Name: an orthotopic bladder tumor model and the evaluation of intravesical sarna treatment
Uploaded: 2012-07-28T14:00:00
Description: Watch this Scientific Journal Video about An Orthotopic Bladder Tumor Model and the Evaluation of Intravesical saRNA Treatment at JoVE.com

Denna forskning finansieras av AACR Henry Shepard urinblåsan Cancer Research Grant (09-60-30-LI).

Förfaranden som inbegriper djurförsök har godkänts av Institutional Animal Care and Use Committee (lACUC) vid University of California, San Francisco. 1. Cellberedning <ol><li> Den humana cancer i urinblåsan cellinje KU-7-luc2-GFP (Caliper Life Sciences, Hopkinton, MA) odlas i RPMI-1640 medium kompletterat med 10% fetalt bovint serum och 50 | ig / ml gentamicin. </li><li> Kulturerna upprätthölls vid 37 ° C, 5% CO2 med mediet ändras var två till tre dagar. </li><li> Cel...

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	<li>Semple, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rational+design+of+cationic+lipids+for+siRNA+delivery.">Rational design of cationic lipids for siRNA delivery.</a> Nature. 28, 172-176 (2010).</li><li>Li, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+dsRNAs+induce+transcriptional+activation+in+human+cells.">Small dsRNAs induce transcriptional activation in human cells.</a> Proceedings of the National Academy of Sciences of the United States of America. 103, 17337-17342 (2006).</li><li>Chen, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antitumor+effect+of+dsRNA-induced+p21(WAF1/CIP1)+gene+activation+in+human+bladder+cancer+cells.">Antitumor effect of dsRNA-induced p21(WAF1/CIP1) gene activation in human bladder cancer cells.</a> Molecular cancer therapeutics. 7, 698-703 (2008).</li><li>Chancellor, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preliminary+results+of+myoblast+injection+into+the+urethra+and+bladder+wall:+a+possible+method+for+the+treatment+of+stress+urinary+incontinence+and+impaired+detrusor+contractility.">Preliminary results of myoblast injection into the urethra and bladder wall: a possible method for the treatment of stress urinary incontinence and impaired detrusor contractility.</a> Neurourology and urodynamics. 19, 279-287 (2000).</li><li>Hadaschik, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+validated+mouse+model+for+orthotopic+bladder+cancer+using+transurethral+tumour+inoculation+and+bioluminescence+imaging.">A validated mouse model for orthotopic bladder cancer using transurethral tumour inoculation and bioluminescence imaging.</a> BJU international. 100, 1377-1384 (2007).</li><li>Fu, C., Apelo, C. A., Torres, B., Thai, K. H., Hsieh, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+Bladder+Wall+Injection.">Mouse Bladder Wall Injection.</a> J. Vis. Exp. (53), e2523 (2011).</li><li>Dobek, G. L., Godbey, W. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+Orthotopic+Model+of+Murine+Bladder.">An Orthotopic Model of Murine Bladder.</a> Cancer. J. Vis. Exp. (48), 2535 (2011).</li><li>Dinney, C. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+characterization+of+metastatic+variants+from+human+transitional+cell+carcinoma+passaged+by+orthotopic+implantation+in+athymic+nude+mice.">Isolation and characterization of metastatic variants from human transitional cell carcinoma passaged by orthotopic implantation in athymic nude mice.</a> The Journal of urology. 154, 1532-1538 (1995).</li></ol>

Vi presenterar ett protokoll för upprättandet av en mus ortotopisk xenograft urinblåsan tumörmodellen följt av intravesikal behandling av tumörer med LNP formulerade Sarna som riktar initiativtagaren till p21 CIP1/WAF1 (p21) för transkriptionsaktivering. 2, 3 De resulterande tumörer kan vara bekräftas och övervakas med självlysande avbildning och urinblåsa ultraljud. Ortotopisk modell kan vara överlägsen intraperitoneal, subkutan eller intravenösa modeller genom att tillhandahålla e...

<table border="1"><tbody><tr><td> Namnet på reagens </td><td> Företaget </td><td> Katalognummer </td><td> Kommentarer (valfri) </td></tr><tr><td> KU-7-Luc-GFP-celler </td><td> Bromsok </td><td> 128.091 </td><td></td></tr><tr><td> tymiska nakna möss (nu / nu) </td><td> Simonsen Laboratories </td><td> 6-7 veckor gamla </td><td> Sim: (NCR) nu / nu fisol </td></tr><tr><td> Ultraljudsenheten </td><td> Visualsonics </td><td> VEVO 770 </td><td> RMV-704 givare </td></tr><tr><td> Bioluminiscens enhet </td><td> Bromsok </td><td> IVIS spektrum </td><td></td></tr><tr><td> Exel safelet kateter </td><td> Fisher </td><td> 14-841-21 </td><td> 24G x ¾ &quot; </td></tr><tr><td> Silvernitrat </td><td> Sigma </td><td> S8157 </td><td></td></tr><tr><td> Hamilton-spruta </td><td> Hamilton company </td><td> 80.301 </td><td></td></tr><tr><td> LNP-Särna </td><td> Alnylam Läkemedel </td><td></td><td></td></tr></tbody></table> Tabell 1. Specifika reagens och utrustning.

an orthotopic bladder tumor model and the evaluation of intravesical sarna treatment

Vi presenterar ett nytt förfarande för behandling av blåscancer med intravesikalt levereras liten aktiverande RNA (Sarna) i en ortotopisk xenotransplantat mus blåsan tumörmodell. Den musmodell fastställs genom urinröret kateterisering i inhalerade narkos. Kemisk brännskada införs sedan till blåsan slemhinna under användning av intravesikalt silvernitratlösning att störa skiktet blåsan glykosaminoglykan och tillåter celler att fästa. Efter flera tvättar med sterilt vatten, blåscancer hos människa KU-7-luc2-GFP celler instilleras genom katetern in i blåsan för att verka i 2 timmar. Efterföljande tillväxt av tumörer i urinblåsan bekräftas och övervakas av in vivo urinblåsan ultraljud och självlysande bilder. Tumörerna behandlas sedan intravesikalt med Sarna formulerat i lipid nanopartiklar (LNPs). Tumörtillväxten övervakas med ultraljud och bioluminiscens. Alla steg i detta förfarande är visat på bifogade videon.

Watch this Scientific Journal Video about An Orthotopic Bladder Tumor Model and the Evaluation of Intravesical saRNA Treatment at JoVE.com

An Orthotopic Bladder Tumor Model and the Evaluation of Intravesical saRNA Treatment

Upprätta en ortotopisk modell blåsa tumören för att utvärdera antitumöreffekter av intravesikalt levererat Särna och övervakning tumörtillväxt genom ultraljud och självlysande bilder.

En ortotopisk blåstumör modell och utvärdering av Intravesikal Sarna behandling

We present a novel method for treating bladder cancer with intravesically delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder ...

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