Name: a preclinical mouse model of osteosarcoma to define the extracellular vesicle-mediated communication between tumor and mesenchymal stem cells
Uploaded: 2018-05-06T12:30:00
Description: Watch this Scientific Journal Video about A Preclinical Mouse Model of Osteosarcoma to Define the Extracellular Vesicle-mediated Communication Between Tumor and Mesenchymal Stem Cells at JoVE.com

S.R. Baglio blev støttet af et fællesskab af Associazione Italiana per la Ricerca sul Cancro (AIRC) medfinansieres af den Europæiske Union. Hertil kommer, har dette projekt modtaget støtte fra EUs Horisont 2020 forskning og innovation program under tilskudsaftalen Marie Sklodowska-Curie ingen 660200 (til S.R. Baglio).

Menneskers fedtvæv for mesenkymale stamceller isoleret hidrører fra department of Plastic Surgery Tergooi Hospital (Hilversum, Holland) efter godkendelse af den institutionelle etiske komité og skriftligt informeret samtykke. Normal god landbrugspraksis-positive adipøst MSCs blev indhentet fra Institut for medicinsk og kirurgisk Sciences for børn og voksne (universitetet i Modena og Reggio Emilia).
Dyreforsøg blev udført i overensstemmelse med den nederlandske lov om dyreforsøg, og pro...

<ol>
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D., Tabi, Z., Clayton, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+exosomes+trigger+fibroblast+to+myofibroblast+differentiation.">Cancer exosomes trigger fibroblast to myofibroblast differentiation.</a> Cancer Res. 70 (23), 9621-9630 (2010).</li><li>Webber, J. 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=Differentiation+of+tumour-promoting+stromal+myofibroblasts+by+cancer+exosomes.">Differentiation of tumour-promoting stromal myofibroblasts by cancer exosomes.</a> Oncogene. 34 (3), 290-302 (2015).</li><li>Zhou, W., 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=Cancer-secreted+miR-105+destroys+vascular+endothelial+barriers+to+promote+metastasis.">Cancer-secreted miR-105 destroys vascular endothelial barriers to promote metastasis.</a> Cancer Cell. 25 (4), 501-515 (2014).</li><li>Whiteside, T., Anastasopoulou, E., Voutsas, I., Papamichail, M., Perez, S., Nunes, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exosomes+and+tumor-mediated+immune+suppression.">Exosomes and tumor-mediated immune suppression.</a> Expert Rev Mol Diagn. 15 (10), 1293-1310 (2016).</li><li>Verweij, F. J., Van Eijndhoven, M. A. J., Middeldorp, J., Pegtel, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+viral+microRNA+exchange+via+exosomes+in+vitro+and+in+vivo.">Analysis of viral microRNA exchange via exosomes in vitro and in vivo.</a> Methods Mol Biol. 1024, 53-68 (2013).</li><li>Baglio, S. R., 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=Human+bone+marrow-+and+adipose-mesenchymal+stem+cells+secrete+exosomes+enriched+in+distinctive+miRNA+and+tRNA+species.">Human bone marrow- and adipose-mesenchymal stem cells secrete exosomes enriched in distinctive miRNA and tRNA species.</a> Stem Cell Res Ther. 6 (1), 127 (2015).</li><li>Naaijkens, B. 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Tumor-udskilles ekstracellulære vesikler (EVs) kan ændre fysiologi af lokal og fjern mesenchymale celler til at generere en tumor-støttende miljø. Her beskriver vi generation af en prækliniske musemodel af osteosarkom, der giver dissektion af de EV-medieret interaktioner mellem tumorceller og mesenkymale stamceller-celler (msc) i vivo. Vi viser, at systemisk injektion af menneskelige tumor EV-uddannet MSCs i mus forsynet med osteosarkom xenografts kraftigt fremmer kræft vækst og metastase dannelse ved at ...

