Name: a unified methodological framework for vestibular schwannoma research
Uploaded: 2017-06-20T13:00:00
Description: Watch this Scientific Journal Video about A Unified Methodological Framework for Vestibular Schwannoma Research at JoVE.com

Dette arbeidet ble støttet av Nasjonalt institutt for døvhet og andre kommunikasjonsforstyrrelser som gir R01DC015824 (KMS) og T32DC00038 (støtte JES og SD), forsvarsministeriet W81XWH-14-1-0091 (KMS), Bertarelli Foundation (KMS) , Nancy Sayles Day Foundation (KMS), Lauder Tinnitus Research Center (KMS), og Barnes Foundation (KMS).

Skriftlig informert samtykke til innsamling av alle prøver ble oppnådd før operasjonen, og forsøkene ble utført i henhold til etikkloven fra Verdens medisinske forening (Helsinki-erklæringen). Alle deler av studieprotokollen ble godkjent av den institusjonelle gjennomgangskomiteen i Massachusetts Eye and Ear og Massachusetts General Hospital. MERK: Seksjonene 1-7 nedenfor er konstruert for å bli utført i rekkefølge ved mottak av en primær human VS eller GAN-prøve fra operasjonen. ...

<ol>
	<li>Babu, 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=Vestibular+schwannomas+in+the+modern+era:+epidemiology,+treatment+trends,+and+disparities+in+management.">Vestibular schwannomas in the modern era: epidemiology, treatment trends, and disparities in management.</a> J Neurosurg. 119 (1), 121-130 (2013).</li><li>Gal, T. J., Shinn, J., Huang, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+epidemiology+and+management+trends+in+acoustic+neuroma.">Current epidemiology and management trends in acoustic neuroma.</a> Otolaryngol Head Neck Surg. 142 (5), 677-681 (2010).</li><li>Propp, J. M., McCarthy, B. J., Davis, F. 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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Management+of+sporadic+vestibular+schwannoma.">Management of sporadic vestibular schwannoma.</a> Otolaryngol Clin North Am. 48 (3), 407-422 (2015).</li><li>Dilwali, 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=Sporadic+vestibular+schwannomas+associated+with+good+hearing+secrete+higher+levels+of+fibroblast+growth+factor+2+than+those+associated+with+poor+hearing+irrespective+of+tumor+size.">Sporadic vestibular schwannomas associated with good hearing secrete higher levels of fibroblast growth factor 2 than those associated with poor hearing irrespective of tumor size.</a> Otol Neurotol. 34 (4), 748-754 (2013).</li><li>Caye-Thomasen, 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=VEGF+and+VEGF+receptor-1+concentration+in+vestibular+schwannoma+homogenates+correlates+to+tumor+growth+rate.">VEGF and VEGF receptor-1 concentration in vestibular schwannoma homogenates correlates to tumor growth rate.</a> Otol Neurotol. 26 (1), 98-101 (2005).</li><li>Koutsimpelas, D., 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=The+VEGF/VEGF-R+axis+in+sporadic+vestibular+schwannomas+correlates+with+irradiation+and+disease+recurrence.">The VEGF/VEGF-R axis in sporadic vestibular schwannomas correlates with irradiation and disease recurrence.</a> ORL J Otorhinolaryngol Relat Spec. 74 (6), 330-338 (2012).</li><li>Dilwali, S., Roberts, D., Stankovic, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interplay+between+VEGF-A+and+cMET+signaling+in+human+vestibular+schwannomas+and+schwann+cells.">Interplay between VEGF-A and cMET signaling in human vestibular schwannomas and schwann cells.</a> Cancer Biol Ther. 16 (1), 170-175 (2015).</li><li>Dilwali, S., Kao, S. Y., Fujita, T., Landegger, L. D., Stankovic, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonsteroidal+anti-inflammatory+medications+are+cytostatic+against+human+vestibular+schwannomas.">Nonsteroidal anti-inflammatory medications are cytostatic against human vestibular schwannomas.</a> Transl Res. 166 (1), 1-11 (2015).</li><li>Dilwali, 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=Preclinical+validation+of+anti-nuclear+factor-kappa+B+therapy+to+inhibit+human+vestibular+schwannoma+growth.">Preclinical validation of anti-nuclear factor-kappa B therapy to inhibit human vestibular schwannoma growth.</a> Mol Oncol. 9 (7), 1359-1370 (2015).</li><li>Dilwali, S., Landegger, L. D., Soares, V. Y., Deschler, D. G., Stankovic, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Secreted+Factors+from+Human+Vestibular+Schwannomas+Can+Cause+Cochlear+Damage.">Secreted Factors from Human Vestibular Schwannomas Can Cause Cochlear Damage.</a> Sci Rep. 5, 18599 (2015).</li><li>Blakeley, J. <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+drug+treatments+for+neurofibromatosis+type+2-associated+vestibular+schwannoma.">Development of drug treatments for neurofibromatosis type 2-associated vestibular schwannoma.