Name: murine short axis ventricular heart slices for electrophysiological studies
Uploaded: 2017-06-04T14:00:00
Description: Watch this Scientific Journal Video about Murine Short Axis Ventricular Heart Slices for Electrophysiological Studies at JoVE.com

Vi anerkender den støtte, der ydes af workshoppen og dyreanlægget hos instituttet for neurofysiologi. Dette arbejde blev støttet af Walter und Marga Boll-Stiftung, Köln Fortune og Deutsche Stiftung für Herzforschung.

Dyrehåndtering skal være i overensstemmelse med retningslinjerne fra det lokale dyrevelfærdsudvalg og til Europa-Parlamentets direktiv 2010/63 / EU. 1. Forbered løsninger <ol><li> Forbered Tyrode&#39;s opløsning uden Ca 2+ (sammensætning i mM): NaCl 136, KCl 5.4, NaH2P03 0,33, MgCl2 1, glucose 10, HEPES 5, 2,3-butandionmonoxim (BDM) 30. Juster pH til 7,4 Med NaOH ved 4 ° C. </li><li> Klargør Tyrode&#39;s opløsning med Ca2 + (sammensætning i mM): NaCl 136, KC...

<ol>
	<li>Yamamoto, C., McIlwain, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+activities+in+thin+sections+from+the+mammalian+brain+maintained+in+chemically-defined+media+in+vitro.">Electrical activities in thin sections from the mammalian brain maintained in chemically-defined media in vitro.</a> J Neurochem. 13, 1333-1343 (1966).</li><li>Colbert, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+cortical+brain+slices+for+electrophysiological+recording.">Preparation of cortical brain slices for electrophysiological recording.</a> Methods Mol Biol. 337, 117-125 (2006).</li><li>Ad Graaf, I., Groothuis, G. M., Olinga, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Precision-cut+tissue+slices+as+a+tool+to+predict+metabolism+of+novel+drugs.">Precision-cut tissue slices as a tool to predict metabolism of novel drugs.</a> Expert Opin Drug Metab Toxicol. 3, 879-898 (2007).</li><li>Kim, Y. H., 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=Cardiopulmonary+toxicity+of+peat+wildfire+particulate+matter+and+the+predictive+utility+of+precision+cut+lung+slices.">Cardiopulmonary toxicity of peat wildfire particulate matter and the predictive utility of precision cut lung slices.</a> Part Fibre Toxicol. 11, 29 (2014).</li><li>Nembo, E. 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=In+vitro+chronotropic+effects+of+Erythrina+senegalensis+DC+(Fabaceae)+aqueous+extract+on+mouse+heart+slice+and+pluripotent+stem+cell-derived+cardiomyocytes.">In vitro chronotropic effects of Erythrina senegalensis DC (Fabaceae) aqueous extract on mouse heart slice and pluripotent stem cell-derived cardiomyocytes.</a> J Ethnopharmacol. 165, 163-172 (2015).</li><li>Wang, 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=Cardiac+tissue+slices:+preparation,+handling,+and+successful+optical+mapping.">Cardiac tissue slices: preparation, handling, and successful optical mapping.</a> Am J Physiol Heart Circ Physiol. 308, H1112-H1125 (2015).</li><li>Bussek, 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=Tissue+slices+from+adult+mammalian+hearts+as+a+model+for+pharmacological+drug+testing.">Tissue slices from adult mammalian hearts as a model for pharmacological drug testing.</a> Cell Physiol Biochem. 24, 527-536 (2009).</li><li>Burnashev, N. A., Edwards, F. A., Verkhratsky, A. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patch-clamp+recordings+on+rat+cardiac+muscle+slices.">Patch-clamp recordings on rat cardiac muscle slices.</a> Pflugers Arch. 417, 123-125 (1990).</li><li>Pillekamp, F., 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=Establishment+and+characterization+of+a+mouse+embryonic+heart+slice+preparation.">Establishment and characterization of a mouse embryonic heart slice preparation.</a> Cell Physiol Biochem. 16, 127-132 (2005).</li><li>Pillekamp, F., 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=Neonatal+murine+heart+slices.+A+robust+model+to+study+ventricular+isometric+contractions.">Neonatal murine heart slices. A robust model to study ventricular isometric contractions.</a> Cell Physiol Biochem. 20, 837-846 (2007).</li><li>Halbach, 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=Ventricular+slices+of+adult+mouse+hearts--a+new+multicellular+in+vitro+model+for+electrophysiological+studies.">Ventricular slices of adult mouse hearts--a new multicellular in vitro model for electrophysiological studies.</a> Cell Physiol Biochem. 18, 1-8 (2006).</li><li>Halbach, 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=Electrophysiological+maturation+and+integration+of+murine+fetal+cardiomyocytes+after+transplantation.">Electrophysiological maturation and integration of murine fetal cardiomyocytes after transplantation.</a> Circ. Res. 101, 484-492 (2007).</li><li>Halbach, 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=Time-course+of+the+electrophysiological+maturation+and+integration+of+transplanted+cardiomyocytes.">Time-course of the electrophysiological maturation and integration of transplanted cardiomyocytes.</a> J. Mol. Cell Cardiol. 53, 401-408 (2012).</li><li>Halbach, 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=Cell+persistence+and+electrical+integration+of+transplanted+fetal+cardiomyocytes+from+different+developmental+stages.">Cell persistence and electrical integration of transplanted fetal cardiomyocytes from different developmental stages.</a> Int. J. Cardiol. 171, e122-e124 (2014).</li><li>Halbach, 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=Electrophysiological+integration+and+action+potential+properties+of+transplanted+cardiomyocytes+derived+from+induced+pluripotent+stem+cells.">Electrophysiological integration and action potential properties of transplanted cardiomyocytes derived from induced pluripotent stem cells.</a> Cardiovasc. Res. 100, 432-440 (2013).</li><li>Verrecchia, F., Herve, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reversible+blockade+of+gap+junctional+communication+by+2,3-butanedione+monoxime+in+rat+cardiac+myocytes.">Reversible blockade of gap junctional communication by 2,3-butanedione monoxime in rat cardiac myocytes.</a> Am J Physiol. 272, C875-C885 (1997).</li><li>Watanabe, Y., 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=Inhibitory+effect+of+2,3-butanedione+monoxime+(BDM)+on+Na(+)/Ca(2+)+exchange+current+in+guinea-pig+cardiac+ventricular+myocytes.">Inhibitory effect of 2,3-butanedione monoxime (BDM) on Na(+)/Ca(2+) exchange current in guinea-pig cardiac ventricular myocytes.</a> Br J Pharmacol. 132, 1317-1325 (2001).</li><li>Fleischmann, B. 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=Differential+subunit+composition+of+the+G+protein-activated+inward-rectifier+potassium+channel+during+cardiac+development.">Differential subunit composition of the G protein-activated inward-rectifier potassium channel during cardiac development.</a> J Clin Invest. 114, 994-1001 (2004).</li><li>Peinkofer, G., 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=From+Early+Embryonic+to+Adult+Stage:+Comparative+Study+of+Action+Potentials+of+Native+and+Pluripotent+Stem+Cell-Derived+Cardiomyocytes.">From Early Embryonic to Adult Stage: Comparative Study of Action Potentials of Native and Pluripotent Stem Cell-Derived Cardiomyocytes.</a> Stem Cells Dev. 25, 1397-1406 (2016).</li></ol>

