Name: combination radiotherapy in an orthotopic mouse brain tumor model
Uploaded: 2012-03-06T12:00:00
Description: Watch this Scientific Journal Video about Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model at JoVE.com

Denne forskning er støttet med midler fra Egne program NIH.

I. Cellepræparation <ol><li> Cellerne dyrkes til sammenflydning i DMEM med 10% FBS omkring en 80% konfluente 225 cm3 kolbe for hver 5 mus, der implanteres. </li><li> Aspireres mediet ud af cellerne, vaskes med 10 ml PBS uden calcium eller magnesium. Aspirer PBS off-celler. </li><li> Trypsinisér cellerne med 3 ml trypsin, vente 10-15 minutter, og neutralisere trypsin med 15 ml medium. </li><li> Pipette alle celler og medier i en 50 ml konisk. To store kolber passer ind i en 50 ml konisk. </li><li> Spinde c...

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	<li>Bleau, A. M., Holland, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trapping+the+mouse+genome+to+hunt+human+alternations.">Trapping the mouse genome to hunt human alternations.</a> Proc. Nat. Acad. Sci. U.S.A. 104, 7737-7738 (2007).</li><li>McGrady, B. J., McCormick, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+murine+model+of+intracranial+invasion:+morphological+observations+on+central+nervous+system+invasion+by+melanoma+cells.">A murine model of intracranial invasion: morphological observations on central nervous system invasion by melanoma cells.</a> Clin. Exp. Metastasis. 10, 387-393 (1992).</li><li>Fei, X., Zhang, Q., Dong, J., Diao, y., Wang, Z., Li, R., Wu, Z., Wang, A., Lan, Q., Zhang, S., Huang, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+clinically+relevant+orthotopic+xenograft+mouse+model+of+metastatic+lung+cancer+and+glioblastoma+through+surgical+tumor+tissues+injection+with+trocar.">Development of clinically relevant orthotopic xenograft mouse model of metastatic lung cancer and glioblastoma through surgical tumor tissues injection with trocar.</a> Journal of Experimental &#38; Clinical Cancer Research. 29, 84-84 (2010).</li><li>Fujita, M., Zhu, X., Sasaki, K., Ueda, R., Low, K., Pollack, I., Okada, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+STAT3+Promotes+the+Efficacy+of+Adoptive+Transfer+Therapy+Using+Type-1+CTLs+by+Modulation+of+the+Immunological+Microenvironment+in+a+Murine+Intracranial+Glioma.">Inhibition of STAT3 Promotes the Efficacy of Adoptive Transfer Therapy Using Type-1 CTLs by Modulation of the Immunological Microenvironment in a Murine Intracranial Glioma.</a> J. Immunol. 180, 2089-2098 (2008).</li><li>Candolfi, M., Curtin, J. F., Nichols, W. S., Muhammad, A. K. M. .. . G., King, G., Pluhar, G. D., McNiel, G. E., Ohlfest, E. A., Freese, J. R., Moore, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracranial+glioblastoma+models+in+preclinical+neuro-oncology:+neuropathlogical+characterization+and+tumor+progression.">Intracranial glioblastoma models in preclinical neuro-oncology: neuropathlogical characterization and tumor progression.</a> J. Neurooncol. 85, 133-148 (2007).</li><li>Kaye, A., Morstyn, G., Gardner, I., Pyke, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+xenograft+glioma+model+in+mouse+brain.">Development of a xenograft glioma model in mouse brain.</a> Cancer Research. 46, 1367-1373 (1986).</li><li>Zhao, Y., Xiao, A., diPierro, C., Carpenter, J., Abdel-Fattah, R., Redpath, G., Lopez, M., Hussaini, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+Extensive+Invasive+Intracranial+Human+Glioblastoma+Xenograft+Model.">An Extensive Invasive Intracranial Human Glioblastoma Xenograft Model.</a> American Journal of Pathology. 176, 3032-3049 (2010).</li><li>Learn, C., Grossi, P., Schmittling, R., Xie, W., Mitchell, D., Karikari, I., Wei, Z., Dressman, H., Sampson, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+Analysis+of+Intracranial+Tumors+in+a+Murine+Model+of+Glioma+Demonstrate+a+Shift+in+Gene+Expression+in+Response+to+Host+Immunity.">Genetic Analysis of Intracranial Tumors in a Murine Model of Glioma Demonstrate a Shift in Gene Expression in Response to Host Immunity.</a> J. Neuroimmunol. 182, 63-72 (2007).</li><li>Martens, T., Laabs, Y., Günther, H., Kemming, D., Zhu, Z., Witte, L., Hagel, C., Westphal, M., Lamszus, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+Glioblastoma+Growth+in+a+Highly+Invasive+Nude+Mouse+Model+Can+Be+Achieved+by+Targeting+Epidermal+Growth+Factor+Receptor+but+not+Vascular+Endothelial+Growth+Factor+Receptor-2.">Inhibition of Glioblastoma Growth in a Highly Invasive Nude Mouse Model Can Be Achieved by Targeting Epidermal Growth Factor Receptor but not Vascular Endothelial Growth Factor Receptor-2.</a> Clin. Cancer Res. 14, 5447-5458 (2008).</li><li>Purow, B., Schiff, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+the+genetics+of+glioblastoma:+are+we+reaching+critical+mass.">Advances in the genetics of glioblastoma: are we reaching critical mass.</a> Nat. Rev. Neurol. 5, 419-426 (2009).</li></ol>

