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 taknemmelige for Dr. Andrew Hollander, Sara Davis, Lee Shuman, Tim Jenkins, og Dr. Xiangsheng Xu for deres eksperthjælp. Vi anerkender støtten fra Dr. Ann Kennedy. BCB blev støttet på Radiation Biology Training Grant C5T32CA009677. JFD blev støttet på Burroughs Wellcome Career Award for medicinske forskere (1.006.792). JLB blev støttet på supers tilskud (5 R25 CA140116-03). Vi vil gerne anerkende Dr. Steve Hahn hvis opmuntring og støtte har bidraget til at gøre vores forskning mulig. Vi vil også gerne takke University of Pennsylvania Nano-Bio-grænseflade Center (NBIC) og Dr. Dennis Discher for opmuntring og nyttige kommentarer. Vi anerkender Small Animal Imaging Facility (SAIF) ved University of Pennsylvania for brug af deres MRI og optisk / Bioluminescence Core Facilities. Disse teknikker blev udviklet som en del af projekter, der blev støttet af National Institutes of Health (RC1 CA145075 og K08 NS076548-01).

A. Pre-Operative tumorcelle Preparation <ol><li> Transducere U251 glioblastoma multiforme-celler med en lentiviral ekspressionsvektor (pGreenFire, System Biosciences) stabilt at udtrykke ildflue-luciferase-genet. </li><li> Disse celler blev dyrket i 10 ml komplet Dulbeccos modifikation af Eagles medium (DMEM), der består af DMEM suppleret med 10% føtalt kalveserum, 1% penicillin-streptomycin og 1% ikke-essentielle aminosyrer i en T75 vævsdyrkningskolbe inkubering ved 5% C0 2 og 37 ° C. </li><li> Udføre...

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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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Fremgangsmåden til stereotaktisk implantation af cancerceller i mus er beskrevet i dette dokument reproducerbart frembringer tumorer, som med rimelighed rekapitulere den infiltrativ og hurtig vækst mønster af klinisk glioblastoma multiforme 2, 6-8. Denne teknik er specielt velegnet til eksperimenter stratificere mus jævnt til forskellige behandlingsgrupper, hvor reproducerbare tumorer af sammenlignelig størrelse og biologiske egenskaber og i specifikke anatomiske steder er ønskelige. Stereotaktisk impla...

<table border="1"><tbody><tr><td> Beskrivelse </td><td> Leverandør </td><td> Katalognummer </td><td> Kommentarer </td></tr><tr><td> Digital Bare for Mouse stereotaktisk Instrument </td><td> Stoelting </td><td> 51730D </td><td> Stereotaktisk platform for mus implantation </td></tr><tr><td> Ketamin / xylazin </td><td></td><td></td><td> Injicerbar anæstesi </td></tr><tr><td> Puralube Vet Salve (oftalmiske) </td><td> Amazon.com </td><td></td><td> For at forhindre udtørring af musens øjne </td></tr><tr><td> bore holder til stereotaktisk platform </td><td> Stoelting </td><td> 51.681 </td><td></td></tr><tr><td> Mikromotoren Electric Drill </td><td> Stoelting </td><td> 51.449 </td><td> Til boring gennem kraniet </td></tr><tr><td> 0,45 mm carbide borekronen </td><td> Stoelting </td><td> 514.551 </td><td></td></tr><tr><td> Sterile vatsvabere </td><td> Fisher Scientific </td><td> 23-400-100 </td><td></td></tr><tr><td> Glas perle tør sterilisator (Germinator 500) </td><td> Braintree Scientific </td><td> GER-5287 </td><td> At sterilisere metal kirurgiske instrumenter </td></tr><tr><td> Mus rektal sonde </td><td> Braintree Scientific </td><td> RET-3-ISO </td><td> Kompatibel med temperatur regulatoren </td></tr><tr><td> Temperatur Controller (TCAT-2DF) </td><td> Harvard Apparatus </td><td> 727.561 </td><td> Temperaturregulator at opretholde dyrets temperatur under kirurgi </td></tr><tr><td> Lille varmeplade </td><td> Harvard Apparatus </td><td> 727.617 </td><td> Til brug med temperaturregulator at varme mus under kirurgi. Varmepladen passer under musen på den stereotaxiske platform. </td></tr><tr><td> Engangs Skalpeller </td><td> BD Bard-Parker </td><td> 2015-11 </td><td> # 10 skalpel </td></tr><tr><td> 10 mikroliter sprøjte </td><td> Hamilton </td><td> 7635-01 </td><td> Til injektion af tumorceller </td></tr><tr><td> 30 gauge nåle, 1 &quot;lange, med flad punkt </td><td> Hamilton </td><td> Various </td><td> Skal være kompatibel med 10 pi sprøjte </td></tr><tr><td> Nanomite Programmerbar Sprøjtepumpe </td><td> Harvard Apparatus </td><td> 704.507 </td><td> Digital motoriseret sprøjte injektor til stereotaktisk anordning </td></tr><tr><td> Cellulose sterile kirurgiske spyd svampe </td><td> UltraCell </td><td> 40.410 </td><td> At tørre den kirurgiske område </td></tr><tr><td> Knoglevoks </td><td> Ethicon </td><td> W31 </td><td> At tætne borehullet </td></tr><tr><td> Tissumend II syntetisk absorberbar vævsklæbemiddel </td><td> Veterinær Products Laboratories </td><td> 3002931 </td><td> At forsegle incisionen </td></tr><tr><td> Varmt vand pumpe med opvarmning pad </td><td> Gaymar </td><td> TP-650 </td><td> Varmer mus i postoperative periode </td></tr><tr><td> IVIS Lumina II </td><td> Caliper Life Science </td><td></td><td> Bioluminescent imager </td></tr><tr><td> D-luciferin kaliumsalt </td><td> Guld Bioteknologi </td><td> LUCK-1 </td><td> Luciferin for bioluminescerende billeddannelse </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 high-grade primær kræft i hjernen med en median overlevelse på kun 14,6 måneder hos mennesker trods standard tri-modalitet behandling bestående af kirurgisk resektion, postoperativ strålebehandling og temozolomid kemoterapi 1. Nye terapeutiske tilgange er klart behov for at forbedre patienternes overlevelse og livskvalitet. Udviklingen af ​​mere effektive behandlingsstrategier vil blive hjulpet af dyremodeller for GBM der rekapitulere sygdom hos mennesker endnu tillade seriel billeddannelse til at overvåge tumor vækst og behandlingsrespons. I denne artikel beskriver vi vor teknik til præcis stereotaktisk implantation af bio-billedområdet GBM kræftceller i hjernen på nøgne mus resulterer i tumorxenotransplantater at rekapitulere vigtige kliniske træk ved GBM 2. Denne metode giver tumorer, som er reproducerbare og er placeret i præcise anatomiske steder, samtidig med at in vivo bioluminescerende billeddannelse til serielt overvåge intrakraniel xenotransplantat vækst og respons på behandling 3-5. Denne metode er også godt tolereret af dyrene med lav perioperative sygelighed og dødelighed.

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 integreret fremgangsmåde til præcis stereotaktisk implantation af humane glioblastoma multiforme-celler ind i hjernen på nøgne mus og efterfølgende serielle<em&gt; In vivo</em&gt; Billedbehandling til at overvåge vækst og reaktion på behandlingen af ​​de resulterende xenotransplantater.

Stereotaktisk Intrakraniel Implantation og<em&gt; In vivo</em&gt; Bioluminescent Imaging af tumorvæv implanteret i en musemodel System af 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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