Tumor mikromiljø deltager aktivt i de fleste, hvis ikke alle aspekter af tumordannelse og kræft progression, herunder metastase dannelsen og udviklingen af resistens over for therapeutics1. Dette understreger behovet for prækliniske orthotopic kræft musemodeller, der tillader dissektion af de komplekse tumor-stroma interaktioner forekommer i tumor niche.
Blandt de mange cellulære komponenter af tumor mikromiljø bidrage mesenchymale stamceller (msc) stærkt til kræft progression i flere typer af kræft som brystkræft, prostatakræft, hjernesvulster, myelomatose og osteosarkom2 ,3,4,5,6,7. MSC'er er Multipotente stamceller, der findes i forskellige voksne og føtale væv, herunder knoglemarv, fedtvæv, placenta, navlestrengsblod og andre8,9. Som reaktion på kræft-genereret inflammatoriske signaler, MSCs overflytte mod tumor websteder, indarbejde tumor mikromiljø og i sidste ende udvikle sig til kræft-støtte celler10. Disse kræft-associerede MSCs give væsentlige faktorer (dvs., vækstfaktorer, kemokiner, cytokiner og immunosuppressive mæglere) for tumor progression handler både på tumorcellerne og på de omkringliggende stroma2, 3 , 11 , 12 , 13. selv om fremme af tumor virkningerne af kræft-associerede MSCs er blevet undersøgt i talrige kræftmodeller, de mekanismer, hvorved tumorceller omprogrammere MSC'er til at forme en kræft-fremme niche er dårligt forstået. Her beskriver vi generation af en orthotopic xenograft model, der specifikt giver mulighed for undersøgelse af pro-tumorfremkaldende interaktionen mellem knogle kræftceller og MSCs via ekstracellulære vesikler (EVs).
Evt er afgørende mediatorer af intercellulær kommunikation mellem tumor og stromale celler14. Evt bære funktionelle biomolekyler i cellen i oprindelse, herunder proteiner, lipider og regulerende RNA'er. Når frigivet i det ekstracellulære rum, disse blærer kan tages op af omkringliggende celler eller transporteres til fjerntliggende steder via blod eller lymfe cirkulation, og dybt kan påvirke mål celle adfærd. 15 , 16 , 17 For eksempel udbredelsen af kræft evt af stromale fibroblaster kan resultere i myofibroblast differentiering støtte angiogenese og fremskynde tumor vækst i vivo18,19, internalisering af endotel celler kan stimulere tumor angiogenese og øge vaskulær permeabilitet16,20, og interaktion med immunsystemet celler kan føre til undertrykkelse af antitumor immunrespons21.
Vi viste for nylig, ved hjælp af en bioluminescerende orthotopic xenograft musen model af osteosarkom, at tumorceller frigive store mængder af evt at lynhurtig MSCs at erhverve en pro-tumorfremkaldende og pro-metastatisk fænotype. Denne effekt er på grund af en dramatisk ændring i MSC cytokin udtryk profil (benævnt "MSC uddannelse"), og kan forebygges ved administration af en terapeutisk interleukin-6 receptor (IL-6R) antistof7. Vores arbejde viste, at kræft evt er afgørende modulatorer af MSC adfærd, hvilket giver en begrundelse for mikromiljø-målrettede metoder til at standse osteosarkom progression. Heri, beskriver vi en trinvis protokol for at undersøge EV-medieret tumor-MSC interaktion i vivo. Denne model er beregnet til: 1) specifikt definere kræft EV-inducerede ændringer af MSC adfærd i tumor mikromiljø, 2) vurdere, hvordan denne interaktion bidrager til knogle tumorvækst og metastase dannelse og 3) undersøgelse om at blande sig med EV-medieret krydstale i vivo forhindrer kræft progression.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr >
			<td >Equipment</td>
			<td ></td>
			<td ></td>
			<td ></td>
			<td ></td>
			<td ></td>
			<td ></td>
			<td ></td>
		</tr>
		<tr >
			<td >Ultra Centrifuge</td>
			<td >Beckman</td>
			<td >Optima L-90K</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Rotor SW32Ti</td>
			<td >Beckman</td>
			<td >369650</td>
			<td colspan="4">Referred to in the manuscript as ultra-swinging bucket rotor</td>
			<td></td>
		</tr>
		<tr >
			<td >Transmission electron microscope</td>
			<td >Zeiss</td>
			<td >EM109</td>
			<td>Or similar TEM</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Digital camera</td>
			<td >Nikon</td>
			<td >DMX 1200F</td>
			<td>Or similar camera</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Imaging software TEM&#160;</td>
			<td >Nikon</td>
			<td >ACT-1</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Fluorescence microscope</td>
			<td >Zeiss</td>
			<td >Imager.D2</td>
			<td colspan="2">Or similar Fluorescence microscope</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Imaging software FM</td>
			<td>Zeiss</td>
			<td >ZEN Blue</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Incubator</td>
			<td >Nuaire</td>
			<td >4750E</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Centrifuge</td>
			<td >Hettick</td>
			<td >ROTANTA 460R</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >-80 Freezer</td>
			<td >Thermo electro corporation</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >FACS</td>
			<td >BD</td>
			<td >BD FACScalibur</td>
			<td>Or similar flow cytometer</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Drill</td>
			<td >Ferm</td>
			<td >FCT-300</td>
			<td>With 0.8 mm drill</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >HSS micro twist drills, 0.8 mm</td>
			<td >Proxxon</td>
			<td >28 852</td>
			<td>0.8 mm drill</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >IVIS camera</td>
			<td >Xenogen</td>
			<td >Ivis Lumina</td>
			<td colspan="5">Referred to in the manuscript as bioluminescence camera. Xenogen is now part of Perkin Elmer</td>
		</tr>
		<tr >
			<td >Living image software2.60</td>
			<td >Xenogen / Igor Por</td>
			<td >n.a</td>
			<td colspan="2">Xenogen is now part of Perkin Elmer</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >10 &#181;L Syringe</td>
			<td >Hamilton</td>
			<td >Neuros Model 1701 RN</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Needle: Hamilton RN Needle for Syringe, 26 Gauge, Pointstyle AS, custom length 2 cm</td>
			<td >Hamilton</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Caliper</td>
			<td >Mitutoyo</td>
			<td >G08004463</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Autoclave</td>
			<td >Astell</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Heat Lamp</td>
			<td >Philips</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Culture media</td>
			<td ></td>
			<td ></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Fetal Bovine Serum</td>
			<td >Hyclone</td>
			<td >RYG35912</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Platelet Lysate</td>
			<td >n.a.</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >IMDM medium</td>
			<td >Lonza</td>
			<td >BE12-722F</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >alpha-MEM medium</td>
			<td >Lonza</td>
			<td >BE02-002F</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >DMEM medium</td>
			<td >Lonza</td>
			<td>BE12-614F</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >pen/strep/glutamine</td>
			<td >GIBCO</td>
			<td >10378-016</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >heparin</td>
