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

Bu çalışmanın bazı bölümleri, Fondation Marcel et Rolande Gosselin ve Fondation Bay stefane Foumy tarafından cömert bir katkı tarafından destekleniyor. Nicolas Noiseux FRQ-S bilim adamı.
Yazarlar veri toplama destek için Josh Zhuo Le Huang, Gabrielle Gascon, Sophia Ghiassi ve Catherine Scalabrini teşekkür etmek istiyorum.

Laboratuar hayvanlarının bakımı ve kullanımı kılavuzu 'na uygun tüm hayvan bakımı ve deneysel protokoller, Centre Hospitalier de l 'Université de Montréal araştırma merkezi 'nin kurumsal hayvan bakımı ve kullanım Komitesi tarafından onaylanmıştır.
1. ön preparatlar
<ol><li>Kardiyoplegia teslimat sistemini (Şekil 1a) ve Langendorff ex vivo perfüzyon sistemini (Şekil 1B) ısıtmak için su banyosunu açın. 37...

<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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Burada sunulan protokol kalp DCD basit, rahat ve çok yönlü bir model tanıttı, kalp fonksiyonel kurtarma değerlendirmek için fırsat sunan, doku hasarı ve donör kurtarma geliştirmek için sonrası Klima kardiyokoruyucu ajanlar kullanımı kalp nakli için aksi atılır. Ex vivo kalp perfüzyon sistemleri (EVHP) sistemleri, kardiyak fonksiyonun değerlendirilmesi için bir platform sağlamak üzere optimize edilmiştir ve post-Conditioning farmakolojik ajanlar ile tamamlayıcı olarak değiştirilmiş çözümler...

Genel ve kalp transplantasyonunda katı organ nakli özellikle dünya çapında1,2' de yükselir. Organ alımı standart yöntemi beyin ölümünden sonra bağış (DBD). DBD katı dahil kriterleri göz önüne alındığında, sunulan kalplerin% 40 daha az kabul edilir3, böylece artan talep karşısında teklif sınırlandırmak ve organ bekleme listesini uzatmak. Bu sorunu gidermek için, dolaşım ölümü (DCD) sonrasında bağışlanan organların kullanımı potansiyel bir çözüm olarak kabul edilir4.
DCD bağış, ancak, bakım ve resüsitasyon önce korunmasız sıcak iskemi bir süre geri çekilmesi aşağıdaki bir agonal faz kaçınılmaz5. Dolaşım ölümünden sonra potansiyel organ hasarı organ fonksiyon bozukluğu neden olabilir, rutin DCD kalp nakli benimsemeye isteksizlik açıklayan. Bu sadece 4 merkezleri klinik olarak, çok kısa sıcak iskemi kez ve kronik patolojiler olmadan genç bağış içeren sıkı kriterler ile DCD Hearts kullanmak bildirilmiştir5/6. Etik ve yasal nedenlerden dolayı, dolaşım ölümü5,8,9öncesinde bağışçılardan sınırlı veya hiçbir kardiyokoruyucu müdahaleler uygulanabilir. Böylece, iskemi-reperfüzyon (IR) yaralanmasını hafifletmek için herhangi bir azaltma kardiyoplegik çözümler ile erken reperfüzyon sırasında başlatılan kardiyokoruyucu tedaviler ile sınırlıdır ve uygun fonksiyonel değerlendirme için izin vermez. Ex vivo kalp perfüzyon (evhp) ve özel platformlar kullanarak DCD kalp yenileme alternatif bir çözüm olarak önerilen ve çeşitli akademisyenler tarafından okudu10,11,12,13 . EVHP fonksiyonel kurtarma geliştirmek için DCD kalpleri sonrası Klima ajanlar sunmak için benzersiz bir fırsat sunuyor. Ancak, verimli klinik çeviri için, birçok teknik ve pratik sorunlar ele alınması için, ve bu daha fazla perfüzyon ve fonksiyonel kriterler bir dizi üzerinde uzlaşma eksikliği tarafından transplantability belirlemek için bileşik6, 8' den itibaren.
Burada, ilk reperfüzyon sırasında, satın alma sırasında başlatılan organ sonrası Klima araştırması için kullanılabilecek bir ex vivo kalp perfüzyon sistemi ile birlikte tekrarlanabilir öncesi klinik küçük hayvan DCD protokolünün gelişmesini bildiriyoruz ve /veya EVHP boyunca.