</a> Curr Opin Otolaryngol Head Neck Surg. 20 (5), 372-379 (2012).</li><li>Schroeder, R. D., Angelo, L. S., Kurzrock, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NF2/merlin+in+hereditary+neurofibromatosis+2+versus+cancer:+biologic+mechanisms+and+clinical+associations.">NF2/merlin in hereditary neurofibromatosis 2 versus cancer: biologic mechanisms and clinical associations.</a> Oncotarget. 5 (1), 67-77 (2014).</li><li>Schularick, N. M., Clark, J. J., Hansen, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Primary+culture+of+human+vestibular+schwannomas.">Primary culture of human vestibular schwannomas.</a> J Vis Exp. (89), (2014).</li><li>Dilwali, 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=Primary+culture+of+human+Schwann+and+schwannoma+cells:+improved+and+simplified+protocol.">Primary culture of human Schwann and schwannoma cells: improved and simplified protocol.</a> Hear Res. , 25-33 (2014).</li><li>Brown, C. M., Ahmad, Z. K., Ryan, A. F., Doherty, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estrogen+receptor+expression+in+sporadic+vestibular+schwannomas.">Estrogen receptor expression in sporadic vestibular schwannomas.</a> Otol Neurotol. 32 (1), 158-162 (2011).</li><li>Cioffi, J. 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=MicroRNA-21+overexpression+contributes+to+vestibular+schwannoma+cell+proliferation+and+survival.">MicroRNA-21 overexpression contributes to vestibular schwannoma cell proliferation and survival.</a> Otol Neurotol. 31 (9), 1455-1462 (2010).</li><li>Doherty, J. K., Ongkeko, W., Crawley, B., Andalibi, A., Ryan, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ErbB+and+Nrg:+potential+molecular+targets+for+vestibular+schwannoma+pharmacotherapy.">ErbB and Nrg: potential molecular targets for vestibular schwannoma pharmacotherapy.</a> Otol Neurotol. 29 (1), 50-57 (2008).</li><li>Aguiar, P. H., Tatagiba, M., Samii, M., Dankoweit-Timpe, E., Ostertag, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+comparison+between+the+growth+fraction+of+bilateral+vestibular+schwannomas+in+neurofibromatosis+2+(NF2)+and+unilateral+vestibular+schwannomas+using+the+monoclonal+antibody+MIB+1.">The comparison between the growth fraction of bilateral vestibular schwannomas in neurofibromatosis 2 (NF2) and unilateral vestibular schwannomas using the monoclonal antibody MIB 1.</a> Acta Neurochir (Wien). 134 (1-2), 40-45 (1995).</li><li>Cattoretti, G., 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=Monoclonal+antibodies+against+recombinant+parts+of+the+Ki-67+antigen+(MIB+1+and+MIB+3)+detect+proliferating+cells+in+microwave-processed+formalin-fixed+paraffin+sections.">Monoclonal antibodies against recombinant parts of the Ki-67 antigen (MIB 1 and MIB 3) detect proliferating cells in microwave-processed formalin-fixed paraffin sections.</a> J Pathol. 168 (4), 357-363 (1992).</li><li>Hung, G., 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=Immunohistochemistry+study+of+human+vestibular+nerve+schwannoma+differentiation.">Immunohistochemistry study of human vestibular nerve schwannoma differentiation.</a> Glia. 38 (4), 363-370 (2002).</li><li>Archibald, D. J., 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=B7-H1+expression+in+vestibular+schwannomas.">B7-H1 expression in vestibular schwannomas.</a> Otol Neurotol. 31 (6), 991-997 (2010).</li><li>Landegger, L. D., 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+synthetic+AAV+vector+enables+safe+and+efficient+gene+transfer+to+the+mammalian+inner+ear.">A synthetic AAV vector enables safe and efficient gene transfer to the mammalian inner ear.</a> Nat Biotechnol. 35 (3), 280-284 (2017).</li><li>Zinn, 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=In+Silico+Reconstruction+of+the+Viral+Evolutionary+Lineage+Yields+a+Potent+Gene+Therapy+Vector.">In Silico Reconstruction of the Viral Evolutionary Lineage Yields a Potent Gene Therapy Vector.</a> Cell Rep. 12 (6), 1056-1068 (2015).</li><li>Kim, B. G., 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=Sulforaphane,+a+natural+component+of+broccoli,+inhibits+vestibular+schwannoma+growth+in+vitro+and+in+vivo.">Sulforaphane, a natural component of broccoli, inhibits vestibular schwannoma growth in vitro and in vivo.</a> Sci Rep. 6, 36215 (2016).</li><li>Soares, V. 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=Extracellular+vesicles+derived+from+human+vestibular+schwannomas+associated+with+poor+hearing+damage+cochlear+cells.">Extracellular vesicles derived from human vestibular schwannomas associated with poor hearing damage cochlear cells.</a> Neuro Oncol. 18 (11), 1498-1507 (2016).</li><li>Lysaght, A. 