Name: generation of a liver orthotopic human uveal melanoma xenograft platform in immunodeficient mice
Uploaded: 2019-11-06T13:00:00
Description: Watch this Scientific Journal Video about Generation of a Liver Orthotopic Human Uveal Melanoma Xenograft Platform in Immunodeficient Mice Video at JoVE.com

Vi er takknemlige for M. Ohara, K. Saito, og M. Terai, for å ha gjennomgått manuskriptet. Forfatterne erkjenner kritisk vurdering for redaksjonelle og engelsk bistand av dette manuskriptet av Dr. R. Sato på Fox Chase Cancer Center. Arbeidet beskrevet her ble støttet av Bonnie Kroll Research Fund, The Merk Weinzierl Research Fund, Eye melanom Research Fund ved Thomas Jefferson University, The Osaka Community Foundation, og JSP KAKENHI Grant Number JP 18K15596 i Osaka City University. Studier i Dr. A. Aplin laboratorium ble støttet av NIH Grant R01 GM067893. Dette prosjektet ble også finansiert av en Dean ' s bearbeidet Science Award, en Thomas Jefferson University programmatisk Initiative Award.

Pasienter som deltar i studien bør gi skriftlig samtykke som tillater bruk av kasserte kirurgiske prøver for forskningsformål og genetiske studier, ifølge en institusjonell gjennomgang Board-godkjent protokoll. Denne protokollen ble utført i nøye samsvar med anbefalingene i guide for omsorg og bruk av laboratorium dyr av National Institutes of Health og godkjent av institusjonelle Animal Care og use Committee (IACUC).
1. innsamling av frisk pasient-avledet tumor vev
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<ol>
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S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Survival+of+patients+with+metastases+from+uveal+melanoma.">Survival of patients with metastases from uveal melanoma.</a> Ophthalmology. 98 (3), 383-389 (1991).</li><li>Diener-West, M., et al. <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+metastatic+disease+after+enrollment+in+the+COMS+trials+for+treatment+of+choroidal+melanoma:+Collaborative+Ocular+Melanoma+Study+Group+Report+No.+26.">Development of metastatic disease after enrollment in the COMS trials for treatment of choroidal melanoma: Collaborative Ocular Melanoma Study Group Report No. 26.</a> Archives of Ophthalmology. 123 (12), 1639-1643 (2005).</li><li>Collaborative Ocular Melanoma Study Group. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+metastatic+disease+status+at+death+in+435+patients+with+large+choroidal+melanoma+in+the+Collaborative+Ocular+Melanoma+Study+(COMS):+COMS+report+no.+15.">Assessment of metastatic disease status at death in 435 patients with large choroidal melanoma in the Collaborative Ocular Melanoma Study (COMS): COMS report no. 15.</a> Archives of Ophthalmology. 119 (5), 670-676 (2001).</li><li>Hidalgo, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patient-derived+xenograft+models:+an+emerging+platform+for+translational+cancer+research.">Patient-derived xenograft models: an emerging platform for translational cancer research.</a> Cancer Discovery. 4 (9), 998-1013 (2014).</li><li>Kim, M. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+orthotopic+and+heterotopic+human+pancreatic+cancer+xenografts+in+immunodeficient+mice.">Generation of orthotopic and heterotopic human pancreatic cancer xenografts in immunodeficient mice.</a> Nature Protocols. 4 (11), 1670-1680 (2009).</li><li>N&#233;mati, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+and+characterization+of+a+panel+of+human+uveal+melanoma+xenografts+derived+from+primary+and/or+metastatic+tumors.">Establishment and characterization of a panel of human uveal melanoma xenografts derived from primary and/or metastatic tumors.</a> Clinical Cancer Research. 16 (8), 2352-2362 (2010).</li><li>Wilmanns, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+Doxorubicin+sensitivity+and+level+of+p-glycoprotein+expression+in+human+colon-carcinoma+cells+by+ectopic+and+orthotopic+environments+in+nude-mice.">Modulation of Doxorubicin sensitivity and level of p-glycoprotein expression in human colon-carcinoma cells by ectopic and orthotopic environments in nude-mice.