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

Denne forskningen er finansiert av AACR Henry Shepard blærekreft Research Grant (09-60-30-LI).

Prosedyrer som involverer dyr fag har blitt godkjent av det institusjonelle Animal Care og bruk Committee (IACUC) ved University of California, San Francisco. 1. Celleforberedelsen <ol><li> Den menneskelige blærekreft celle linje KU-7-luc2-GFP (Caliper Life Sciences, Hopkinton, MA) dyrkes i RPMI-1640 media supplert med 10% fosterets bovint serum og 50 mikrogram / ml gentamicin. </li><li> Kulturer er opprettholdt ved 37 ° C, 5% CO 2 med media endres hver to til tre dager. </li>...

<ol>
	<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 presenterer en protokoll for etablering av en mus orthotopic xenograft blære tumor modell etterfulgt av intravesikal behandling av svulster med LNP formulert Sarna som er rettet mot arrangøren av P21 CIP1/WAF1 (P21) for transcriptional aktivering. 2, 3 Den resulterende svulster kan være bekreftet og overvåkes med Bioluminescent bildebehandling og blære ultralyd. Orthotopic modeller kan være overlegen intraperitoneal, subkutane eller intravenøse modeller ved å gi et miljø av urin og urot...

<table border="1"><tbody><tr><td> Navn på reagensen </td><td> Firma </td><td> Katalognummer </td><td> Kommentarer (valgfritt) </td></tr><tr><td> KU-7-Luc-GFP celler </td><td> Caliper </td><td> 128091 </td><td></td></tr><tr><td> thymic nakenmus (NU / NU) </td><td> Simonsen Laboratories </td><td> 6-7 uker gammel </td><td> Sim: (NCR) nu / nu fisol </td></tr><tr><td> Ultralyd enhet </td><td> Visualsonics </td><td> Vevo 770 </td><td> RMV-704 probe </td></tr><tr><td> Bioluminesens enhet </td><td> Caliper </td><td> IVIS Spectrum </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> Sølvnitrat </td><td> Sigma </td><td> S8157 </td><td></td></tr><tr><td> Hamilton sprøyte </td><td> Hamilton company </td><td> 80301 </td><td></td></tr><tr><td> LNP-Sarna </td><td> Alnylam Farmasi </td><td></td><td></td></tr></tbody></table> Tabell 1. Spesifikke reagenser og utstyr.

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

Vi presenterer en ny metode for behandling av blærekreft med intravesikalt levert liten aktiverende RNA (Sarna) i en orthotopic xenograft mus blære svulst modell. Musen modellen er etablert ved urethral kateterisering etter inhalert narkose. Kjemisk forbrenning blir så introdusert til blæren slimhinnene hjelp intravesikal sølvnitrat løsning å forstyrre blæren glycosaminoglycan laget og tillater celler å feste. Etter flere vask med sterilt vann, menneskelig blærekreft KU-7-luc2-GFP cellene er innpodet gjennom kateteret inn i blæren for å bo i 2 timer. Påfølgende vekst av blæretumorer bekreftes og overvåkes av in vivo blære ultralyd og Bioluminescent bildebehandling. Svulstene blir deretter behandlet intravesikalt med Sarna formulert i lipid nanopartikler (LNPs). Tumorvekst overvåkes med ultralyd og bioluminesens. Alle trinnene i denne fremgangsmåten er demonstrert i den medfølgende videoen.

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

Etablering en orthotopic blære tumor modell for å vurdere antitumor effekter av intravesikalt levert Sarna og overvåking tumorvekst ved ultralyd og Bioluminescent bildebehandling.

En Orthotopic Blære Tumor Model og bedømmelse av intravesikal Sarna Treatment

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

Research

JoVE Journal

Medicine

An Orthotopic Model of Murine Bladder Cancer

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and ...

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model

Kombinasjon Strålebehandling i en Orthotopic Mouse Brain Tumor Model

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

An Orthotopic Mouse Model of Anaplastic Thyroid Carcinoma

En orthotopic Mouse Modell av Anaplastisk thyroideakarsinom

Several types of animal models of human thyroid carcinomas have been established, including subcutaneous xenograft and orthotopic implantation of cancer ...

An Orthotopic Bladder Cancer Model for Gene Delivery Studies

En orthotopic Blærekreft Modell for Gene Levering Studies

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

En orthotopic Murine Model of Human Prostate Cancer Metastase

Our laboratory has developed a novel orthotopic implantation model of  human prostate cancer (PCa). As PCa death is not due to the primary tumor, but  ...

Minimally Invasive Establishment of Murine Orthotopic Bladder Xenografts

Minimal Invasive Etablering av Murint orthotopic Blære xenografts

Orthotopic bladder cancer xenografts are the gold standard to study molecular cellular manipulations and new therapeutic agents&#160;in vivo. Suitable ...

Evaluation of Tumor-infiltrating Leukocyte Subsets in a Subcutaneous Tumor Model

Evaluering av Tumor-infiltrere Leukocytt Delsett i en Subkutan Tumor Model

Specialized immune cells that infiltrate the tumor microenvironment regulate the growth and survival of neoplasia.&#160; Malignant cells must elude or ...

Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model

Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model

Vurdering av effektiviteten av<em&gt; H. pylori</em&gt; Protein HP-NAP som et terapeutisk verktøy for behandling av blærekreft i en ortotopiske Murine Model

Bladder cancer is one of the most common malignancies of the urogenital tract. Intravesical injection of Bacillus Calmette-Gu&#233;rin (BCG) is the gold ...

A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis

En &quot;Patient-Like&quot; ortotopiske syngeniske musemodell for leverkreft metastaser

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of ...