrez le robinet d’arrêt. Maintenant, ouvrez complètement le flux à la canule, et commencer le temps de reperfusion initiale et de stabilisation.Faites pivoter le cœur de sorte que l’atria est face au capteur de pression, et disséquer les veines pulmonaires pour élargir l’ouverture ventriculaire gauche auriculaire si nécessaire. Ensuite, insérez un ballon en latex relié à un capteur de pression, en vous assurant que le ballon est entièrement positionné à l’intérieur du ventricule. Remplissez lentement le ballon de solution saline jusqu’à ce que la pression diastolique finale atteigne 15 millimètres de mercure.Insérez l’électrode arpentante dans la face antérieure du cœur sans percer les vaisseaux coronaires. Une fois que les battements spontanés sont observés, réglez le rythme à 300 battements par minute. Après 10 minutes de stabilisation, lancez l’enregistrement continu de mesure de la pression intraventriculaire pour commencer la phase de reconditionnement et d’évaluation.Après une heure, retirer le cœur de l’appareil Langendorff et utiliser une lame droite en acier à haute teneur en carbone pour enlever les airs. Avec le ventricule droit orienté vers le bas, couper des tranches ventriculaires transversales d’un à deux millimètres d’épaisseur. Exciser le ventricule droit de la troisième tranche de tissu, et casser geler le ventricule gauche pour les analyses biochimiques en aval.Submergez les sections restantes dans la solution 5% TTC fraîchement préparée dans PBS pendant 10 minutes à 37 degrés Celsius. Le lendemain matin, lavez les échantillons deux fois dans du PBS frais et submergez les échantillons dans du PBS frais pour l’imagerie. Les tissus viables apparaîtront de couleur rouge brique.Après l’extubation, la pression artérielle diminue rapidement dans un modèle prévisible et le temps prévu à la mort est inférieur à cinq minutes. Ici, la pression moyenne par rapport aux courbes de temps au début du reconditionnement après zéro, 10 et 15 minutes de WIT sont affichées. La fonction contractile s’améliorera au fil du temps, l’utilisation de courtes périodes de WIT permettra à la contractilité de revenir à la normale, et les dommages morphologiques ne seront pas détectables.L’utilisation d’un agent de conditionnement ajouté avec la cardioplégie et à la phase de stabilisation a montré que les dommages générés par 15 minutes de WIT dans ce modèle sont agréables à la modulation par des agents cardioprotecteurs. Cette technique permet un contrôle total de toutes les variables pertinentes et des tests pharmacologiques et non pharmacologiques d’intervention et peut être reproduite dans des modèles plus grands, facilitant ainsi la traduction clinique.

pre-clinical-model-of-cardiac-donation-after-circulatory-death

Research

JoVE Journal

Medicine

Pre-clinical Evaluation of Tyrosine Kinase Inhibitors for Treatment of Acute Leukemia

Receptor tyrosine kinases have been implicated in the development and progression of many cancers, including both leukemia and solid tumors, and are attractive druggable therapeutic targets. Here we describe an efficient four-step strategy for pre-clinical evaluation of tyrosine kinase inhibitors (TKIs) in the treatment of acute leukemia. Initially, western blot analysis is used to confirm target inhibition in cultured leukemia cells. Functional activity is then evaluated using clonogenic assays in methylcellulose or soft agar cultures. Experimental compounds that demonstrate activity in cell culture assays are evaluated in vivo using NOD-SCID-gamma (NSG) mice transplanted orthotopically with human leukemia cell lines. Initial in vivo pharmacodynamic studies evaluate target inhibition in leukemic blasts isolated from the bone marrow. This approach is used to determine the dose and schedule of administration required for effective target inhibition. Subsequent studies evaluate the efficacy of the TKIs in vivo using luciferase expressing leukemia cells, thereby allowing for non-invasive bioluminescent monitoring of leukemia burden and assessment of therapeutic response using an in vivo bioluminescence imaging system. This strategy has been effective for evaluation of TKIs in vitro and in vivo and can be applied for identification of molecularly-targeted agents with therapeutic potential or for direct comparison and prioritization of multiple compounds.

Receptor tyrosine kinases are ectopically expressed in many cancers and have been identified as therapeutic targets in acute leukemia. This manuscript describes an efficient strategy for pre-clinical evaluation of tyrosine kinase inhibitors for the treatment of acute leukemia.

Évaluation pré-clinique des inhibiteurs de tyrosine kinase pour le traitement de la leucémie aiguë

Receptor tyrosine kinases have been implicated in the development and progression of many cancers, including both leukemia and solid tumors, and are ...