<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

Kardiyak transplantasyon talebi yükselişte; Yine de, organ mevcudiyeti uygun bağış nedeniyle sınırlıdır. Dolaşım ölümünden sonra organ bağış (DCD) bu sınırlı kullanılabilirliği gidermek için bir çözümdür, ancak uzun süreli sıcak iskemi ve doku yaralanması riski nedeniyle, kardiyak transplantasyon rutin kullanımı nadiren görülür. Bu yazıda, kalp fonksiyonunun sürekli izlenmesi ile DCD bağlamında mevcut klinik uygulamaları yakından taklit eden ayrıntılı bir protokol sağlıyoruz, yeni kardiyokoruyucu stratejilerin değerlendirilmesine ve müdahalelerin azalmasına izin verme iskemi-reperfüzyon hasarı.
Bu modelde, DCD protokol dolaşım ölümü teşvik etmek için havalandırma durdurularak anestezi Lewis fareler başlatılır. Sistolik kan basıncı 30 mmHg 'ye düştüğünde sıcak iskemik zaman başlatılır. Önceden ayarlanan sıcak iskemik döneminden sonra, kalpler bir normothermic kardiyoplegik solüsyon ile boşaltılır, temin edilir ve bir Langendorff ex vivo kalp perfüzyon sistemine monte edilir. İlk reperfüzyon ve stabilizasyonu 10 dk sonra, kalp yenileme intraventriküler basınç izleme kullanarak 60 dk için sürekli olarak değerlendirilir. Kalp hasarı, kardiyak troponin T ölçümüne göre değerlendirilir ve infarkıt büyüklüğü histolojik lekelenme ile ölçülebilir. Sıcak iskemik zaman modülasyonlu ve yapısal ve fonksiyonel hasar istenen miktarda geliştirmek için uyarlanmış olabilir. Bu basit protokol kardiyoplegia, ilk reperfüzyon ve/veya ex vivo perfüzyon sırasında sunulan farklı kardiyokoruyucu Klima stratejilerinin değerlendirilmesi için izin verir. Bu protokolden elde edilen bulgular büyük modellerde yeniden üretilerek klinik çeviriyi kolaylaştırabilir.

Uzama sonrasında kan basıncı öngörülebilir bir desenle hızla düşer (Şekil 3). Beklenen ölüm süresi 5 dakikadan az.
Şekil 4 , 0, 10 ve 15 dk. WIN sonrasında yeniden yenileme başlangıcında ortalama bir basınç/zaman eğrisi gösterir. Kontril fonksiyonu zamanla gelişecektir. WıN kısa sürelerinin kullanımı kontrazlığın normale dönmesini sağlar ve morfolojik hasar algılanamaz (Şekil 5</...

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

Bu protokol, yeni Klima ajanları veya dolaşım ölümünden sonra kardiyak bağış fizibilitesi artırmak için stratejiler değerlendirilmesi için basit ve esnek bir yaklaşım gösterir.