			<td >LEO</td>
			<td >012866-08</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Trypsin/EDTA (10x)</td>
			<td >GIBCO</td>
			<td >15400-054</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Cells</td>
			<td ></td>
			<td ></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >adipose deriverd MSCs</td>
			<td >n.a.</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >GFP-positive MSCs</td>
			<td >n.a.</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >human fibroblasts</td>
			<td >n.a.</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >143B cells</td>
			<td >ATCC</td>
			<td >CRL-8303</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >FLUC-143B cells</td>
			<td >ATCC</td>
			<td >CRL-8303</td>
			<td>Transduced</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Disposables</td>
			<td ></td>
			<td ></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Culture flasks 175 cm2</td>
			<td >CELLSTAR</td>
			<td >660175</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >50 mL tubes</td>
			<td >Greiner bio-one</td>
			<td >210261</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Freeze tubes</td>
			<td >Thermoscientific</td>
			<td >377224</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Ultra-Clear tubes</td>
			<td >Beckman</td>
			<td >344058</td>
			<td colspan="4">Referred to in the manuscript as ultra-centrifuge tubes</td>
			<td></td>
		</tr>
		<tr >
			<td >0,22 &#181;m filter</td>
			<td >Millex</td>
			<td >SLGV033RS</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >200 mesh Formvar-carbon-coated nickel grids</td>
			<td >EMS (Electron Microscopy Sciences)</td>
			<td ></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >0.5 mL insulin syringes with 29G Needle</td>
			<td >Terumo</td>
			<td >U-100&#160;</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Petri dish</td>
			<td >Sigma - Aldrich</td>
			<td >P7612</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Filter paper&#160;</td>
			<td >Thermo fisher Scientific</td>
			<td >50363215</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Reagents / kits</td>
			<td ></td>
			<td ></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >paraformaldehyde</td>
			<td >Alfa Aeser</td>
			<td >43368.9M</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >PBS</td>
			<td >Braun</td>
			<td >220/12257974/110</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >glutaraldehyde</td>
			<td >EMS (Electron Microscopy Sciences)</td>
			<td >16300</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >uranyl oxalate</td>
			<td >EMS (Electron Microscopy Sciences)</td>
			<td >22510</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >urany acetate</td>
			<td >EMS (Electron Microscopy Sciences)</td>
			<td >22400</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >methyl cellulose</td>
			<td >EMS (Electron Microscopy Sciences)</td>
			<td >1560</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >PKH67</td>
			<td >Sigma</td>
			<td >mini67-1kt</td>
			<td colspan="2">Referred to in the manuscript as GFLD</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >BSA</td>
			<td >Sigma</td>
			<td >A8412</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >CBA - human inflammatory cytokine kit</td>
			<td >BD</td>
			<td >551811</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Formaldehyde 37%</td>
			<td >VWR</td>
			<td >104003100</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Carbon Steel surgical blades</td>
			<td >Swann-Morton</td>
			<td >206</td>
			<td colspan="3">Referred to in the manuscript as surgical knife</td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >anti-human vimentin antibody</td>
			<td >Santa Cruz</td>
			<td >sc-6260</td>
			<td>Clone V9</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Antibody diluent</td>
			<td >DAKO</td>
			<td >S0809</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >HRP-labeled anti mouse IgG antibody</td>
			<td >Life Technologies</td>
			<td >32230</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >DAB-kit</td>
			<td >DAKO</td>
			<td >K500711</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >hematoxyllin</td>
			<td >Sigma</td>
			<td >GHS232</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >EDTA-buffer</td>
			<td >n.a.</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Citrate buffer</td>
			<td >n.a.</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >rabbit polyclonal anti-GFP antibody</td>
			<td >Abcam</td>
			<td >n.a.</td>
			<td>Ab290</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >DAPI&#160;</td>
			<td >Life Technologies</td>
			<td >D1306</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Paracetamol, 120 mg / 5 ml syrup</td>
			<td >Bayer</td>
			<td >n.a.</td>
			<td colspan="2">Sinaspril, paracetamol solution for kids</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Isoflurane 1000 mg/g</td>
			<td >Vumc pharmacy</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >buprenofine hydrochloride, 0.3 mg/ml</td>
			<td >Indivior UK Limited</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >lidocaine-HCL 2%</td>
			<td >Vumc pharmacy</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >70% ethanol</td>
			<td >VWR</td>
			<td >93003.1006</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Tissue glue</td>
			<td >Derma+Flex, formulated medical cyanoacrylate</td>
			<td >Vygon</td>
			<td>LB604060</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Eyedrops: Vidisec Carbogel, 2 mg/ml</td>
			<td >Bausch+Lomb</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >D-luciferin, potassium salt</td>
			<td >Gold Biotechnology</td>
			<td >LUCK-1</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Glass slides</td>
			<td >Thermo scientific</td>
			<td >630-0954</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Stainless steel loops&#160;</td>
			<td >n.a.</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Mice experiments</td>
			<td ></td>
			<td ></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Mice, Hsd:Athymic Nude-Foxn1nu,&#160; female, 6 weeks at arrival, bacterial status conform FELASA</td>
			<td >ENVIGO</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Paper-pulp smart home (cage enrichment)</td>
			<td >Bio Services</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Alpha-dri bedding material</td>
			<td >Shepperd Speciality Papers</td>
			<td >n.a.</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Mouse food: Teklad global 18% protein rodent diet</td>
			<td >ENVIGO</td>
			<td >2918-11416M</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Sutures</td>
			<td >Ethicon</td>
			<td >V926H</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Scissors</td>
			<td >Sigma-Aldrich</td>
			<td >S3146-1EA</td>
			<td>(or similar)</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr >
			<td >Tweezers</td>
			<td >Sigma-Aldrich</td>
			<td >F4142-1EA</td>
			<td>(or similar)</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

