Name: stereotactic intracranial implantation and in vivo bioluminescent imaging of tumor xenografts in a mouse model system of glioblastoma multiforme
Uploaded: 2012-09-25T11:00:00
Description: Watch this Scientific Journal Video about Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme at JoVE.com

Vi er takknemlige for Dr. Andrew Hollander, Sara Davis, Lee Shuman, Tim Jenkins, og Dr. Xiangsheng Xu for deres eksperthjelp. Vi erkjenner støtte fra Dr. Ann Kennedy. BCB ble støttet på Radiation Biology Training Grant C5T32CA009677. JFD ble støttet på Burroughs Wellcome Career Award for medisinske forskere (1.006.792). JLB ble støttet på Supers stipend (5 R25 CA140116-03). Vi ønsker å erkjenne Dr. Steve Hahn som oppmuntring og støtte har bidratt til å gjøre vår forskning mulig. Vi vil også gjerne takke University of Pennsylvania Nano-Bio Interface Center (NBIC) og Dr. Dennis Discher for oppmuntring og nyttige kommentarer. Vi erkjenner Small Animal Imaging Facility (Saif) ved University of Pennsylvania for bruk av deres MR og optisk / Bioluminescens kjernefasiliteter. Disse teknikkene ble utviklet som en del av prosjektene som ble støttet av National Institutes of Health (RC1 CA145075 og K08 NS076548-01).

A. preoperativ Tumor Cell Forberedelse <ol><li> Transduce U251 glioblastoma multiforme celler med en lentiviral uttrykk vektor (pGreenFire, System biovitenskap) til stabilt uttrykke firefly luciferase genet. </li><li> Disse cellene ble dyrket i 10 ml komplett Dulbeccos Modifikasjon av Eagles medium (DMEM), som består av DMEM supplert med 10% føtalt kalveserum, 1% penicillin-streptomycin, og 1% ikke-essensielle aminosyrer i en T75 vevskultur kolbe inkubering ved 5% C0 2 og 37 ° C. </li><li> Utfør standar...

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	<li>Stupp, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiotherapy+plus+concomitant+and+adjuvant+temozolomide+for+glioblastoma.">Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma.</a> N. Engl. J. Med. 352, 987-996 (2005).</li><li>Jacobs, V. L., Valdes, P. A., Hickey, W. F., De Leo, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+review+of+in+vivo+GBM+rodent+models:+emphasis+on+the+CNS-1+tumour+model.">Current review of in vivo GBM rodent models: emphasis on the CNS-1 tumour model.</a> ASN Neuro. 3, e00063 (2011).</li><li>Shelton, L. 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Metoden for stereotactic implantasjon av kreftceller hos mus som er beskrevet i denne artikkelen genererer reproduserbart svulster som rimelig rekapitulere infiltrerende og rask vekst mønster av kliniske glioblastoma 2 multiforme, 6-8. Denne teknikken er spesielt godt egnet til eksperimenter stratifying mus jevnt til forskjellige behandlingsgrupper der reproduserbare svulster av sammenlignbar størrelse og biologiske egenskaper og i bestemte anatomiske steder er ønskelig. Stereotactic implantasjon av kreftc...

