Name: bioluminescent orthotopic model of pancreatic cancer progression
Uploaded: 2013-06-28T13:00:00
Description: Watch this Scientific Journal Video about Bioluminescent Orthotopic Model of Pancreatic Cancer Progression at JoVE.com

Dette arbejde blev støttet af National Sundhed og Medical Research Council, Australien (1008865), den australske forskningsråd (LE110100125), National Cancer Institute (CA138687-01), Erica Sloan er understøttet af en tidlige karriere Fellowship fra National Breast Cancer Foundation, Australien. Corina Kim-Fuchs er understøttet af et stipendium fra den schweiziske Cancer League og et HDR stipendium fra Monash Institute of Pharmaceutical Sciences. Eliane Angst er støttet af en bevilling fra Bern Cancer League.

Protokollen bliver demonstreret udføres under vejledning og godkendelse af forfatterens institutionens Dyrepleje og anvendelse udvalg. Alle forsøg udføres i overensstemmelse med alle relevante retningslinjer, regler og reguleringsorganer. 1.. Transduktion bugspytkirtelkræft Cell Lines <ol><li> Transducere bugspytkirtelkræftceller at udtrykke luciferase som tidligere beskrevet. 12,13 Panc-1 og Capan-1 bugspytkirtelkræft cellelinjer transduceret med ildflueluciferase anvende...

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Her beskriver vi en ortotopisk model til langsgående vurdering af pancreas tumor udvikling og progression. Primær tumor vækstkinetik er reproducerbare (figur 3) og kan være non-invasivt overvåges ved hjælp af bioluminescens billeddannelse af luciferase-mærkede celler, fx til analyser af tumor respons på nye anti-bugspytkirtlen kræftmedicin. I overensstemmelse med den humane sygdom, viser modellen lokale pancreas invasion (figur 4A), som muliggør undersøgelse af tumor...

Kræft i bugspytkirtlen er den fjerde hyppigste årsag til kræft-relaterede dødsfald, med en 5-års overlevelse på 4-6%. 1,2 Kun 15% af patienterne er diagnosticeret tidligt nok i sygdommen for at være berettiget til operation og tumorer gentages i&gt; 80% af de patienter. 3,4 Gemcitabin anvendes til behandling af pancreas-adenocarcinomer, dog kemoterapiresistens er almindelig og ofte lægemidlet har ringe indvirkning på den samlede overlevelse. 5 nye farmakologiske strategier til behandling bugspytkirtelkræft kritisk behov. Deres udvikling afhænger væsentligt forbedret forståelse af de vigtigste trin i sygdomsprogression, der kan være følsomme over for terapeutisk intervention. Ortotopisk modeller af kræft i bugspytkirtlen efterligne centrale aspekter af den menneskelige sygdomme, hvilket gør dem ideelle værktøjer til at studere biologi af kræft i bugspytkirtlen. 6-9 I modsætning til in vitro cellebaserede analyser af kræft i bugspytkirtlen celle adfærd end subkutan in vivo modeller for kræft i bugspytkirtlen, ortotopisk modeller tillader efterforskning af tumor celle interaktioner med bugspytkirtlen mikromiljø. Kinetik sygdomsprogression er meget reproducerbar i ortotopisk modeller, og sker over en kort tidsramme (uger), hvilket gør dem velegnede til at pre-klinisk afprøvning af nye lægemidler. Dette er i modsætning til transgene modeller, hvor sygdomsstart sker over en længere og mere variabel tidsramme (måneder til 1 år). 10. Når det bruges med mere aggressive cellelinier, ortotopisk modeller af bugspytkirtelkræft har mønstre af spontan metastase svarende til dem set i patienter. 8. Angivelse af bioluminiscerende reportergener såsom ildflueluciferase letter langsgående overvågning af tumorvækst, metastatisk spredning, tilbagefald og respons på lægemidler. 6,11 Her beskriver vi en ortotopisk model af kræft i bugspytkirtlen, der udnytter MatrIgel for lokal celle levering og in vivo bioluminescens billeddannelse til ikke-invasiv overvågning af tumorprogression. Denne ortotopisk model af kræft i bugspytkirtlen giver ikke-invasive analyser af sygdommen og reaktion på terapeutiske interventioner i syngene eller xenograft modeller.