Ventrikulære skiver muliggør elektrofysiologiske, farmakologiske og mekaniske undersøgelser med en bevaret in vivo- lignende vævsstruktur og direkte adgang til målteknologien til alle områder i hjertet. Fysiologiske virkningspotentiale egenskaber er blevet påvist i embryonale, neonatale og voksne skiver 9 , 10 , 11 . Skiveens vitalitet, bortset fra overfladelagene, der er direkte beskadiget af skæreproceduren, ...

Tynde vævsskiver er blevet brugt ofte i grundforskningen, da Yamamot og Mcllwain i 1966 viste, at elektrisk aktivitet af hjerneskiver bevares in vitro 1 . Siden da er elektrofysiologiske og farmakologiske undersøgelser blevet udført på skiver fra hjerne 2 , lever 3 , lunge 4 og myokardvæv 5 , 6 , 7 . Første patch-clamp-optagelser i ventrikulære skiver fra neonatale rottehjerter blev beskrevet i 1990 8 , men denne teknik faldt i glemsel i nogen tid. Mere end et årti senere etablerede vores gruppe en ny metode til at forberede murine embryonale 9 , neonatal 10 og voksne 11 hjerte skiver. Disse levedygtige vævsskiver kan anvendes til akutte forsøg (voksen skiveS kan dyrkes i flere timer) eller kortsigtede kulturforsøg (embryonale og neonatale skiver kan dyrkes i nogle dage). Skiver viser in vivo som elektrofysiologiske karakteristika og en homogen excitationsudbredelse som vurderet ved skarpt elektrode-actionpotentiale og mikroelektrode-optagelser 11 . På grund af deres &quot;todimensionelle&quot; morfologi tillader de direkte adgang til optagelektroder til alle områder i ventriklen, hvilket gør dem til et interessant redskab til elektrofysiologiske undersøgelser og rejser nye forsøgsmuligheder i forhold til Langendorff-perfuserede hjerter. Lægemiddelrespons af skiverne til ionkanalblokkere som verapamil (L-type Ca 2+ -kanalblokker), lidokain (Na + -kanalblokker), 4-aminopyridin (ikke-spændingsafhængig K + -kanalblokker) og linopirdin (KCNQ K + -kanalblokker) 9 , 11 </suP&gt; svarede til kendte effekter på dissocierede kardiomyocytter. Isometriske kraftmålinger afslørede et positivt kraftfrekvensforhold og anbefalede stærkt intakt kontraktil funktion 10 . Disse resultater viste, at murine ventrikulære skiver er egnede som en in vitro vævsmodel til fysiologiske og farmakologiske undersøgelser. Desuden har ventrikulære skiver af modtagers hjerter i kombination med skarpe elektrodeoptagelser vist sig at være et meget nyttigt redskab til karakterisering af elektrisk og mekanisk integration samt modning af transplanterede føtal 12 , 13 , 14 og stamceller afledte 15 cardiomyocytter. Sammenfattende er ventrikulære skiver en værdifuld og veletableret multicellular vævsmodel og bør betragtes som komplementære til dissocierede cardiomyocytter og Langendorff-perfuserede hjerterI kardiovaskulær forskning med den største fordel ved at tilvejebringe en in vivo- lignende vævsstruktur (i modsætning til dissocierede celler) samt direkte adgang til måle teknologier som skarpe elektrodeoptagelser til alle områder i hjertet (i modsætning til hele hjertepræparater).

