Name: an orthotopic mouse model of spontaneous breast cancer metastasis
Uploaded: 2016-08-14T13:00:00
Description: Watch this Scientific Journal Video about An Orthotopic Mouse Model of Spontaneous Breast Cancer Metastasis at JoVE.com

Supported by grants from the National Institute of Health grants CA133085, CA182518 and CA185909 (to KL) and VA Merit Review Award BX001962 (to KL).

Alle procedurer følger retningslinjer og godkendte protokoller af Georgia Regents University Animal brug og Omsorgsudvalget. 1. Etablering af ortotopisk Breast Cancer Tumor <ol><li> En dag før forsøget, kultur ca. 2 x 10 6 4T1-Luc tumorceller i 10 cm dyrkningsskål i 10 ml RPMI-medium indeholdende 10% FBS. Inkubér dyrkningsskålen i en CO2-inkubator ved 37 ° C og 5% CO2. </li><li> På dagen for tumorcelleinjektion, fjerne dyrkningsmediet fra dyrkningsskålen med vakuum, ti...

<ol>
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Der er blevet udviklet mange typer af transgene musemodeller for brystkræft metastase. 1 Disse transgene mus har høj tumorincidensen i området fra 60 til 100%. Imidlertid metastase incidens af disse transgene mus er meget lavere end den tumorincidensen (14 - 100%). Tumorceller metastaserer til LN og lunger i de fleste af disse transgene musemodeller for brystkræft metastase. Metastaser latenstid varierer fra model til model og varierer fra 2 til 8 måneder. 6,11-16 Disse transgene musemodeller ...

Ifølge American Cancer Society, brystkræft er den hyppigst diagnosticerede form for kræft hos kvinder i USA. Tidlig detektion kombineret med nyligt udviklede målrettede behandlinger har reduceret dødeligheden af ​​brystkræft i de sidste to årtier. Men brystkræft er stadig den næststørste årsag til kræft dødsfald hos kvinder i USA en. Størstedelen af ​​dødsfald blandt brystkræftpatienter skyldes tumorcellemetastase. Desværre er de fleste brystcancer er invasiv og ofte metastaserer til lymfeknuderne og efterfølgende til fjerne organer, herunder knogle, lunge, lever og hjerne. 1,2 Der er i øjeblikket ingen effektiv behandling for metastatisk brystcancer. Derfor udvikling af kemoterapeutiske og immunterapeutiske midler til at undertrykke metastatisk brystkræft er af stor betydning. Forskellige brystkræft spontan metastase musemodeller har bEen udviklet for at undersøge de molekylære mekanismer, der ligger brysttumor celle udvikling og metastaser og som skal anvendes som modeller til udvikling af terapeutiske midler. 1,3-5 Men de fleste af disse musemodeller er genetiske tumormodeller, at selv gode modeller for mekanistisk undersøgelser, er ikke egnede til testning terapeutiske midler eftersom metastase tager måneder at udvikle sig i disse genetiske modeller, hvilket kræver dyrt langvarig administration af anti-cancermidler. 6,7 Overvågning tumorprogression i levende mus er også teknisk udfordrende. Derimod kan et transplanteret brystkræft metastase-model har fordelene ved kortvarig tumorprogression og nem sporing af tumorprogression i levende mus. Den 4T1 ortotopisk brystcancer spontan metastase musemodel er sådan en transplanteret tumormodel. 8 I denne model er de brysttumorceller transplanteres ind i brystfedtpuden at etablere primære tumornoduler. Den primære tumorkan derefter fjernes kirurgisk som hos patienter human brystcancer. 4T1-tumorceller er yderst invasiv. 8,9, 3,10 Næsten alle tumorbærende mus udvikler metastase i 30 dage efter tumor transplantation ind i brystfedtpuden. Men 4T1 tumorer vokser aggressivt i de primære steder og tumor størrelser ofte overskrider de grænser, der er tilladt i de fleste dyr protokoller. På dette stadium, metastaserne er ofte mikrometastaser. Derfor er det vigtigt at fjerne primære tumorer for at tillade metastase fremskridt til afprøvning terapeutiske midler. Her rapporterer vi etableringen af en enkel og alligevel følsom procedure af primær tumor transplantation, kirurgisk fjernelse af primær tumor og bioluminescens imaging-baserede kvantificering af 4T1 tumorvækst og progression in vivo.

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an orthotopic mouse model of spontaneous breast cancer metastasis

Metastase er den primære årsag til dødelighed blandt brystkræftpatienter. Mekanismen bag cancercelle metastase, herunder brystcancer metastase, er stort set ukendt, og er fokus i cancerforskning. Forskellige brystkræft spontane metastaser musemodeller er blevet etableret. Her rapporterer vi en forenklet procedure for at etablere ortotopisk transplanteret brystkræft primær tumor og deraf spontan metastase, der efterligner menneskelig brystkræft metastaser. Kombineret med bioluminescens levende tumorafbildning, denne musemodel tillader tumorvækst og progression kinetik, der skal overvåges og kvantificeres. I denne model blev en lav dosis (1 x 10 4 celler) af 4T1-Luc brystcancerceller injiceret i BALB / c-mus brystfedtpude anvendelse af en tuberculinsprøjte. Mus blev injiceret med luciferin og afbildes på forskellige tidspunkter under anvendelse af et bioluminescerende billeddannende system. Når de primære tumorer voksede til størrelsen grænse i IACUC-godkendte protokol (ca.oximately 30 dage) blev musene bedøvet under konstant flow på 2% isofluran og oxygen. Tumorområdet blev steriliseret med 70% ethanol. Musen huden omkring tumoren blev udskåret for at blotlægge tumor, som blev fjernet med et par steril saks. Fjernelse af den primære tumor strækker overlevelse 4T-1 tumorbærende mus i en måned. Musene blev derefter gentagne gange afbildet for metastatisk tumor spredning til fjerne organer. Terapeutiske midler kan administreres for at undertrykke tumormetastase på dette punkt. Denne model er enkel og dog følsom kvantificere brystkræft cellevækst i det primære site og progression kinetik til fjerne organer, og dermed er en fremragende model til undersøgelse af brystkræft vækst og progression, og til afprøvning anti-metastase terapeutiske og immunterapeutiske midler in vivo .

Etablering af ortotopisk Breast Cancer Mouse Model 4T1 er en aggressiv brystcarcinoma-cellelinie. Injektion af så lidt som 1 x 10 4 celler i brystfedtpuden kan føre til etablering af en enkelt tumor knude i injektionsstedet (figur 2A). Derfor tumoren efterligner humane primære brystcarcinom. Næsten 100% mus udvikler den ortotopisk tumor. Tumorstørrelsen kan kvantificeres ved anvendelse af en digital skydelære eller ved udtag tumor...

Watch this Scientific Journal Video about An Orthotopic Mouse Model of Spontaneous Breast Cancer Metastasis at JoVE.com

An Orthotopic Mouse Model of Spontaneous Breast Cancer Metastasis

En orthotopisk brystcancer primær tumor model og kirurgisk fjernelse af den primære tumor for at forlænge muse liv til at generere spontan metastase er beskrevet. Den tumorvækst og progression overvåges og kvantificeres ved luciferase fluorescensafbildning.

En ortotopisk Mouse Model af Spontan Breast Cancer Metastase

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