a preclinical mouse model of osteosarcoma to define the extracellular vesicle-mediated communication between tumor and mesenchymal stem cells

Inden for tumor mikromiljø bidrage bopæl eller rekrutterede mesenchymale til stamceller (msc) til maligne progression i flere kræft typer. Under indflydelse af specifikke miljømæssige signaler, kan disse voksne stamceller frigive paracrine mæglere fører til accelereret tumorvækst og metastaser. Definere krydstale mellem tumor og MSCs er af primær betydning for at forstå de underliggende kræft progression mekanismer og identificere nye mål for terapeutisk intervention.
Kræftceller producerer store mængder af ekstracellulære vesikler (EVs), hvor dybt kan påvirke funktionsmåden af target-cellerne i svulsten mikromiljø eller på fjerne steder. Tumor evt vedlægge funktionelle biomolekyler, herunder inflammatorisk RNA'er og proteiner, (Kræftpsykologisk), som kan uddanne stromale celler at forbedre funktionen metastatisk kræft celler eller deltage i pre metastatisk niche dannelse. I denne artikel vil beskrive vi udviklingen af en prækliniske kræft musemodel, der giver specifik evaluering af de EV-medieret krydstale mellem tumor og mesenkymale stamceller. Vi beskriver først, oprensning og karakterisering af tumor-udskilles evt og vurderingen af EV internalisering af MSC'er. Vi derefter gøre brug af en multiplex perle-baseret immunassay til at evaluere ændringen af MSC cytokin udtryk profil induceret af kræft evt. Endelig, vi illustrere generation af en bioluminescerende orthotopic xenograft musen model af osteosarkom, der sammenfatter tumor-MSC interaktion, og Vis bidrag af EV-uddannet MSC'er til tumor vækst og metastase dannelse.
Vores model giver mulighed for at definere, hvordan kræft evt forme en tumor-støtte miljø, og at vurdere, om blokaden af de EV-medieret kommunikation mellem tumor og MSCs forhindrer kræft progression.