<table border="0" cellpadding="0" cellspacing="0" width="895">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:248px;">Leica VT 1000s</td>
			<td style="width:340px;">Leica Microsystems, Wetzlar, Germany</td>
			<td style="width:140px;"></td>
			<td style="width:167px;">Microtome with vibrating blade.</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:248px;">Stainless Steel Blades</td>
			<td style="width:340px;">Campden Instruments, Loughborough, England</td>
			<td>7550-1-SS</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pasteur pipettes&#160;</td>
			<td>Sigma-Aldrich, St. Louise, USA</td>
			<td>Z627992&#160;</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:248px;">Fine brush, e.g. size 6 (4/32&#34;)</td>
			<td style="width:340px;">VWR, International, Radnor, USA</td>
			<td>149-2125</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">Preparation table</td>
			<td style="width:340px;"></td>
			<td style="width:140px;"></td>
			<td style="width:167px;">self made</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:248px;">Molt for embedding ventricles in agarose</td>
			<td style="width:340px;"></td>
			<td style="width:140px;"></td>
			<td style="width:167px;">self made</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:248px;">1 ml Syringe</td>
			<td style="width:340px;">Becton, Dickinson;&#160; Franklin Lakes, USA</td>
			<td>300013</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">27Gx3/4`` Needles</td>
			<td rowspan="2" style="width:340px;">Braun, Melsungen, Germany</td>
			<td>4657705</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">20G 11/2`` Needles</td>
			<td>4657519</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">Small scissor</td>
			<td style="width:340px;">WPI, Sarasota, USA</td>
			<td>501263</td>
			<td rowspan="2" style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">Tweezers #5, 0.1 x 0.06 mm tip</td>
			<td style="width:340px;">WPI, Sarasota, USA</td>
			<td>500342</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">Oxygen gas (medical grade O2)</td>
			<td style="width:340px;">Linde, Munich, Germany</td>
			<td style="width:140px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">Carbogen gas (95 % O2, 5 % CO2)&#160;</td>
			<td style="width:340px;">Linde, Munich, Germany</td>
			<td style="width:140px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">NaCL</td>
			<td style="width:340px;">Sigma-Aldrich, St. Louise, USA</td>
			<td style="width:140px;">7647-14-5</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">KCL</td>
			<td style="width:340px;">Sigma-Aldrich, St. Louise, USA</td>
			<td style="width:140px;">746436</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:248px;">CaCl2</td>
			<td style="width:340px;">Sigma-Aldrich, St. Louise, USA</td>
			<td style="width:140px;">746495</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:248px;">KH2PO4</td>
			<td style="width:340px;">Sigma-Aldrich, St. Louise, USA</td>
			<td style="width:140px;">NIST200B&#160;</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">HEPES</td>
			<td style="width:340px;">Sigma-Aldrich, St. Louise, USA</td>
			<td style="width:140px;">51558</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:248px;">NaHCO3</td>
			<td style="width:340px;">Sigma-Aldrich, St. Louise, USA</td>
			<td style="width:140px;">S5761&#160;</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">D(+)-Glucose</td>
			<td style="width:340px;">Sigma-Aldrich, St. Louise, USA</td>
			<td style="width:140px;">G8270&#160;</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:248px;">MgSO4</td>
			<td style="width:340px;">Sigma-Aldrich, St. Louise, USA</td>
			<td style="width:140px;">M7506&#160;</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">NaOH</td>
			<td style="width:340px;">Sigma-Aldrich, St. Louise, USA</td>
			<td style="width:140px;">S8045&#160;</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">Cyanoacrylate glue&#160;</td>
			<td style="width:340px;">Henkel, D&#252;sseldorf, Germany</td>
			<td style="width:140px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:248px;">Low-melt Agarose&#160;</td>
