Name: ultrasound imaging-guided intracardiac injection to develop a mouse model of breast cancer brain metastases followed by longitudinal mri
Uploaded: 2014-03-06T13:45:00
Description: Watch this Scientific Journal Video about Ultrasound Imaging-guided Intracardiac Injection to Develop a Mouse Model of Breast Cancer Brain Metastases Followed by Longitudinal MRI at JoVE.com

Vi er taknemmelige for Drs. Diane Palmieri og Patricia Steeg af NCI for at give os MDA-MB231Br celler. Vi takker Dr. Ralph Mason, Mr. Jason Reneau og Ms Ramona Lopes til teknisk og kollegial støtte. Dette arbejde blev støttet delvist af DOD IDEA Awards W81XWH-08-1 til 0583 og W81XWH-12-1 til 0317. MRI eksperimenter blev udført i Advanced Imaging Research Center, en NIH BTRP # P41-RR02584 rum og ultralyd-vejledt intrakardiel injektion blev udført med VisualSonics Vevo 770 under 1S10RR02564801.

Alle animalske procedurer blev godkendt af Institutional Animal Care og brug Udvalg University of Texas Southwestern Medical Center. 1.. Forberedelse af MDA-MB231/Br-GFP Cells <ol><li> Hente og dyrke MDA-MB231/Br-GFP celler (venligst leveret af Drs. Palmieri og Steeg, NCI) i DMEM medium indeholdende 10% FBS, 1% glutamin og 1% penicillin / streptomycin. </li><li> Overhold tilstanden af ​​celler og farve af medium og erstatte medium hver 2-3 dage. </li><li> Trypsinisér og indsamle celle...

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I nærværende undersøgelse har vi vist, at ultralydsscanning-vejledt venstre ventrikel injektion sikrer nøjagtighed, således at alle dyr i denne undersøgelse udviklet hjernemetastaser og ingen dyr død blev observeret. Skønheden i image-vejledt injektion er, kan overvåges og justeres under billedet, som er forskellig fra den manuelle procedure, der kræver strenge anatomiske landmærker at følge stien for nål penetration af huden og til sidst ind i den venstre ventrikel. Aktuelle fo...

Hjernemetastaser er den mest almindelige intrakraniel malignitet hos voksne. Prognosen er yderst ringe, med en median overlevelse på 4-6 måneder, selv med aggressiv behandling. Brystkræft er en af de tre store primære kræftformer med en høj sygelighed af hjernemetastaser 1-3. Adskillige brystkræft hjerne-tropisk linjer er i stand til at udvikle hjernemetastaser efter intrakardial eller intracarotid injektion 4. Direkte injektion af tumorceller i den venstre ventrikel kan omgå lunge kapillarleje og dermed øge forekomsten af ​​dannelse af hjernemetastaser samtidig minimere viscerale metastaser. MDA-MB231Br linje er et af de mest anvendte humane brystkræft linjer til at udvikle hjernemetastaser i gnavermodeller 5,6. Ligesom mange andre undersøgelser 4,7, har vi foretaget en manuel procedure intrakardial indsprøjtning i vores tidligere undersøgelser. Imidlertid blev kun 50% succes, der opnås med den manuelleindsprøjtning og en brøkdel af mus døde af de gentagne invasive procedurer, hvis tidligere forsøg mislykkedes. Her indfører vi anvendelse af en imaging-styret procedure for at sikre injektion af hjerne-søger brystcancerceller i den venstre ventrikel af athymiske mus. Longitudinal høj opløsning MRI anvendes til at følge intrakraniel udvikling af hjernemetastaser.

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			<td height="21" style="height:21px;width:413px;">DMEM</td>
			<td style="width:136px;">HyClone</td>
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			<td height="21" style="height:21px;width:413px;">VisualSonics Vevo 770 High-Resolution Imaging System </td>
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			<td height="21" style="height:21px;">9.4T horizontal bore magnet with a Varian INOVA Unity system </td>
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			<td>Sakura Finetek USA</td>
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ultrasound imaging-guided intracardiac injection to develop a mouse model of breast cancer brain metastases followed by longitudinal mri

Brystkræft hjernemetastaser, der forekommer hos 30% af brystkræftpatienter på trin IV, er forbundet med høj dødelighed. Den mediane overlevelse er kun 6 måneder. Det er vigtigt at have egnede dyremodeller til at efterligne hæmodynamisk spredning af metastatiske celler i den kliniske scenario. Her bliver vi indføre brugen af ​​små dyr ultralydsscanning til at guide en nøjagtig indsprøjtning af hjerne tropisk brystkræftceller ind i venstre hjertekammer af athymiske nøgne mus. Longitudinal MRI bruges til at vurdere intrakraniel initiering og vækst af metastaser i hjernen. Ultralyd-vejledt intrakardial injektion sikrer ikke kun en nøjagtig injektion og herved en højere vellykket sats, men også signifikant reduceret dødelighed sammenlignet med vores tidligere manuel procedure. In vivo høj opløsning MRI tillader visualisering af hyperintense multifokale læsioner, så små som 310 um i diameter på T2-vægtede billeder på 3 uger efter injektion. Follow-up MRI afslører intrakraniel tumor vækst og øget antal af metastaser, der distribuerer hele hjernen.

Med den høje rumlige opløsning MRI (78 um i plan opløsning), kan hyperintense læsioner identificeres så lille som 310 um i diameter (figur 2). Da metastaser i denne undersøgelse er meget lille og udviklingen af nekrose og ødem er minimal, det hyperintens læsion på T 2-vægtede billeder virkelig repræsenteret tumormassen. Langsgående MRI undersøgelser tillader in vivo noninvasive evaluering af tumorvækst. Som vist i figur 3, h...

Watch this Scientific Journal Video about Ultrasound Imaging-guided Intracardiac Injection to Develop a Mouse Model of Breast Cancer Brain Metastases Followed by Longitudinal MRI at JoVE.com

Ultrasound Imaging-guided Intracardiac Injection to Develop a Mouse Model of Breast Cancer Brain Metastases Followed by Longitudinal MRI

En brystkræft hjernemetastaser musemodel etableres med ultralydsscanning-vejledt intrakardiel injektion af MDA-MB231/Br-GFP celler. Udvikling af multifokale intrakranielle metastaser er blevet overvåget i længderetningen ved hjælp af høj opløsning 9,4 T MR-scanning.

Ultrasound Imaging-vejledt intrakardiel Injection at udvikle en musemodel af Breast Cancer hjernemetastaser Efterfulgt af Longitudinal MRI

Breast cancer brain metastasis, occurring in 30% of breast cancer patients at stage IV, is associated with high mortality. The median survival is only 6 ...

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