Name: imaging glioma initiation in vivo through a polished and reinforced thin-skull cranial window
Uploaded: 2012-11-20T12:00:00
Description: Watch this Scientific Journal Video about Imaging Glioma Initiation In Vivo Through a Polished and Reinforced Thin-skull Cranial Window at JoVE.com

Dette arbeidet er støttet av Jackson Laboratory Cancer Center Pilot Grant og The Maine Cancer Foundation.

1. Protokoll <ol><li> Anesthetize musen med ketamin og xylazin i en dose på 0,1 mg ketamin og 0,01 mg av xylazin per 1 g kroppsvekt. Karprofen (0.005 mg per 1 g kroppsvekt) benyttes til analgesi og administreres preoperativt. </li><li> Alle kirurgiske verktøy, inkludert dental borkronen, er dampsteriliseres i en autoklav. Hvis batch operasjoner skal utføres, må spissene på kirurgiske instrumenter steriliseres med et glass bead sterilizer (Fine Science Verktøy FST 250) før hver påfølgende operasjon. </li><li> ...

<ol>
	<li>Paulino, A. C., Teh, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+brain+tumors.">Treatment of brain tumors.</a> N. Engl. J. Med. 352, 2350-2353 (2005).</li><li>Stupp, R., 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=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>Singh, S. 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=Identification+of+human+brain+tumour+initiating+cells.">Identification of human brain tumour initiating cells.</a> Nature. 432, 396-401 (2004).</li><li>Hanahan, D., Weinberg, R. 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Nøkkelen til en vellykket porter cranial vindu er tynning og polering. Mens den første tynning kan utføres raskt, bør man sørge for å sikre homogen tynning av skallen over et stort område. Vi vanligvis bruke et tynt lag av saltvann til skallen, og deretter tynn skallen ett pass gangen, slik at saltoppløsningen fordamper kort etter microdrill har passert over skallen gang. Dette gir oss muligheten til å sakte men homogent ytterligere tynn skallen. En stødig hånd under mikroskopet er også viktig. Når skull-ty...

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imaging glioma initiation in vivo through a polished and reinforced thin-skull cranial window

Glioma er en av de mest dødelige former for menneskelig kreft. Den mest effektive glioma terapi til dato-kirurgi etterfulgt av strålebehandling-gir pasientene beskjedne fordeler, som de fleste pasienter ikke overleve mer enn fem år etter diagnose på grunn av glioma tilbakefall 1,2. Oppdagelsen av kreft stamceller i menneskelige hjernesvulster holder løftet for å ha en enorm innvirkning på utviklingen av nye terapeutiske strategier for glioma 3. Kreftstamceller defineres ved sin evne både til selv fornye og å skille, og er tenkt å være de eneste celler i en svulst som har kapasitet til å initiere nye svulster 4. Glioma tilbakefall etter strålebehandling antas å oppstå fra motstand av glioma stamceller (GSCS) til terapi 5-10. In vivo, er GSCS vist seg å ligge i en perivaskulær nisje som er viktig for å opprettholde sine stamcelle-lignende egenskaper 11-14 . Sentralt i den organisasjonensjon av GSC nisje er vaskulære endotelceller 12. Eksisterende bevis tyder på at GSCS og deres samspill med vaskulære endotelceller er viktig for tumor utvikling, og identifisere GSCS og deres samspill med endotelceller som viktige terapeutiske mål for glioma. Tilstedeværelsen av GSCS bestemmes eksperimentelt ved deres evne til å initiere nye svulster ved orthotopic transplantasjon 15. Dette er vanligvis oppnås ved å injisere et bestemt antall GBM celler isolert fra humane svulster i hjernen på alvorlig immun-mangelfull mus, eller av mus GBM celler i hjernen på congenic vert mus. Analyser for tumorvekst blir deretter utføres etter tilstrekkelig tid til å tillate GSCS blant injiserte GBM cellene til å gi opphav til nye svulster-typisk flere uker eller måneder. Derfor trenger eksisterende analysene ikke tillate undersøkelse av viktige patologisk prosess startfasen fra enkle GSCS in vivo. Derfor viktig insights i de spesifikke roller GSCS og deres samspill med vaskulære endotelceller i de tidlige stadier av startfasen mangler. Slike innsikter er avgjørende for å utvikle nye terapeutiske strategier for gliom, og vil ha store konsekvenser for å hindre glioma tilbakefall hos pasienter. Her vi har tilpasset Portene cranial vindu prosedyre 16 og in vivo to-foton mikroskopi å tillate visualisering av startfasen fra injisert GBM celler i hjernen av en levende mus. Vår teknikk vil bane vei for fremtidige arbeidet med å belyse de viktigste signaliserer mekanismer mellom GSCS og vaskulære endotelceller under glioma innvielse.

Watch this Scientific Journal Video about Imaging Glioma Initiation In Vivo Through a Polished and Reinforced Thin-skull Cranial Window at JoVE.com

Imaging Glioma Initiation In Vivo Through a Polished and Reinforced Thin-skull Cranial Window

Ved å kombinere en polert og forsterket tynn-skull (porter) cranial vinduet og glioblastoma (GBM) celle injeksjon, kan vi observere glioma initiering og vekst fra injiserte GBM celler i hjernen til en levende mus i lengderetningen.

Imaging Glioma Initiation<em&gt; In Vivo</em&gt; Gjennom en polert og Forsterket Thin-skull Cranial Window

Glioma is the one of the most lethal forms of human cancer. The most effective glioma therapy to date-surgery followed by radiation treatment-offers ...

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Imaging Glioma Initiation In Vivo Through a Polished and Reinforced Thin-skull Cranial Window

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