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=Proteome+of+human+perilymph.">Proteome of human perilymph.</a> J Proteome Res. 10 (9), 3845-3851 (2011).</li><li>Caye-Thomasen, P., Borup, R., Stangerup, S. E., Thomsen, J., Nielsen, F. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deregulated+genes+in+sporadic+vestibular+schwannomas.">Deregulated genes in sporadic vestibular schwannomas.</a> Otol Neurotol. 31 (2), 256-266 (2010).</li><li>Schulz, 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=The+importance+of+nerve+microenvironment+for+schwannoma+development.">The importance of nerve microenvironment for schwannoma development.</a> Acta Neuropathol. 132 (2), 289-307 (2016).</li><li>Torres-Martin, 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=Global+profiling+in+vestibular+schwannomas+shows+critical+deregulation+of+microRNAs+and+upregulation+in+those+included+in+chromosomal+region+14q32.">Global profiling in vestibular schwannomas shows critical deregulation of microRNAs and upregulation in those included in chromosomal region 14q32.</a> PLoS One. 8 (6), e65868 (2013).</li></ol>

Dette manuskriptet beskriver et enhetlig metodologisk rammeverk for VS-forskning, som beskriver samtidig behandling av menneskelige VS- og GAN-prøver for nedstrøms forskningsapplikasjoner. Som VS-forskning går inn i en alder av presisjonsmedisin, vil det bli mulig å oppdage molekylær, cellulær, genetisk og proteomisk innsikt som er spesifikke for individuelle pasienter, og forberede den samme prøven i skjemaer som kan svare på en rekke forskningsspørsmål. Renheten av humane Schwann...

Vestibulære schwannomer (VS) er de vanligste neoplasmaene i cerebellopontinvinkelen, diagnostisert hos 1 av hver 100.000 individer. Selv om ikke-metastatisk, påvirker disse svulstene alvorlig pasientens livskvalitet 1 , 2 , 3 , 4 , 5 , 6 . Berørte personer lever vanligvis med hørselstap, tinnitus og en følelse av lydstyrke. Symptomene blir stadig svekkende som tumoren vokser, forårsaker balanseproblemer, ansiktslammelse og svekkelse av andre kraniale nervefunksjoner. Livstruende komplikasjoner på grunn av hjernestamkompresjon kan også følge 7 . Administrasjonsmuligheter for VS er i hovedsak begrenset til vaktfull venter på statiske svulster og stereotaktisk strålebehandling eller kirurgisk reseksjon for voksende svulster <sOpp klasse = &quot;xref&quot;&gt; 8. Den kirurgiske fjerning av disse svulstene i forskningsrelaterte sykehus gir mulighet til å erverve og analysere friskt tumorvæv samlet under pasientoperasjoner. En spesifikk kirurgisk tilnærming til VS, den translabyrintiske reseksjonen, kan til og med gi tilgang til verdifull menneskelig sensorisk epitel fra indre øre og perilimf. Fordi VS oppstår fra en perifer sensorisk nerve ( dvs. vestibulærnerven), er det viktig å sammenligne VS-assosierte observasjoner med de som er avledet fra en passende kontrollnerve, slik som den menneskelige store auricularnerven (GAN). Sunn GAN blir regelmessig ofret under parotidektomi eller nakke-disseksjoner og kan brukes som robuste modeller for sunn Schwann-cellefysiologi 9 . Fordi det ikke finnes FDA-godkjente stoffer for behandling eller forebygging av sporadisk VS, er det avgjørende at forskere belyser den underliggende molekylære mechaNismer av sykdommen for å identifisere terapeutiske mål. Proteiner som har vist seg å spille en rolle i VS-patogenesen inkluderer merlin, vaskulær endotelvekstfaktor (VEGF), cyklooksygenase 2 (COX-2), nukleær faktor kappa B (NF-KB), tumornekrosefaktor alfa (TNF-alfa) , Epidermal vekstfaktorreseptor (EGFR) og tilhørende signalmolekyler 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 . Nylige fremskritt har utvidet og forbedret protokoller for innsamling, behandling, kultur og nedstrøms undersøkelse av primære humane vestibulære schwannomer og friske nervevev 18 , 19 . Imidlertid er de fleste eksisterende protokoller designet for å imøtekomme preparatetAtion av slike vev for en enkelt nedstrøms forskningsapplikasjon ( dvs. cellekultur alene). Denne artikkelen presenterer et enhetlig metodologisk rammeverk for samtidig behandling av en enkelt primær menneskelig VS- eller GAN-prøve for flere nedstrømsapplikasjoner: celletypespesifikk kultur, proteinutvinning, RNA-bevaring, svulstsekresjonsoppsamling og vevfiksering. Dette arbeidet beskriver også kirurgisk innsamling og behandling av human cerebrospinalvæske (CSF) og perilimf under translabyrint VS-reseksjon, da disse nært besluttede vevene kan tjene som viktige kilder til biomarkører for VS. Endelig presenterer denne protokollen trinn for viral transduksjon av primære humane VS-celler i kultur som en ny anvendelse av dette vevet for bruk i genterapi.