<table border="1">
 <tbody>
 <tr>
 <td>Equipment</td>
 <td>Company</td>
 <td>Catalog Number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Clean Bench coat</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
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 <td>Heating pad</td>
 <td></td>
 <td></td>
 <td>Set to 37 &deg;C </td>
 </tr>
 <tr>
 <td>Ivis Lumina ll Bioluminescent imager</td>
 <td>Caliper </td>
 <td></td>
 <td>Alternative bioluminescent imaging systems include In vivo F PRO (Carestream) and Photon Imager (Biospace Lab)</td>
 </tr>
 <tr>
 <td>Dissecting scissors</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Iris forceps (serrated)</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Needle holder</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>27 G 0.3 ml insulin syringe</td>
 <td>Terumo</td>
 <td>T35525M2913</td>
 <td></td>
 </tr>
 </tbody>
</table>

bioluminescent orthotopic model of pancreatic cancer progression

<p class="jove_content"&gt; Bugspytkirtelkræft har en meget dårlig fem-års overlevelse på 4-6%. Nye terapeutiske muligheder er kritisk behov og afhænger forbedret forståelse af kræft i bugspytkirtlen biologi. For bedre at forstå interaktionen af ​​kræftceller med pancreas mikromiljø viser vi en ortotopisk model af bugspytkirtelkræft, der tillader ikke-invasiv overvågning af udviklingen af ​​kræft. Luciferase-mærkede bugspytkirtelkræftceller resuspenderes i Matrigel og leveret i bugspytkirtlen halen under laparotomi. Matrigel størkner ved kropstemperatur for at forhindre lækage af cancerceller under indsprøjtning. Primær tumorvækst og metastase til fjerne organer overvåges efter injektion af luciferasesubstrat luciferin hjælp<em&gt; In vivo</em&gt; Billeddannelse af bioluminescens emission fra kræftcellerne.<em&gt; In vivo</em&gt; Billeddannelse kan også bruges til at spore primær tumor tilbagefald efter resektion. Denne ortotopisk model er velegnet til både syngene og xenograft modeller, og kan anvendes i prækliniske forsøg for at undersøge virkningen af ​​nye kræftlægemidler på væksten i den primære pancreas tumor og metastaser.</p&gt;

Denne metode beskriver en ortotopisk model af bugspytkirtelkræft hjælp kirurgiske procedurer, herunder induktion af anæstesi, laparotomi injektion af cancerceller i Matrigel og abdominal lukning (fig. 1A). De injicerede celler danner en boble i overfladen af bugspytkirtlen (figur 1B). Bugspytkirtelkræft progression kan være ikke-invasivt overvåges ved hjælp af in vivo bioluminescens imaging at spore cancercelleproliferation og formidling (figur 2). Lever...

Watch this Scientific Journal Video about Bioluminescent Orthotopic Model of Pancreatic Cancer Progression at JoVE.com

Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

Forbedret forståelse af kræft i bugspytkirtlen biologi kritisk nødvendige for, at udviklingen af ​​bedre terapeutiske muligheder for at behandle kræft i bugspytkirtlen. At imødekomme dette behov, viser vi en ortotopisk model af bugspytkirtelkræft, der tillader ikke-invasiv overvågning af udviklingen af ​​kræft ved hjælp af<em&gt; In vivo</em&gt; Bioluminescens billeddannelse.

Bioluminescent ortotopisk Model af kræft i bugspytkirtlen Progression

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%.  New therapeutic options are critically needed and depend on improved ...

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