</a> International Journal of Oncology. 3 (3), 413-422 (1993).</li><li>Kang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proliferation+of+human+lung+cancer+in+an+orthotopic+transplantation+mouse+model.">Proliferation of human lung cancer in an orthotopic transplantation mouse model.</a> Experimental and Therapeutic. 1 (3), 471-475 (2010).</li><li>Fichtner, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+of+patient-derived+non-small+cell+lung+cancer+xenografts+as+models+for+the+identification+of+predictive+biomarkers.">Establishment of patient-derived non-small cell lung cancer xenografts as models for the identification of predictive biomarkers.</a> Clinical Cancer Research. 14 (20), 6456-6468 (2008).</li><li>Marangoni, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+model+of+patient+tumor-derived+breast+cancer+xenografts+for+preclinical+assays.">A new model of patient tumor-derived breast cancer xenografts for preclinical assays.</a> Clinical Cancer Research. 13 (13), 3989-3998 (2007).</li><li>Bergamaschi, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+profiling+and+characterization+of+luminal-like+and+basal-like+in+vivo+breast+cancer+xenograft+models.">Molecular profiling and characterization of luminal-like and basal-like in vivo breast cancer xenograft models.</a> Molecular Oncology. 3 (5-6), 469-482 (2009).</li><li>Ho, K. S., Poon, P. C., Owen, S. C., Shoichet, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+vessel+hyperpermeability+and+pathophysiology+in+human+tumour+xenograft+models+of+breast+cancer:+a+comparison+of+ectopic+and+orthotopic+tumours.">Blood vessel hyperpermeability and pathophysiology in human tumour xenograft models of breast cancer: a comparison of ectopic and orthotopic tumours.</a> BMC Cancer. 12, 579 (2012).</li><li>Hoffman, R. 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Y., Besterman, J. M., Monosov, A., Hoffman, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+of+human+metastatic+colon+cancer+in+nude+mice+orthotopically+constructed+by+using+histologically+intact+patient+specimens.">Models of human metastatic colon cancer in nude mice orthotopically constructed by using histologically intact patient specimens.</a> Proceedings of the National Academy of Sciences of the United States of America. 88 (20), 9345-9349 (1991).</li><li>Rashidi, B., et al. <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+mouse+model+of+remetastasis+of+human+colon+cancer+liver+metastasis.">An orthotopic mouse model of remetastasis of human colon cancer liver metastasis.</a> Clinical Cancer Research. 6 (6), 2556-2561 (2000).</li><li>Fan, Z. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Real-time+monitoring+of+rare+circulating+hepatocellular+carcinoma+cells+in+an+orthotopic+model+by+in+vivo+flow+cytometry+assesses+resection+on+metastasis.">Real-time monitoring of rare circulating hepatocellular carcinoma cells in an orthotopic model by in vivo flow cytometry assesses resection on metastasis.</a> Cancer Research. 72 (10), 2683-2691 (2012).</li><li>Jacob, D., Davis, J., Fang, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Xenograftictumor+modelsinmiceforcancer+research,+atechnical+review.">Xenograftictumor modelsinmiceforcancer research, atechnical review.</a> Gene Therapy and Molecular Biology. 8, 213-219 (2004).</li><li>Ahmed, S. 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De nåværende orthotopic xenograft modellene er arbeidskrevende, tidkrevende og kostbart å skape. Orthotopic tumor xenograft mus modeller for leverkreft ble etablert mer enn to ti år siden19,20,21. Imidlertid er denne teknikken komplisert og krever bruk av spesialutstyr, for eksempel en mikro-nål holderen og 6-0 til 8-0 fine sting under et mikroskop. Tumor og normalt lever vev må sys opp nøye slik at Sutur ikke skader den ...

