Name: experimental metastasis and ctl adoptive transfer immunotherapy mouse model
Uploaded: 2010-11-26T15:03:00
Description: Watch this Scientific Journal Video about Experimental Metastasis and CTL Adoptive Transfer Immunotherapy Mouse Model at JoVE.com

Finansierats med bidrag från National Institutes of Health (CA133085 till KL) och American Cancer Society (RSG-09-209-01-TBG till KL).

1. Experimentell Metastas musmodell <ol><li> Dagen innan tumörceller injektioner, utsäde en T75 kolv med upp till 1 x 10 7 CMS4-Met celler i 10 ml RPMI medium som innehåller 10% serum för att få snabbväxande tumörceller. Inkubera över natten vid 37 ° C. </li><li> På dagen för injektionen, ta bort mediet och skölj cellerna en gång med PBS, sedan skörda tumörceller med 0,05% trypsin-EDTA vid 37 ° C i 5 minuter. Stoppa reaktionen med 10 ml RPMI medium som innehåller 10% serum. Överför celler...

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	<li>Ryan, M. H., Bristol, J. A., McDuffie, E., Abrams, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regression+of+extensive+pulmonary+metastases+in+mice+by+adoptive+transfer+of+antigen-specific+CD8(+)+CTL+reactive+against+tumor+cells+expressing+a+naturally+occurring+rejection+epitope.">Regression of extensive pulmonary metastases in mice by adoptive transfer of antigen-specific CD8(+) CTL reactive against tumor cells expressing a naturally occurring rejection epitope.</a> J Immunol. 167 (8), 4286-4292 (2001).</li><li>Caldwell, S. A., Ryan, M. H., McDuffie, E., Abrams, S. 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E., Wunderlich, J. R., Yang, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adoptive+cell+transfer+therapy+following+non-myeloablative+but+lymphodepleting+chemotherapy+for+the+treatment+of+patients+with+refractory+metastatic+melanoma.">Adoptive cell transfer therapy following non-myeloablative but lymphodepleting chemotherapy for the treatment of patients with refractory metastatic melanoma.</a> J Clin Oncol. 23 (10), 2346-2357 (2005).</li><li>Srivastava, M. K., Sinha, P., Clements, V. K., Rodriguez, P., Ostrand-Rosenberg, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myeloid-derived+suppressor+cells+inhibit+T-cell+activation+by+depleting+cystine+and+cysteine.+Cancer+research.">Myeloid-derived suppressor cells inhibit T-cell activation by depleting cystine and cysteine. Cancer research.</a> 70 (1), 68-77 (2010).</li><li>Nagaraj, S., Gabrilovich, D. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+escape+mechanism+governed+by+myeloid-derived+suppressor+cells.">Tumor escape mechanism governed by myeloid-derived suppressor cells.</a> Cancer research. 68 (8), 2561-2563 (2008).</li><li>Nguyen, D. X., Bos, P. D., Massague, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metastasis:+from+dissemination+to+organ-specific+colonization.">Metastasis: from dissemination to organ-specific colonization.</a> Nature reviews. 9 (4), 274-284 (2009).</li><li>Heijstek, M. 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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IFN+Regulatory+Factor+8+Mediates+Apoptosis+in+Nonhemopoietic+Tumor+Cells+via+Regulation+of+Fas+Expression.">IFN Regulatory Factor 8 Mediates Apoptosis in Nonhemopoietic Tumor Cells via Regulation of Fas Expression.</a> J Immunol. 179 (7), 4775-4782 (2007).</li><li>Yang, D., Thangaraju, M., Greeneltch, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repression+of+IFN+regulatory+factor+8+by+DNA+methylation+is+a+molecular+determinant+of+apoptotic+resistance+and+metastatic+phenotype+in+metastatic+tumor+cells.">Repression of IFN regulatory factor 8 by DNA methylation is a molecular determinant of apoptotic resistance and metastatic phenotype in metastatic tumor cells.</a> Cancer research. 67 (7), 3301-3309 (2007).</li><li>Wexler, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accurate+identification+of+experimental+pulmonary+metastases.">Accurate identification of experimental pulmonary metastases.</a> Journal of the National Cancer Institute. 36 (4), 641-645 (1966).</li></ol>

Lösningar: Indien Bläck Solution (17): <ol><li> Häll 150 ml destillerat vatten till en 250 ml kolv. </li><li> Tillsätt 4 droppar ammoniumhydroxid med destillerat vatten. </li><li> Tillsätt 30 ml tusch lager (dvs Sanford Black Magic Vattentät tusch 4465 Item 44.011) till ammoniak och vatten-blandning. </li><li> Toppa med destillerat vatten till en volym av 200 ml. Lösningen är färdig för injektion. </li></ol> Fekete lösning (17): Fekete lösning används för att bleka tusch-uppblåsta tumör-bärande lungorna för att skilja vitt tumörnoduli från den svarta bakgrunden av normal vävnad. <ol><li> Tillsätt 350 ml 95% EtOH till 1L glasflaska. </li><li> Tillsätt 150ml destillerat vatten </li><li> Lägg 50ml formaldehyd </li><li> Lägg 25ml koncentrerad sura syra </li></ol>

experimental metastasis and ctl adoptive transfer immunotherapy mouse model

Experimentell metastaser musmodell är en enkel och ändå fysiologiskt relevanta metastaser modell. Tumörcellerna injiceras intravenöst (iv) i vener mus svans och kolonisera i lungorna, vilket, som liknar de sista stegen i tumörcellen spontana metastaser: överlevnad i cirkulationen, extravasering och kolonisation i distala organ. Från en terapeutisk synvinkel är den experimentella metastas modellen enklaste och ideala modellen eftersom målet av terapier är ofta slutpunkten för metastas: etablerade metastaserande tumör i distala orgel. I denna modell är tumörcellerna injiceras iv i mus svans vener och tillåtas att kolonisera och växa i lungorna. Tumör-specifik CTL sedan injiceras iv i metastaser bärande mus. Antalet och storleken på lungmetastaser kan styras av antalet tumörceller som ska injiceras och tidpunkten för tumörtillväxt. Därför är olika stadier av metastaser, från minimal metastaser till omfattande metastasering, kan modelleras. Lungmetastaser analyseras genom inflation med bläck, vilket gör det lättare att visuell observation och kvantifiering.

Watch this Scientific Journal Video about Experimental Metastasis and CTL Adoptive Transfer Immunotherapy Mouse Model at JoVE.com

Experimental Metastasis and CTL Adoptive Transfer Immunotherapy Mouse Model

En experimentell lungmetastaser och CTL immunterapi musmodell för analys av tumörceller-T-cell interaktion<em&gt; In vivo</em&gt;.

Experimentell Metastas och CTL adoptiv transfer Immunterapi musmodell

Experimental metastasis mouse model is a simple and yet physiologically relevant metastasis model. The tumor cells are injected intravenously (i.v) into ...

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