Maligne gliomer, såsom glioblastoma multiforme, repræsenterer de mest almindelige primære hjernetumorer og har en trist prognose 4. Overlevelse af patienter, der lider af GBM er forblevet stort set uændret i løbet af de sidste ti år (dvs. 9-12 måneder efter diagnose) på trods af fremskridt inden for kirurgi, strålebehandling og kemoterapi 5. Trækkene ved en passende model til undersøgelse af glioma behandling bør indeholde en reproducerbar placering af tumor og vækst af celler som et r...

<table border="1"><tbody><tr><td> Lab Standard stereotaktiske Instrutment </td><td> Stoelting mus og rotte Adaptor </td></tr><tr><td> Betadine </td><td> 100% EtOH </td></tr><tr><td> 70% EtOH </td><td> 50% acetone </td></tr><tr><td> 6 &quot;Q-tip </td><td> Spritservietter </td></tr><tr><td> Hamilton sprøjte 80.008 </td><td> Ethicon 5-0 PD </td></tr><tr><td> 1701 SN 31-33G / 1 &quot;/ PT3 nål </td><td> 1 ml injektionssprøjter </td></tr><tr><td> 25G kanyler </td><td> Varme pad </td></tr><tr><td> Kunstig Tears Salve </td><td> Meget fin tippet permanent markør </td></tr></tbody></table>

combination radiotherapy in an orthotopic mouse brain tumor model

Glioblastoma multiforme (GBM) er den mest almindelige og aggressive voksne primære hjernetumorer 1. I de senere år har der været betydelige fremskridt i forståelsen af mekanikken af tumor invasion, og direkte intracerebral podning af tumor giver mulighed for at observere den invasive proces i et fysiologisk egnet miljø 2. For så vidt angår humane hjernetumorer vedkommende er orthotopisk model i øjeblikket er tilgængelige enten ved stereotaktisk injektion af cellesuspensioner eller implantering af et massivt stykke tumor via en kompliceret kraniotomi procedure 3. I vores teknik vi høstes celler fra vævskultur for at skabe en cellesuspension, der anvendes til at implantere direkte ind i hjernen. Varigheden af ​​operationen er cirka 30 minutter, og da musen skal være i en konstant kirurgisk plan, er en injicerbar anæstetikum. Musen anbringes i en stereotaktisk jig ved Stoetling (figur 1). Aftis det kirurgiske område renses og fremstilles, er et snit, og bregma er placeret til at bestemme placeringen af ​​kraniotomi. Placeringen af ​​kraniotomi er 2 mm til højre og 1 mm rostralt for bregma. Dybden er 3 mm fra overfladen af ​​skallen, og cellerne injiceres ved en hastighed på 2 pi hvert 2. minut. Huden sutureres med 5-0 PDS, og musen bliver lov til at vågne op på en varmepude. Fra vores erfaring, afhængigt af den cellelinie kan behandlingen foregå fra 7-10 dage efter kirurgi. Afgivelsesenheden er afhængig af lægemiddelsammensætningen. For strålebehandling musene er bedøvet, og sat ind i en skræddersyet jig. Bly omfatter musens krop og udsætter kun hjernen på musen. Studiet af tumorigenese og evaluering af nye terapier for GBM kræver nøjagtige og reproducerbare hjernesvulst dyremodeller. Således har vi anvende denne orthotopisk hjerne model til at studere interaktionen af ​​mikromiljøet af hjernen og tumoren, til at teste effectivenESS af forskellige terapeutiske midler med og uden stråling.

Watch this Scientific Journal Video about Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model at JoVE.com

Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model

Formålet med denne artikel er at beskrive brugen af ​​en orthotopisk glioblastom model for chemoradiation undersøgelser. Denne artikel vil gå gennem celle behandling, implantering, samt strålebehandling af musen ved hjælp af en intrakraniel model.

Kombinationen Radioterapi i en orthotopisk musehjerne tumormodel

Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the ...

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