I denne undersøgelse, vi har undersøgt osteosarkom-udskilles evt evne til at uddanne MSCs hen imod en pro-tumorfremkaldende og pro-metastatisk fænotype. Vi viser, at osteosarkom celler frigive exosome-lignende evt, der er internaliseret af MSC'er. Vi målt ændring af MSC cytokin udtryk profil induceret af kræft EVs, og vurderet effekten af de EV-uddannet MSCs på tumorvækst og metastase dannelse. Den generelle repræsentation af undersøgelsen design er illustreret i <strong class="...

Watch this Scientific Journal Video about A Preclinical Mouse Model of Osteosarcoma to Define the Extracellular Vesicle-mediated Communication Between Tumor and Mesenchymal Stem Cells at JoVE.com

A Preclinical Mouse Model of Osteosarcoma to Define the Extracellular Vesicle-mediated Communication Between Tumor and Mesenchymal Stem Cells

En prækliniske musemodel af osteosarkom at definere den ekstracellulære vesikel-medieret kommunikation mellem Tumor og mesenkymale stamceller

Direkte indsprøjtning af kræft-afledte ekstracellulære vesikler (EVs) fører til omprogrammering af knoglemarv støtte tumor progression; dog er det uklart, hvilke celler der mægle denne effekt. Heri, beskriver vi en trinvis protokol for at undersøge EV-medieret tumor-mesenchymale stamceller (MSC) interaktioner i vivo, afslører en afgørende rolle for EV-uddannet MSCs i metastaser.

Within the tumor microenvironment, resident or recruited mesenchymal stem cells (MSCs) contribute to malignant progression in multiple cancer types. Under ...