<table border="1"><tbody><tr><td> Beskrivelse </td><td> Leverandør </td><td> Katalognummer </td><td> Kommentarer </td></tr><tr><td> Digital Just for Mouse stereotaksiske Instrument </td><td> Stoelting </td><td> 51730D </td><td> Stereotactic plattform for mus implantasjon </td></tr><tr><td> Ketamin / xylazin </td><td></td><td></td><td> Injiserbart anestesi </td></tr><tr><td> Puralube Vet salve (ophthalmic) </td><td> Amazon.com </td><td></td><td> Å forhindre uttørking av musens øyne </td></tr><tr><td> bore holder for stereotactic plattformen </td><td> Stoelting </td><td> 51681 </td><td></td></tr><tr><td> Micromotor elektrisk drill </td><td> Stoelting </td><td> 51449 </td><td> For boring gjennom skallen </td></tr><tr><td> 0,45 mm karbid drill bit </td><td> Stoelting </td><td> 514551 </td><td></td></tr><tr><td> Sterile bomullspinner </td><td> Fisher Scientific </td><td> 23-400-100 </td><td></td></tr><tr><td> Glass perle tørr sterilisator (Germinator 500) </td><td> Braintree Scientific </td><td> GER-5287 </td><td> Å sterilisere metall kirurgiske instrumenter </td></tr><tr><td> Mus rektal probe </td><td> Braintree Scientific </td><td> RET-3-ISO </td><td> Kompatibel med temperaturregulatoren </td></tr><tr><td> Temperatur Controller (TCAT-2DF) </td><td> Harvard Apparatus </td><td> 727561 </td><td> Temperaturkontroller for å opprettholde dyrets temperatur under operasjonen </td></tr><tr><td> Liten oppvarming plate </td><td> Harvard Apparatus </td><td> 727617 </td><td> For bruk med temperaturregulator å varme mus under operasjonen. Varmeplaten passer under musen på stereotaksiske plattformen. </td></tr><tr><td> Disponibel Skalpeller </td><td> BD Bard-Parker </td><td> 2015-11 </td><td> # 10 skalpell </td></tr><tr><td> 10 mikroliter sprøyte </td><td> Hamilton </td><td> 7635-01 </td><td> For injeksjon av kreftceller </td></tr><tr><td> 30 gauge nåler, en &quot;lang, med flat punkt </td><td> Hamilton </td><td> Various </td><td> Må være kompatibelt med den 10 pl sprøyten </td></tr><tr><td> Nanomite Programmerbar Sprøytepumpe </td><td> Harvard Apparatus </td><td> 704507 </td><td> Digital motorisert sprøyte injektor for stereotaksiske enhet </td></tr><tr><td> Cellulose sterile kirurgiske spyd svamper </td><td> Ultracell </td><td> 40410 </td><td> Å tørke den kirurgiske feltet </td></tr><tr><td> Bone voks </td><td> Ethicon </td><td> W31 </td><td> Å forsegle graden hull </td></tr><tr><td> Tissumend II syntetisk absorberbare vev lim </td><td> Veterinærprodukter Laboratories </td><td> 3002931 </td><td> Å forsegle snittet </td></tr><tr><td> Varmt vann pumpe med oppvarming pad </td><td> Gaymar </td><td> TP-650 </td><td> Varmer mus i postoperative perioden </td></tr><tr><td> IVIS Lumina II </td><td> Målepunktet Life Science </td><td></td><td> Bioluminescent imager </td></tr><tr><td> D-Luciferin kaliumsalt </td><td> Gull Bioteknologi </td><td> LYKKE-1 </td><td> Luciferin for bioluminescent bildebehandling </td></tr></tbody></table>

stereotactic intracranial implantation and in vivo bioluminescent imaging of tumor xenografts in a mouse model system of glioblastoma multiforme

Glioblastoma multiforme (GBM) er en høyverdig primære kreft i hjernen med en median overlevelse på bare 14,6 måneder hos mennesker tross standard tri-modalitet behandling bestående av kirurgisk reseksjon, postoperativ strålebehandling og Temozolomide kjemoterapi en. Nye terapeutiske tilnærminger er klart behov for å forbedre pasientens overlevelse og livskvalitet. Utviklingen av mer effektive behandlingsstrategier ville bli hjulpet av dyremodeller av GBM som rekapitulere sykdom hos mennesker, men la seriell imaging å overvåke svulsten vekst og behandlingsrespons. I denne artikkelen beskriver vi vår teknikk for nøyaktig stereotactic implantasjon av bio-bildeområdet GBM kreftceller i hjernen på nakne mus som resulterer i tumorxenotransplantater som rekapitulere viktige kliniske funksjoner i GBM 2. Denne metoden gir svulster som er reproduserbare og er lokalisert i presise anatomiske steder samtidig som in vivo bioluminescent imaging til serielt overvåke intrakranial xenograft vekst og respons til behandlinger 3-5. Denne metoden er også godt tolerert av dyr med lav perioperativ morbiditet og mortalitet.

Watch this Scientific Journal Video about Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme at JoVE.com

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

Vi beskriver en integrert metode for den presise, stereotactic Implantering av humant glioblastoma multiforme celler i hjernen på nakne mus og etterfølgende serienummer<em&gt; In vivo</em&gt; Imaging å overvåke vekst og respons på behandling av de resulterende xenografter.

Stereotactic Intrakraniell Implantasjon og<em&gt; In vivo</em&gt; Bioluminescent Imaging av tumorxenotransplantater i en mus modell System of glioblastoma multiforme

Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality ...

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Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

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