Recherche

Médecine

Handling of the Cotton Rat in Studies for the Pre-clinical Evaluation of Oncolytic Viruses

Oncolytic viruses are a novel anticancer therapy with the ability to target tumor cells, while leaving healthy cells intact. For this strategy to be successful, recent studies have shown that involvement of the host immune system is essential. Therefore, oncolytic virotherapy should be evaluated within the context of an immunocompetent model. Furthermore, the study of antitumor therapies in tolerized animal models may better recapitulate results seen in clinical trials. Cotton rats, commonly used to study respiratory viruses, are an attractive model to study oncolytic virotherapy as syngeneic models of mammary carcinoma and osteosarcoma are well established. However, there is a lack of published information on the proper handling procedure for these highly excitable rodents. The handling and capture approach outlined minimizes animal stress to facilitate experimentation. This technique hinges upon the ability of the researcher to keep calm during handling and perform procedures in a timely fashion. Finally, we describe how to prepare cotton rat mammary tumor cells for consistent subcutaneous tumor formation, and how to perform intratumoral and intraperitoneal injections. These methods can be applied to a wide range of studies furthering the development of the cotton rat as a relevant pre-clinical model to study antitumor therapy.

Cotton rats are extremely excitable and have a strong flight-or-fight response. A handling method optimized to reduce the stress of the animals is described which will make cotton rats more accessible as a preclinical model.

Manipulation du Rat coton en études pour l&#39;évaluation pré-clinique de virus oncolytiques

Oncolytic viruses are a novel anticancer therapy with the ability to target tumor cells, while leaving healthy cells intact.  For this strategy to be ...

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information about this disease. The human orthotopic prostate cancer xenograft mouse model provides a useful alternative approach for understanding the specific interactions between genetically and molecularly altered tumor cells, their organ microenvironment, and for evaluation of efficacy of therapeutic regimens. This is a well characterized model designed to study the molecular events of primary tumor development and it recapitulates the early events in the metastatic cascade prior to embolism and entry of tumor cells into the circulation. Thus it allows elucidation of molecular mechanisms underlying the initial phase of metastatic disease. In addition, this model can annotate drug targets of clinical relevance and is a valuable tool to study prostate cancer progression. In this manuscript we describe a detailed procedure to establish a human orthotopic prostate cancer xenograft mouse model.

Prostate cancer is the second most common cause of cancer-related deaths in the United States. An orthotopic cancer model provides a useful approach to understand the biology of prostate cancer and to evaluate the efficacy of therapeutic regimens. This protocol describes detailed steps necessary to establish an orthotopic prostate cancer mouse model.

Pré-clinique Orthotopique modèle murin de cancer de la prostate humaine

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

Use of a Low-flow Digital Anesthesia System for Mice and Rats

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers utilize a syringe pump to directly administer volatile anesthetics into a gas stream. Unlike a traditional vaporizer, it can be used at very low flow rates, making it ideal for use on mice and rats. 
The equipment's capability to use low flow rates could result in a substantial cost savings due to the reduced need for anesthetic agents, compressed gas, and charcoal scavenging filters1. A lower flow rate means less waste of anesthetic gas and likely reduces the risk of anesthetic exposure to laboratory personnel. Thus, the high levels of precision and safety associated with direct injection vaporizers, along with a reduced need for anesthetic agents, compressed gas, and charcoal filters are beneficial for research requiring small animal anesthesia. 
The goal of this protocol is to demonstrate the use of a syringe-driven direct injection vaporizer as part of a digital, low-flow anesthesia system. The direct injection vaporizer is capable of accurately delivering anesthesia at very low flow rates compared to a traditional vaporizer, making it a promising alternative for controlled gas anesthetic delivery to rodents.

Here, we present a protocol to more safely and efficiently administer anesthetic gas to mice using a digital, low flow anesthesia system utilizing a syringe-driven direct injection vaporizer.

Utilisation d&#39;un système d&#39;anesthésie numérique à faible débit pour les souris et les rats

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Mesure de la contractilité isométrique de l’artère mésentérique de souris à l’aide de fil myographie

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Implantation of hiPSC-derived Cardiac-muscle Patches after Myocardial Injury in a Guinea Pig Model

Due to the limited regeneration capacity of the heart in adult mammals, myocardial infarction results in an irreversible loss of cardiomyocytes. This loss of relevant amounts of heart muscle mass can lead to the heart failure. Besides heart transplantation, there is no curative treatment option for the end-stage heart failure. In times of organ donor shortage, organ independent treatment modalities are needed. Left-ventricular assist devices are a promising therapy option, however, especially as destination therapy, limited by its side-effects like stroke, infections and bleedings. In recent years, several cardiac repair strategies including stem cell injection, cardiac progenitors or myocardial tissue engineering have been investigated. Recent improvements in cell biology allow for the differentiation of large amounts of cardiomyocytes derived from human induced pluripotent stem cells (iPSC). One of the cardiac repair strategies currently under evaluation is to transplant artificial heart tissue. Engineered heart tissue (EHT) is a three-dimensional in vitro created cardiomyocyte network, with functional properties of native heart tissue. We have created EHT-patches from hiPSC derived cardiomyocytes. Here we present a protocol for the induction of left ventricular myocardial cryoinjury in a guinea pig, followed by implantation of hiPSC derived EHT on the left ventricular wall.