			<td style="width:340px;">Roth, Karlsruhe, Germany</td>
			<td style="width:140px;">6351.2</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:248px;">Heparin-sodium-25000 I.E./5mL</td>
			<td style="width:340px;">Ratiopharm, Ulm, Germany</td>
			<td style="width:140px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:248px;">Dulbecco&#39;s Modified Eagle Medium&#160; (DMEM), high glucose, GlutaMAX</td>
			<td style="width:340px;">ThermoScientific, Waltham, USA</td>
			<td style="width:140px;">10566016</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:248px;">SEC-10LX Amplifier</td>
			<td style="width:340px;">npi electronic GmbH, Tamm, Germany</td>
			<td>SEC-10LX</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:248px;">EPC 9</td>
			<td style="width:340px;">HEKA Elektronik GmbH, Lambrecht, Germany</td>
			<td style="width:140px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:248px;">Zeiss Axiovert 200</td>
			<td style="width:340px;">Zeiss, Oberkochen, Germany</td>
			<td style="width:140px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:248px;">&#160;Low magnification Micromanipulator</td>
			<td style="width:340px;">Narashige, Tokyo, Japan</td>
			<td style="width:140px;">Nm-3</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:248px;">High magnification, three-axis micromanipulator</td>
			<td style="width:340px;">Narashige, Tokyo, Japan</td>
			<td style="width:140px;">MHW-3</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:248px;">Peristaltic perfusion pump</td>
			<td style="width:340px;">Multi Channel Systems, Reutlingen, Germany</td>
			<td style="width:140px;">PPS2</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">2-channel temperature controller</td>
			<td style="width:340px;">Multi Channel Systems, Reutlingen, Germany</td>
			<td style="width:140px;">TCO02</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:248px;">Square pulse stimulator</td>
			<td style="width:340px;">Natus Europe GmbH, Planegg, Germany</td>
			<td style="width:140px;">Grass SD9</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">Glass capillaries</td>
			<td style="width:340px;">WPI, Sarasota, USA</td>
			<td style="width:140px;">1B150F-1</td>
			<td style="width:167px;"></td>
		</tr>
	</tbody>
</table>

murine short axis ventricular heart slices for electrophysiological studies

Murine-kardiomyocytter er blevet anvendt i vid udstrækning til in vitro- undersøgelser af hjertefysiologi og nye terapeutiske strategier. Imidlertid er multicellulære præparater af dissocierede cardiomyocytter ikke repræsentative for den komplekse in vivo struktur af cardiomyocytter, ikke-myocytter og ekstracellulær matrix, som påvirker både mekaniske og elektrofysiologiske egenskaber i hjertet. Her beskriver vi en teknik til at forberede levedygtige ventrikulære skiver af voksne mushjerter med en bevaret in vivo- lignende vævsstruktur og demonstrere deres egnethed til elektrofysiologiske optagelser. Efter udskæring af hjertet separeres ventrikler fra atrierne, perfusioneres med Ca2 + -fri opløsning indeholdende 2,3-butandionmonoxim og indlejret i en 4% lavmeltig agaroseblok. Blokken er anbragt på en mikrotome med et vibrerende blad, og vævsskiver med en tykkelse på 150-400 μm er forberedt og holder vibrationen friBladets klinge ved 60-70 Hz og flytter bladet fremad så langsomt som muligt. Tykkelsen af ​​skiverne afhænger af den videre anvendelse. Skiver opbevares i iskold Tyrode&#39;s opløsning med 0,9 mM Ca 2+ og 2,3-butandionmonoxim (BDM) i 30 minutter. Derefter overføres skiver til 37 ° C DMEM i 30 minutter for at vaske ud BDM. Skiver kan anvendes til elektrofysiologiske undersøgelser med skarpe elektroder eller mikroelektroder, til kraftmålinger til analyse af kontraktil funktion eller for at undersøge interaktionen mellem transplanterede stamcellerafledte cardiomyocytter og værtsvæv. Til skarpe elektrodeoptagelser placeres en skive i en 3 cm cellekulturskål på varmepladen af ​​et inverteret mikroskop. Skiven stimuleres med en unipolær elektrode, og intracellulære aktionspotentialer af cardiomyocytter inden for skiven registreres med en skarp glaselektrode.