Research

JoVE Journal

Cancer Research

Development and Maintenance of a Preclinical Patient Derived Tumor Xenograft Model for the Investigation of Novel Anti-Cancer Therapies

Udvikling og vedligeholdelse af et præklinisk Patient Afledt Tumor xenograftmodel for undersøgelse af Novel Anti-Cancer Therapies

Patient derived tumor xenograft (PDTX) models provide a necessary platform in facilitating anti-cancer drug development prior to human trials. Human tumor ...

A Mimic of the Tumor Microenvironment: A Simple Method for Generating Enriched Cell Populations and Investigating Intercellular Communication

En Mimic af tumor mikromiljø: En simpel metode til at generere berigede Cell Befolkninger og Undersøgelse intercellulær kommunikation

Understanding the early heterotypic interactions between cancer cells and the surrounding non-cancerous stroma is important in elucidating the events ...

The Colon-26 Carcinoma Tumor-bearing Mouse as a Model for the Study of Cancer Cachexia

Den Colon-26 Carcinoma Tumor-bærende mus som en model for Studiet af Cancer Kakeksi

Cancer cachexia is the progressive loss of skeletal muscle mass and adipose tissue, negative nitrogen balance, anorexia, fatigue, inflammation, and ...

Isolation of Circulating Tumor Cells in an Orthotopic Mouse Model of Colorectal Cancer

Isolering af cirkulerende tumorceller i en ortotopisk musemodel af kolorektal cancer

Despite the advantages of easy applicability and cost-effectiveness, subcutaneous mouse models have severe limitations and do not accurately simulate ...

Determining the Optimal Inhibitory Frequency for Cancerous Cells Using Tumor Treating Fields (TTFields)

Determining the Optimal Inhibitory Frequency for Cancerous Cells Using Tumor Treating Fields TTFields

Bestemmelse af den optimale Inhibitorisk Frequency for kræftceller Brug Tumor Behandling Fields (TTFields)

Tumor Treating Fields (TTFields) are an effective treatment modality delivered via the continuous, noninvasive application of low-intensity (1-3 V/cm), ...

Analyzing the Communication Between Monocytes and Primary Breast Cancer Cells in an Extracellular Matrix Extract (ECME)-based Three-dimensional System

Analyzing the Communication Between Monocytes and Primary Breast Cancer Cells in an Extracellular Matrix Extract ECME-based Three-dimensional System

Analysere kommunikationen mellem monocytter og primær brystkræftceller i en ekstracellulære Matrix ekstrakt (ECME)-baseret tre-dimensionelle System

Embedded in the extracellular matrix (ECM), normal and neoplastic epithelial cells intimately communicate with hematopoietic and non-hematopoietic cells, ...

A Method to Define the Effects of Environmental Enrichment on Colon Microbiome Biodiversity in a Mouse Colon Tumor Model

En metode til at definere virkningerne af miljøberigelse på tyktarmen Microbiome biodiversitet i en mus tyktarmen tumoren Model

Several recent studies have illustrated the beneficial effects of living in an enriched environment on improving human disease. In mice, environmental ...

Modeling Osteosarcoma Using Li-Fraumeni Syndrome Patient-derived Induced Pluripotent Stem Cells

Modeling osteosarkom ved hjælp af Li-Fraumeni syndrom Patient-afledte inducerede pluripotente stamceller-celler

Li-Fraumeni syndrome (LFS) is an autosomal dominant hereditary cancer disorder. Patients with LFS are predisposed to a various type of tumors, including ...

Three-Dimensional Bone Extracellular Matrix Model for Osteosarcoma

Tre-dimensionelle knogle ekstracellulære Matrix Model for osteosarkom

Osteosarcoma (OS) is the most common and a highly aggressive primary bone tumor. It is characterized with anatomic and histologic variations along with ...

Capturing Small Molecule Communication Between Tissues and Cells Using Imaging Mass Spectrometry

Indfange små molekyle kommunikation mellem væv og celler ved hjælp af Imaging massespektrometri

Imaging mass spectrometry (IMS) has routinely been applied to three types of samples: tissue sections, spheroids, and microbial colonies. These sample ...