Uveal melanom er den vanligste intraokulært ondartet svulst blant voksne i den vestlige verden. I løpet av de siste 50 årene har forekomsten av uveal melanom (5,1 tilfeller per million) forblitt stabil i USA1,2. Uveal melanom oppstår fra melanocytter i Iris, ciliary kroppen, eller akkord, og det er en ekstremt dødelig sykdom når den utvikler metastasering. Dødeligheten av pasienter med uveal melanom metastasering var 80% ved 1 år og 92% ved 2 år etter første diagnostisering av metastaser. Tiden mellom diagnostisering av metastaser og død er vanligvis kort, mindre enn 6 måneder, uten hensyn til terapi3,4. Kreften sprer seg gjennom blodet og har en tendens til å nesten kun metastase til leveren (89-93%)4,5. En effektiv musemodell er et presserende behov for videre etterforskning av leveren-metastatisk uveal melanom. For translational forskning, det er et klart behov for å generere en lever-lokaliserte metastatisk uveal melanom musemodell.
En pasient-avledet tumor xenograft (PDX) musemodeller forventes å gi individualisert medisin strategier. Disse modellene kan være prediktiv av kliniske utfall, være nyttig for prekliniske narkotika evaluering, og brukes til biologiske studier av svulster6. Representative PDX-modeller er jektopicheski tumor-implantert xenograft mus, som har tumor på subkutan nettsteder. De fleste forskere kan gjøre kirurgi for subkutan implantation uten spesiell praksis7,8. De kan også overvåke subkutan svulster lett. Selv om subkutan PDX-modeller ble populære i forsknings fasen, har de noen hindringer i bevegelse til praktisk bruk. Subkutan implantation styrker pasient-avledede svulster å engraft på en annen mikromiljøet fra tumor opprinnelse, slik at det fører til engraftment svikt og langsom tumor vekst 9,10,11, 12,13,14. Orthotopic engraftment kan være en mer ideell og rasjonell tilnærming for en PDX modell fordi den bruker samme organ som den opprinnelige tumor15,16.
Nylig utviklet vi protokoller for kirurgiske orthotopic implantation teknikker av pasient-avledet lever-metastatisk uveal melanom svulster og nål injeksjon teknikker med en kultivert menneskelig lever-metastatisk uveal melanom cellelinje i NOD. CG-PrkdcscidIl2rgtm1Wjl/SzJ (nsg) mus17,18. Protokollene resulterer i gjennomgående høye tekniske suksessrater. Vi har også etablert CT-skanning teknikker som er nyttige for å oppdage interiør leverskader svulster, og vi utviklet ny implantation av embryo svulster i PDX plattform. Vi fant at uveal melanom tumor xenograft modeller opprettholde egenskapene til den opprinnelige pasienten leveren svulst, inkludert deres histopathological og molekylære funksjoner. Sammen, disse teknikkene gir en bedre plattform for leveren orthotopic tumor modeller for uveal melanom i translational forskning.

<table>
	<tbody>
		<tr>
			<td>Materials, tissues and animals</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>CO2 tank</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cryomedium</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Exitron nano 12000 (Alkaline earth metal-based nanoparticle contrast agent)</td>
			<td>Miltenyl Biotec</td>
			<td>130-095-700</td>
			<td></td>
		</tr>
		<tr>
			<td>HBSS 1X, with calcium &#38; magnesium</td>
			<td>Corning</td>
			<td>21-020-CM</td>
			<td></td>
		</tr>
		<tr>
			<td>Human liver metastatic uveal melanoma cell line</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Human uveal melanoma tissue in the liver</td>
			<td></td>
			<td></td>
			<td>All tissue handling should be done in a Biosafety Level 2 hood. Be careful when working with human tissue; always use gloves and avoid direct skin contact. Assume patients may have been infected with HIV or other highly transmissible organisms. Do not process samples known to carry infections.</td>
		</tr>
		<tr>
			<td>Iodine</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Purdue Products</td>
			<td>67618-150-17</td>
			<td></td>
		</tr>
		<tr>
			<td>Isopropanol</td>
			<td>Fisher scientific</td>
			<td>A416-1</td>
			<td>Avoid direct contact to skin and eye and inhalation of anesthetic agent.</td>
		</tr>
		<tr>
			<td>Liquid nitrogen</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Matrigel HC</td>
			<td>BD</td>
			<td>354248</td>
			<td></td>
		</tr>
		<tr>
			<td>NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice</td>
			<td>Jackson Lab</td>
			<td>5557</td>
			<td>4 to 8 weeks old</td>
		</tr>
		<tr>
			<td>PBS 1X, without calcium and magnesium</td>
			<td>Corning</td>
			<td>21-031-CM</td>
			<td></td>
		</tr>
		<tr>
			<td>RPMI 1640</td>
			<td>Corning</td>
			<td>10-013-CV</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile alcohol prep pad (70% isopropyl alcohol)</td>
			<td>Nice-Pak products</td>
			<td>B603</td>
			<td></td>
		</tr>
		<tr>
			<td>4% paraformaldehyde phosphate buffer solution</td>
			<td>Wako</td>
			<td>163-20145</td>
			<td></td>
		</tr>
		<tr>
			<td>70% Ethyl alcohol solution</td>
			<td>Fisher Scientific</td>
			<td>04-355-122</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Equipments</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Absorbable hemostat</td>
			<td>Johnson and Johnson</td>
			<td>63713-0019-61</td>
			<td></td>
		</tr>
		<tr>
			<td>Autoclave</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Body weight measure</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cautery</td>
			<td>Bovie Medical</td>
			<td>MC-23009</td>
			<td></td>
		</tr>
		<tr>
			<td>Cell counter</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuzer</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton swab</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cryo freezing container</td>
			<td>NALGENE</td>
			<td>5100-0001</td>
			<td></td>
		</tr>
		<tr>
			<td>Cryotube</td>
			<td>SARSTEDT</td>
			<td>72.379</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved scissors</td>
			<td>World Precision Instruments</td>
			<td>503247</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved ultrafine forceps</td>
			<td>World Precision Instruments</td>
			<td>501302</td>
			<td></td>
		</tr>
		<tr>
			<td>Fabric sheet</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Freezer</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>F/AIR Filter Canister</td>
			<td>Harvard Apparatus</td>
			<td>600979</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane vaporizer</td>
			<td>Artisan Scientific</td>
			<td>66317-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Liquid nitrogen</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Liquid nitrogen jar</td>
			<td>Thermo Fisher Scientific</td>
			<td>2123</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-CT scan</td>
			<td>Siemens</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>World Precision Instruments</td>
			<td>501246</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri dishes</td>
			<td>Fisher Scientific</td>
			<td>FB0875713</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipette</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Spray bottle</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile hood</td>
			<td></td>
			<td></td>
			<td>Biosafety level 2 cabinet</td>
		</tr>
		<tr>
			<td>Sterile No.11 scalpel</td>
			<td>AD Surgical</td>
			<td>A300-11-0</td>
			<td></td>
		</tr>
		<tr>
			<td>Straight forceps</td>
			<td>World Precision Instruments</td>
			<td>14226</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical drape</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Tail vein restrainer</td>
			<td>Braintree Scientific</td>
			<td>TV-150-STD</td>
			<td></td>
		</tr>
		<tr>
			<td>Water bath</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1 ml TB syringe with 27-gauge needle</td>
			<td>BD</td>
			<td>309623</td>
			<td></td>
		</tr>
		<tr>
			<td>1.7 ml tube</td>
			<td>Bioexpress</td>
			<td>C-3260-1</td>
			<td></td>
		</tr>
		<tr>
			<td>5-0 PDO Suture</td>
			<td>AD Surgical</td>
			<td>S-D518R13</td>
			<td></td>
		</tr>
		<tr>
			<td>15 mL conical tubes</td>
			<td>AZER SCIENTIFIC</td>
			<td>ES-9152N</td>
			<td></td>
		</tr>
		<tr>
			<td>27-gauge needle</td>
			<td>BD</td>
			<td>780301</td>
			<td></td>
		</tr>
		<tr>
			<td>27-gauge needle</td>
			<td>Hamilton</td>
			<td>7803-01</td>
			<td></td>
		</tr>
		<tr>
			<td>50 mL conical tubes</td>
			<td>AZER SCIENTIFIC</td>
			<td>ES-9502N</td>
			<td></td>
		</tr>
		<tr>
			<td>50 &#181;l micro syringe</td>
			<td>BD</td>
			<td>80630</td>
			<td></td>
		</tr>
		<tr>
			<td>50 &#181;l micro syringe</td>
			<td>Hamilton</td>
			<td>7655-01</td>
			<td></td>
		</tr>
		<tr>
			<td>100 mL container</td>
			<td>Fisher Scientific</td>
			<td>12594997</td>
			<td></td>
		</tr>
		<tr>
			<td>200&#956;l tip</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

generation of a liver orthotopic human uveal melanoma xenograft platform in immunodeficient mice

I de siste ti årene, subkutant implantert pasient-avledet xenograft svulster eller kultivert menneskelige cellelinjer har blitt stadig mer anerkjent som mer representative modeller for å studere menneskelig kreft i immunodeficient mus enn tradisjonelle etablerte menneskelige celle linjer in vitro. Nylig, orthotopically implantert pasient-avledet tumor xenograft (PDX) modeller i mus har blitt utviklet for å bedre gjenskape funksjoner av pasient svulster. En lever orthotopic xenograft musen modell forventes å bli en nyttig kreften forskning plattform, skaffer innblikk i svulst Biology og bedøve terapi. Imidlertid er leveren orthotopic tumor implantation generelt komplisert. Her beskriver vi våre protokoller for orthotopic implantation av pasient-avledet lever-metastatisk uveal melanom svulster. Vi kultivert menneskelig lever metastatisk uveal melanom cellelinjer i immunodeficient mus. Protokollene kan resultere i gjennomgående høy teknisk suksess priser ved hjelp av enten en kirurgisk orthotopic implantation teknikk med biter av pasient-avledet uveal melanom tumor eller en nål injeksjon teknikk med kultivert menneskelig cellelinje. Vi beskriver også protokoller for CT skanning å oppdage interiør leversvulster og for re-implantation teknikker bruker embryo svulster å oppnå re-engraftment. Sammen, disse protokollene gir en bedre plattform for leveren orthotopic tumor mus modeller av leveren metastatisk uveal melanom i translational forskning.

Kirurgisk orthotopic implantation bruker leveren Pocket metoden kan transplantasjon menneskelig leveren metastatisk uveal melanom svulst i musen leveren med en høy suksess rate på 80% innen seks måneder. Den xenograft tumor engrafts i leveren som en ensom svulst uten datter knuter (figur 1 og figur 3a). Den kirurgiske orthotopic injeksjon teknikk i leveren bruker microneedles vellykket engrafted kultivert menneskelig lever-met...

Watch this Scientific Journal Video about Generation of a Liver Orthotopic Human Uveal Melanoma Xenograft Platform in Immunodeficient Mice Video at JoVE.com

Generation of a Liver Orthotopic Human Uveal Melanoma Xenograft Platform in Immunodeficient Mice Video (Video) | JoVE

Orthotopic Human lever metastatisk uveal melanom xenograft mus modeller ble opprettet ved hjelp av kirurgiske Orthotopic implantation teknikker med pasient-avledet tumor blings og nål injeksjon teknikker med kultivert menneskelig uveal melanom cellelinjer.

Generering av en Liver Orthotopic Human uveal melanom Xenograft Platform i Immunodeficient mus

In recent decades, subcutaneously implanted patient-derived xenograft tumors or cultured human cell lines have been increasingly recognized as more ...

De fleste pasientavledede tumordyrmodeller implanteres subkutant, og forhindrer evaluering av viktige egenskaper ved svulster og av legemiddeleffekten. Bolig de implanterte svulstene i leveren tilrettelagt denne analysen uten risiko for for mye avvik som oppstår i leveren. Før du begynner prosedyren, spray alle materialer og instrumenter med 70% etylalkohol.Bekreft mangel på respons på tåklemmen i det bedøvede dyret. Plasser musen i liggende stilling på en varmepute, og våt pelsen med ytterligere 70% etylalkohol. Spre pelsen for å visualisere huden under venstre subcostal område.Fjern håret, og påfør jod som skal absorberes i huden. Plasser en steril kirurgisk drapering med et to centimeter hull over musen, og bruk buede ultra fine tang for å løfte bukhuden. Lag en en centimeter tverrgående venstre subcostal snitt inn i telthuden, og sett buede saksspisser inn i snittet for å skille bukhinnen fra huden.Trekk tilbake saksen uten å åpne bladene, og finn leveren under bukhinnen med sin mørke rødlige farge. Bruk den buede saksen til å lage en centimeter tverrgående snitt i bukhinnen. Ved hjelp av den andre hånden, sett kanten av en bomullspinne under venstre leverflibe, og rull vattpinnen nedover for å presse opp leveren.Løft vattpinnen for å overføre leveren på et ikke-vevd, absorberende stoffark. Hold et sterilt nummer 11 skalpellblad parallelt med overflaten, trykk bladet horisontalt inn i leveren for å gjøre en fem millimeter dyp med fem millimeter bredt snitt i parenchyma mens du mykt presser snittet med en bomullspinne. Rull bomullspinnen oppover for å åpne snittstedet.Bruk buede ultra fine tang for å implantere en en millimeter kube av tumorvev i lommen. Trekk inn tangen mens du ruller bomullspinnen i motsatt retning mens du trykker ned, og fjern vattpinnen. Plasser umiddelbart en absorberbar hemostat på snittstedet, og bekreft hemostase.Neste bruk sløv endte tang for å skrelle leveren fra stoffarket, og returnere leveren til bukhulen. Lukk deretter bukhinnen med en 5-0 absorberbar sutur og en dobbel ligatur, og lukk huden med en 5-0 absorberbar sutur og en trippel ligatur. Xenograft svulsten engrafts i leveren som en enslig svulst uten datter knuter.Kirurgisk ortotopisk injeksjon i leveren ved hjelp av mikronegler kan resultere i en vellykket engraftment av en klynge av kultiverte humane levermetastatiske uveale melanomceller i leveren. Ved hjelp av denne leveren ortopiske pasient-avledet tumor modell, effekten av narkotika injisert direkte inn i leveren kan testes med lignende forventede resultater til de som er observert hos kliniske pasienter.

Research

JoVE Journal

Cancer Research

Generation of a Liver Orthotopic Human Uveal Melanoma Xenograft Platform in Immunodeficient Mice

Fluorescent Orthotopic Mouse Model of Pancreatic Cancer

Fluorescent Orthotopic Mouse Model of Pancreatic Cancer Video (Video) | JoVE

Fluorescent ortotopiske musemodell for kreft i bukspyttkjertelen

Pancreatic cancer remains one of the cancers for which survival has not improved substantially in the last few decades. Only 7% of diagnosed patients will ...

Tumor Engraftment in a Xenograft Mouse Model of Human Mantle Cell Lymphoma

Tumor Engraftment in a Xenograft Mouse Model of Human Mantle Cell Lymphoma Video (Video) | JoVE

Svulst Engraftment i en Xenograft musemodell av menneskelig mantelen celle lymfomer

B lymphocytes are key players in immune cell circulation and they mainly home to and reside in lymphoid organs. While normal B cells only proliferate when ...

A 3D Organotypic Melanoma Spheroid Skin Model

A 3D Organotypic Melanoma Spheroid Skin Model Video (Video) | JoVE

3D Organotypic melanom Spheroid huden modell

Malignant transformation of melanocytes, the pigment cells of human skin, causes formation of melanoma, a highly aggressive cancer with increased ...

Orthotopic Injection of Breast Cancer Cells into the Mice Mammary Fat Pad

Orthotopic Injection of Breast Cancer Cells into the Mice Mammary Fat Pad Video (Video) | JoVE

Orthotopic injeksjon av brystkreft celler i mus Mammary fett puten

A proper animal model is crucial for a better understanding of diseases. Animal models established by different methods (subcutaneous injections, ...

Patient-derived Orthotopic Xenograft Models for Human Urothelial Cell Carcinoma and Colorectal Cancer Tumor Growth and Spontaneous Metastasis

Patient-derived Orthotopic Xenograft Models for Human Urothelial Cell Carcinoma and Colorectal Cancer Tumor Growth and Spontaneous Metastasis Video (Video) | JoVE

Pasient-avledet Orthotopic Xenograft modeller for Human Urotelialt Cell kreft og tykktarmskreft tumor vekst og spontan metastasering

Cancer patients have poor prognoses when lymph node (LN) involvement is present in both high-grade urothelial cell carcinoma (HG-UCC) of the bladder and ...

Patient-derived Heterogeneous Xenograft Model of Pancreatic Cancer Using Zebrafish Larvae as Hosts for Comparative Drug Assessment

Patient-derived Heterogeneous Xenograft Model of Pancreatic Cancer Using Zebrafish Larvae as Hosts for Comparative Drug Assessment Video (Video) | JoVE

Pasient-avledet heterogen Xenograft modell av bukspyttkjertelen Cancer bruke sebrafisk larver som verter for sammenlignende Drug Assessment

Patient-derived tumor xenograft (PDX) and cell-derived tumor xenograft (CDX) are important techniques for preclinical assessment, medication guidance and ...

A Melanoma Patient-Derived Xenograft Model

A Melanoma Patient-Derived Xenograft Model Video (Video) | JoVE

En melanom pasient-avledet Xenograft modell

Accumulating evidence suggests that molecular and biological properties differ in melanoma cells grown in traditional two-dimensional tissue culture ...

Establishment of Gastric Cancer Patient-derived Xenograft Models and Primary Cell Lines

Establishment of Gastric Cancer Patient-derived Xenograft Models and Primary Cell Lines Video (Video) | JoVE

Etablering av magekreft pasient-avledet Xenograft modeller og primær celle Lines

The use of preclinical models to advance our understanding of tumor biology and investigate the efficacy of therapeutic agents is key to cancer research. ...

Generation of Orthotopic Pancreatic Tumors and Ex vivo Characterization of Tumor-Infiltrating T Cell Cytotoxicity

Generation of Orthotopic Pancreatic Tumors and Ex vivo Characterization of Tumor-Infiltrating T Cell Cytotoxicity Video (Video) | JoVE

Generering av Orthotopic bukspyttkjertel svulster og ex vivo karakterisering av tumor-infiltrere T Cell cytotoksisitet

In vivo models of pancreatic cancer provide invaluable tools for studying disease dynamics, immune infiltration and new therapeutic strategies. The ...

Spatial Profiling of Protein and RNA Expression in Tissue: An Approach to Fine-Tune Virtual Microdissection

Spatial Profiling of Protein and RNA Expression in Tissue: An Approach to Fine-Tune Virtual Microdissection Video (Video) | JoVE

Romlig profilering av protein- og RNA-uttrykk i vev: En tilnærming til finjustering av virtuell mikrodisseksjon

Multiplexing enables the assessment of several markers on the same tissue while providing spatial context. Spatial Omics technologies allow both protein ...