Myokardieinfarkt fører til et næsten uopretteligt tab af kardiomyocytter. Celleudskiftningsterapi ved anvendelse af stamcellerafledte cardiomyocytter til eksogen hjerteregenerering er en lovende terapeutisk tilgang. Elektrisk integration og modning af de transplanterede celler er afgørende for sikkerhed og effektivitet af celleudskiftningsterapi. For at vurdere integration og modning, transplanterede vi kardiomyocytter afle...

Watch this Scientific Journal Video about Murine Short Axis Ventricular Heart Slices for Electrophysiological Studies at JoVE.com

Murine Short Axis Ventricular Heart Slices for Electrophysiological Studies

Her beskriver vi forberedelsen af ​​levedygtige ventrikulære skiver fra voksne mus og deres anvendelse til skarpe elektrode-actionpotentialer. Disse multicellulære præparater tilvejebringer en bevaret in vivo- lignende vævsstruktur, hvilket gør dem til en værdifuld model for elektrofysiologiske og farmakologiske undersøgelser in vitro .

Murine Short Axis Ventricular Heart Skiver til Electrophysiological Studies

Murine cardiomyocytes have been extensively used for in vitro studies of cardiac physiology and new therapeutic strategies. However, multicellular ...

Research

JoVE Journal

Medicine

Reduction in Left Ventricular Wall Stress and Improvement in Function in Failing Hearts using Algisyl-LVR

Injection of  Algisyl-LVR, a treatment under clinical development, is intended to treat  patients with dilated cardiomyopathy. This treatment was recently ...

Murine Model of Wound Healing

Musemodel af Sårheling

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

Muse Livmoderhalskræft Heart transplantation model ved hjælp af en modificeret Cuff Teknik

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are ...

Intramyocardial Cell Delivery: Observations in Murine Hearts

Intramyocardial Cell Levering: Observationer i murine Hjerter

Previous studies showed that cell delivery promotes cardiac function amelioration by release of cytokines and factors that increase cardiac tissue ...

Murine Heterotopic Heart Transplant Technique

Murine Heterotop Heart Transplant Teknik

It is now over forty years since this technique was first reported by Corry, Wynn and Russell. Although it took some years for other labs to become ...

Utilizing Murine Inducible Telomerase Alleles in the Studies of Tissue Degeneration/Regeneration and Cancer

Ved hjælp Murin Inducerbar Telomerase Alleler i studier af væv Degeneration / Regeneration og Kræft

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that ...

Use of Two Intracorporeal Ventricular Assist Devices As a Total Artificial Heart

Brug af to Intracorporeal ventrikulær Assist enheder som en Total kunstige hjerteklapper

Mechanical circulatory support (MCS) has been introduced as a viable alternative to heart transplantation primarily through the use of intracorporeal ...

Electrophysiological Assessment of Murine Atria with High-Resolution Optical Mapping

Elektrofysiologiske vurdering af Murine forkamre med højopløselig optisk kortlægning

Recent genome-wide association studies targeting atrial fibrillation (AF) have indicated a strong association between the genotype and ...

Optocardiography and Electrophysiology Studies of Ex Vivo Langendorff-perfused Hearts

Optokardiografi og Elektrofysiologi studier af ex vivo Langendorff-perfbrugte hjerter

Small animal models are most commonly used in cardiovascular research due to the availability of genetically modified species and lower cost compared to ...

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

Murine Precision-Cut Lever skiver som en Ex Vivo Model af leverbiologi

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic ...