Name: ultrasound-guided transthoracic intramyocardial injection in mice
Uploaded: 2014-08-05T15:00:00
Description: Watch this Scientific Journal Video about Ultrasound-guided Transthoracic Intramyocardial Injection in Mice at JoVE.com

TWP was funded by the Irish Cardiac Society Brian McGovern Travelling Fellowship. WTP was funded by R01 HL095712 and an AHA Established Investigator Award.

Alle beskrevne trin blev udført under protokoller, der er godkendt af Institutional Animal Care og brug Udvalg Boston Børnehospital. 1. Fremstilling <ol><li> Udfør baseline hjerteundersøgelser anatomisk og funktionel vurdering af ekkokardiografi før begynder injektionsprotokollen, som den optimale faste transducer position intramyocardial injektion ikke kan give de optimale standard udsigt til afgrænsning anatomi og vurdering af funktion. </li><li> Setup injektion er vist i f...

<ol>
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Biologics kan leveres til myokardiet ved direkte intramyocardial injektion, intrapericardial injektion eller indirekte indgivelse via blodbanen. Nylige cellebaserede terapi forsøg i myokardieinfarkt modeller har beskrevet en åben torakotomi tilgang til levering af injektat 12-14. En vigtig faktor for succesen af ​​en myocardial terapeutisk intervention afhænger valget af levering rute. Den højeste lokale dosis biologisk opnås ved intramyocardial levering 15,16. Intramyocardial injektion un...

Hjertesygdom er den hyppigste dødsårsag for både mænd og kvinder i USA, der tegner sig for 600.000 dødsfald årligt 1. Murine modeller af hjerte-kar-sygdom er kritisk vigtige for at undersøge patofysiologiske mekanismer og for at udforske mulige behandlinger. Myocardial levering af genterapivektorer stamceller, modificerede RNA og andre terapeutiske midler tillader undersøgelse af deres terapeutiske potentiale for hjertesygdomme 2-7. I øjeblikket er der begrænsede muligheder for myokardie levering af terapeutiske midler i musemodeller 6. Intramyocardial injektion under direkte visualisering er almindeligt anvendt, men kræver en sternotomi eller torakotomi og er begrænset til det belyste område af hjertet. Mens praktisk, når udført på tidspunktet for en anden eksperimentel manipulation såsom LAD-ligering, der er behov for en invasiv procedure til intramyocardial levering begrænser potentielle eksperimentelle design og indfører enDERLIGERE effekter fra proceduren (f.eks fibrose skyldes thoracotomi). Perkutan perikardial levering af virale vektorer er blevet rapporteret, men stedet og distribution af terapeutisk middel ikke er homogen og er vanskelig at kontrollere 8. Perkutan koronar injektion resulterer i mere homogen fordeling af injicerede materiale, men effektiv og reproducerbar koronar levering er udfordrende i murine modeller. Her beskriver vi en lukket kiste intramyocardial indsprøjtning teknik, der gør det muligt for minimalt invasiv, operatør kontrolleret målretning af terapeutiske midler ved hjælp af ultralyd. Teknikken er let at lære, overflødiggør behovet for torakotomi eller sternotomi og deres ledsagende eksperimentelle komplikationer, og giver større fleksibilitet med hensyn til tidspunktet og steder af intramyocardial injektion. Således ekkokardiografi-assisteret intramyocardial injektion udgør en teknisk enkel og meget effektiv metode til at manipuleremyokardiet i murine eksperimentelle modeller.

<table border="0" cellpadding="0" cellspacing="0" width="743">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Vevo 2100 ultrasound imaging system&#160;</td>
			<td style="width:144px;">Visualsonics</td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Vevo Integrated Rail System III</td>
			<td style="width:144px;">Visualsonics</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Microscan MS400 transducer</td>
			<td style="width:144px;">Visualsonics</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Microscan MS550D transducer</td>
			<td style="width:144px;">Visualsonics</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">PrecisionGlide needles</td>
			<td style="width:144px;">BD</td>
			<td style="width:119px;">305128</td>
			<td>30G x 1 inch</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">1cc slip tip syringe</td>
			<td style="width:144px;">Exel International&#160;</td>
			<td style="width:119px;">26048</td>
			<td>or equivalent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Gastight 50ul glass syringes</td>
			<td style="width:144px;">Hamilton&#160;</td>
			<td style="width:119px;">1705</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Trypan blue stain (0.4%)</td>
			<td style="width:144px;">Gibco&#160;</td>
			<td style="width:119px;">15250</td>
			<td>or equivalent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Isoflurane</td>
			<td style="width:144px;">Baxter</td>
			<td style="width:119px;">AHN3640</td>
			<td>or equivalent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Aquasonic 100</td>
			<td style="width:144px;">Parker Laboratories</td>
			<td style="width:119px;">(01-08)</td>
			<td>or equivalent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Polystyrene microspheres (red fluorescent)</td>
			<td style="width:144px;">Life Technologies</td>
			<td style="width:119px;">F-8842</td>
			<td>or equivalent</td>
		</tr>
	</tbody>
</table>

ultrasound-guided transthoracic intramyocardial injection in mice

Murine modeller af hjerte-kar-sygdom er vigtige for at undersøge patofysiologiske mekanismer og udforske potentielle regenererende behandlinger. Eksperimenter med myokardie injektion i øjeblikket udføres af direkte kirurgisk adgang gennem en torakotomi. Mens praktisk, når udført på tidspunktet for en anden eksperimentel manipulation såsom kranspulsåreombinding, behovet for en invasiv procedure til intramyocardial levering begrænser potentielle eksperimentelle design. Med stadigt forbedre ultralyd opløsning og avancerede invasive billeddiagnostiske metoder, er det nu muligt at rutinemæssigt udføre ultralyd-vejledt perkutan intramyocardial injektion. Denne modalitet effektivt og pålideligt leverer midler til et målrettet region myokardiet. Fordele ved denne teknik omfatter undgåelse af kirurgisk morbiditet, mulighed for at målrette områder af myocardium selektivt ved hjælp af ultralyd, og muligheden for at levere injektionsmidlet til myocardium ved multiple, forudbestemte tidsintervaller. Med praktiseres teknik, komplikationer fra intramyocardial injektion er sjældne, og mus hurtigt vende tilbage til normal aktivitet på nyttiggørelse fra bedøvelsesmiddel. Efter trinene i denne protokol, kan operatøren med grundlæggende ekkokardiografi erfaring hurtigt blive kompetent i denne alsidige, minimalt invasiv teknik.

Murine intramyocardial Injektion med blåt farvestof eller fluorescerende mikrosfærer Injektion af Evans blåt farvestof er nyttig med henblik på uddannelse. Snart efter injektion, aflive mus og fjerne hjertet til at visualisere placeringen af den injicerede blåt farvestof. Figur 5 viser et eksempel på en succesfuld injektion med blåt farvestof infiltrerer myokardiet mid-papillære muskel niveau (figur 5A, område indesluttet den stiplede linje). Evans-bl?...

Watch this Scientific Journal Video about Ultrasound-guided Transthoracic Intramyocardial Injection in Mice at JoVE.com

Ultrasound-guided Transthoracic Intramyocardial Injection in Mice

Ekkokardiografi-guidede perkutan intramyocardial injektion er en effektiv, pålidelig og målrettes modalitet til levering af genoverførsel agenter eller celler i murine hjerte. Efter trinene i denne protokol, kan operatøren hurtigt blive kompetent i denne alsidige, minimalt invasiv teknik.

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