Dolaşım ölümünden sonra kardiyak bağış öncesi klinik model

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

Bu protokol, dolaşım ölümünden sonra bağışlanan kalplerin kullanımına olanak sağlayacak farklı kardiyoprotektif kondisyon stratejilerinin değerlendirilmesini kolaylaştırarak sınırlı organ nakli organ temini konusunu hedeflemektedir. Orta ayarlanmış bir ısıtma yastığı üzerinde hayvan yerleştirin. Anesteziedilmiş yetişkin sıçanda parmak ucuna yanıt eksikliğini doğruladıktan sonra rektal sıcaklık sondası yerleştirin.Bir arka pençe bir transdermal darbe oksimetre sensörü takın ve vücut ısısını korumak için bir emici ped ile sıçan kapağı. Boyunda üç-dört santimetre orta hat deri kesi yapmak, ve künt ucu kavisli makas kullanın deri altı doku künt diseksiyon, sağ sternohyoid kas ortaya çıkarmak için. Nontravmatik forceps kullanarak, sağ karotis arter kadar yanal kas hareket, juguler ven, ve vagus siniri.Vagus sinirini karotis arterden dikkatlice ayırmak için makas kullanın. Sağ juguler vene kilogram başına 2,000 uluslararası ünite enjekte edin, kan sızıntısını önlemek için iğne geri çektikten sonra enjeksiyon bölgesine basınç uygulayın. Karotis arter etrafında iki 5-0 ipek dikiş geçmek için kavisli forceps kullanın.Sıkıca maruz arterin üstün yönü karotis arter tıkanıklığı için distal sütür takın, proksimal dikiş çözülmüş tutmak, ve bir stereo mikroskop altında sıçan yerleştirin. Sonra dikkatle karotis arterin ön duvarı üzerinde bir milimetrelik kesi yapmak için mikrocerrahi makas kullanın, ve aort kemer doğru 22-gauge, bir inç kapalı intravenöz kateter eklemek. Büyütme ve mikrodisese araçlarının kullanımı bu adımlar için son derece tavsiye edilir.Kanamayı önlemek için proksimal sütür gerilerek temiz bir alan elde edilebilir. Kardiyak bağış ex vivo cardioplegia cihazı monte etmek için gereklidir. DCD protokolünü başlatmak için önce hayvanı ektübasyona, sonra da nefes borusunu kenetlemek için sivrisinek forcep'leri kullanarak agonal evreye başla.En yüksek sistolik kan basıncı 30 milimetrenin altına düştüğünde fonksiyonel WIT'yi saymaya başlayın. WIT sonunda, gerekirse bir Alm retraktör ile toraks açık tutarak, bir medial sternotomi gerçekleştirin. Kardiyoektomi tamamlamak için pulmoner venler ve diğer torasik yapılardan kalbi ayırın.Ex vivo sistemine hızlı bir ulaşım için kalbi hemen buz gibi Krebs çözümüne daldırın. Langendorff cihazına aortu sıkıca tamir etmek için 2-0 ipek sütür kullanın ve sonra stopcock açın. Şimdi kanüle akışı tamamen açın ve ilk reperfüzyon ve stabilizasyon süresini başlatın.Kalbi, atria basınç sensörüne bakacak şekilde döndürün ve gerekirse sol ventrikül atriyal açıklığı genişletmek için pulmoner damarları inceleyin. Daha sonra, bir basınç sensörüne bağlı bir lateks balon takın ve balonun ventrikülün içine tam olarak yerleştirildiğinden emin olun. Yavaşça son diyastolik basınç cıva 15 milimetre ulaşana kadar tuzlu balon doldurun.Koroner damarları delmeden kalbin ön yüzüne pacing elektrodu yerleştirin. Spontan dayak gözlendikten sonra, dakikada 300 atım için pacing ayarlayın. 10 dakikalık stabilizasyondan sonra, yenileme ve değerlendirme aşamasına başlamak için sürekli intraventriküler basınç ölçüm kaydını başlatın.Bir saat sonra, Langendorff aparatından kalbi çıkarın ve atria'yı çıkarmak için düz yüksek karbonlu çelik bıçak kullanın. Sağ ventrikül aşağı bakacak şekilde, bir ila iki milimetre kalınlığında enine ventriküler dilimler kesin. Doku üçüncü dilim sağ ventrikül çıkarmak ve snap downstream biyokimyasal analizler için sol ventrikül dondurmak.Kalan bölümleri PBS'de yeni hazırlanmış %5'lik TTC çözeltisine 37 derecede 10 dakika batırın. Ertesi sabah, numuneleri taze PBS'de iki kez yıkayın ve görüntüleme için numuneleri taze PBS'ye batırın. Canlı dokular tuğla-kırmızı renkli görünür.Ekstübasyondan sonra, kan basıncı hızla öngörülebilir bir model düşer ve ölüm beklenen zaman az beş dakikadır. Burada, sıfır, 10 ve 15 dakikalık WIT'yi takiben yenilemenin başlangıcındaki ortalama basınç ve zaman eğrileri gösterilir. Kontraktil fonksiyon zamanla düzelecek, WIT kısa sürekullanımı sözleşme normale dönmek için izin vemorf hasar tespit edilemez.Kardiyopleji ile eklenen bir kondisyon ajanının kullanımı ve stabilizasyon aşamasında bu modelde 15 dakikalık WIT'nin verdiği hasarın kardiyoprotektif ajanlar tarafından modülasyona uygun olduğunu göstermiştir. Bu teknik, ilgili tüm değişkenlerin ve farmakolojik ve farmakolojik olmayan müdahale testlerinin tam olarak kontrol altına alınmasına olanak sağlar ve daha büyük modellerde çoğaltılarak klinik çevirinin kolaylaştırılmasını sağlar.

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

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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.

Akut lösemi tedavisi için Tirosin Kinaz İnhibitörleri klinik öncesi değerlendirilmesi

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

Tıp

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.

Oncolytic virüs klinik öncesi Değerlendirme Çalışmaları Pamuk Rat Taşıma

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.

İnsan Prostat Kanseri klinik öncesi Ortotopik murin modeli

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.

Fare ve Sıçanlarda için düşük akış Dijital Anestezi Sistemi Kullanımı

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.

İzometrik Contractility ölçüm tel Myography kullanarak fare mezenterik arter

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.

HiPSC elde edilen kalp kası yamalar implantasyonu eskiden şiling şimdi domuz modelinde miyokardiyal yaralanma sonra

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.

Ortootopik kalp transplantasyonunun klinik öncesi porcine modeli

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.

Farelerde Sağ Ventrikül Sistemi ve Hipoksiye Bağlı Pulmoner Arteriyel Hipertansiyon için İnvaziv Hemodinamik Değerlendirme

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.

Rekonstrüktif Mikrocerrahide İskemi-Reperfüzyon Hasarı Nın İnceleştirilmesinde Klinik Öncesi Sıçan Modeli

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.

Kemirgenler için Entegre Düşük Akışlı Anestezik Buharlaştırıcı, Ventilatör ve Fizyolojik İzleme Sisteminin Kullanımı

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