Here we present a protocol for the induction of left ventricular cryoinjury followed by the implantation of a cardiac muscle patch, derived from human iPS-cell cardiomyocytes in a guinea pig model.

Implantation des dérivés hiPSC myocarde patchs après lésion myocardique dans un modèle de cochon d’Inde

Due to the limited regeneration capacity of the heart in adult mammals, myocardial infarction results in an irreversible loss of cardiomyocytes. This loss ...

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure. Multiple small-animal models of heart transplantation have been used to study the acute and long-term effects of novel therapies. However, few are tested and demonstrated success in clinical trials. It is of critical importance to evaluate new therapies in a clinically relevant large-animal model for efficient and reliable translation of basic studies&#39;&#160;findings. Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and previously used to investigate novel cardioprotective strategies. This procedure focuses on acute ischemia-reperfusion injury and is a reliable method to investigate novel interventions which have been tested and validated in smaller experimental models, such as the murine model. We demonstrate its usefulness in assessing cardiac performance during the early post-transplantation period and other potential possibilities enabled by the model.

Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and utilized to investigate novel cardioprotective strategies.

Un modèle porcin préclinique de transplantation orthotopic de coeur

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart ...

Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. However, the evaluation of right ventricular pressure (RVP) and pulmonary artery pressure (PAP) remains technically challenging in mice. RVP and PAP are more difficult to measure than left ventricular pressure because of the anatomical differences between the left and right heart systems. In this paper, we describe a stable right heart hemodynamic measurement method and its validation using healthy and PAH mice. This method is based on open-chest surgery and mechanical ventilation support. It is a complicated procedure compared to closed chest procedures. While a well-trained surgeon is required for this surgery, the advantage of this procedure is that it can generate both RVP and PAP parameters at the same time, so it is a preferable procedure for the evaluation of PAH models.

Here, we present a protocol to perform an invasive hemodynamic assessment of the right ventricle and pulmonary artery in mice using an open-chest surgery approach.

Évaluation hémodynamique invasive pour le système ventriculaire droit et l'hypertension artérielle pulmonaire induite par l'hypoxie chez les souris

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many areas of biomedical research due to its cost-effectiveness and its translation to humans. This protocol describes a method to create a preclinical free skin flap model in rats with ischemia-reperfusion injury. The described 3 cm x 6 cm rat free skin flap model is easily obtained after the placement of several vascular ligatures and the section of the vascular pedicle. Then, 8 h after the ischemic insult and completion of the microsurgical anastomosis, the free skin flap develops the tissue damage. These ischemia-reperfusion injury-related damages can be studied in this model, making it a suitable model for evaluating therapeutic agents to address this pathophysiological process. Furthermore, two main monitoring techniques are described in the protocol for the assessment of this animal model: transit-time ultrasound technology and laser speckle contrast analysis.

Here, we describe a pre-clinical animal model for studying the pathophysiology of ischemia-reperfusion injury in reconstructive microsurgery. This free skin flap model based on the superficial caudal epigastric vessels in the rat may also allow for the evaluation of different therapies and compounds to counteract ischemia-reperfusion injury-related damage.

Un modèle préclinique de rat pour l'étude des dommages d'ischémie-réperfusion dans la microchirurgie reconstructive

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Use of an Integrated Low-Flow Anesthetic Vaporizer, Ventilator, and Physiological Monitoring System for Rodents

Low-flow digital vaporizers commonly utilize a syringe pump to directly administer volatile anesthetics into a stream of carrier gas. Per animal welfare recommendations, animals are warmed and monitored during procedures requiring anesthesia. Common anesthesia and physiological monitoring equipment include gas tanks, anesthetic vaporizers and stands, warming controllers and pads, mechanical ventilators, and pulse oximeters. A computer is also necessary for data collection and to run equipment software. In smaller spaces or when performing field work, it can be challenging to configure all this equipment in limited space.
The goal of this protocol is to demonstrate best practices for use of a low-flow digital vaporizer using both compressed oxygen and room air, along with an integrated mechanical ventilator, pulse oximeter, and far infrared warming as an all-inclusive anesthesia and physiological monitoring suite ideal for rodents.

Here, we present a protocol to safely and effectively administer anesthetic gas to mice using a digital, low flow anesthesia system with integrated ventilator and physiological monitoring modules.

Utilisation d’un vaporisateur anesthésique intégré à faible débit, d’un ventilateur et d’un système de surveillance physiologique pour les rongeurs

Low-flow digital vaporizers commonly utilize a syringe pump to directly administer volatile anesthetics into a stream of carrier gas. Per animal welfare ...