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		<tr height="42">
			<td height="42" style="height:42px;width:419px;">BioCoat Poly-D-Lysine/Laminin 12mm #1 German Glass Coverslip</td>
			<td style="width:159px;">Corning</td>
			<td style="width:159px;">354087</td>
			<td style="width:683px;">Or prepare coverslips with Corning Laminin (CB-40232) and Cultrex Poly-L-Lysine (3438-100-01)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:419px;">CELLSTAR 15 ml Centrifuge Tubes, Conical bottom, Graduation, Sterile</td>
			<td style="width:159px;">Greiner Bio-One</td>
			<td style="width:159px;">188161</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:419px;">CELLSTAR 50 ml Centrifuge Tubes, Conical bottom, Graduation, Sterile</td>
			<td style="width:159px;">Greiner Bio-One</td>
			<td style="width:159px;">227261</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">CELLSTAR Cell Culture Dish, 60 mm</td>
			<td style="width:159px;">Greiner Bio-One</td>
			<td style="width:159px;">628160</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Collagenase from Clostridium histolyticum, Sterile-filtered</td>
			<td style="width:159px;">Sigma-Aldrich</td>
			<td style="width:159px;">C1639</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Costar 24 Well Clear TC-Treated Multiple Well Plates, Sterile</td>
			<td style="width:159px;">Corning</td>
			<td style="width:159px;">3526</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">DAPI (4&#39;,6-Diamidino-2-Phenylindole, Dihydrochloride)</td>
			<td style="width:159px;">Thermo Fisher Scientific</td>
			<td style="width:159px;">D1306</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:419px;">DMEM, high glucose, pyruvate, no glutamine, 500 ml</td>
			<td style="width:159px;">Thermo Fisher Scientific</td>
			<td style="width:159px;">10313-039</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:419px;">DMEM/F-12, 500 ml</td>
			<td style="width:159px;">Thermo Fisher Scientific</td>
			<td style="width:159px;">11320-033</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Dumont #3 Forceps, Dumoxel</td>
			<td style="width:159px;">Fine Science Tools</td>
			<td style="width:159px;">11231-30</td>
			<td>Autoclave prior to use</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Dumont #5 Forceps, Standard tip, Inox</td>
			<td style="width:159px;">Fine Science Tools</td>
			<td style="width:159px;">11251-20</td>
			<td>Autoclave prior to use</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:419px;">Fetal Bovine Serum, qualified, USDA-approved regions, 500 ml</td>
			<td style="width:159px;">Thermo Fisher Scientific</td>
			<td style="width:159px;">10437-028&#160;</td>
			<td>Aliquot in 50 ml tubes and store in -20&#176;C freezer</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:419px;">Hyaluronidase from Bovine Testes, Type I-S, Lyophilized Powder</td>
			<td style="width:159px;">Sigma-Aldrich</td>
			<td style="width:159px;">H3506</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:419px;">Millex-GP Syringe Filter Unit, 0.22 &#181;m, polyethersulfone, 33 mm, sterile</td>
			<td style="width:159px;">EMD Millipore</td>
			<td style="width:159px;">SLGP033RS</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Paraformaldehyde, Reagent Grade, Crystalline</td>
			<td style="width:159px;">Sigma-Aldrich</td>
			<td style="width:159px;">P6148</td>
			<td>Prior to use: Establish Standard Operating Procedures based on protocols available online</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:419px;">PBS, pH 7.4, 500 ml</td>
			<td style="width:159px;">Thermo Fisher Scientific</td>
			<td style="width:159px;">10010-023&#160;</td>
			<td>Autoclave prior to use</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:419px;">Penicillin-Streptomycin (10,000 U/ml), 100 ml</td>
			<td style="width:159px;">Thermo Fisher Scientific</td>
			<td style="width:159px;">15140-122</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">PhosSTOP Phosphatase Inhibitor Tablets</td>
			<td style="width:159px;">Roche</td>
			<td style="width:159px;">04906845001</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:419px;">Pierce Protease Inhibitor Tablets</td>
			<td style="width:159px;">Thermo Fisher Scientific</td>
			<td style="width:159px;">88666</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Pipettes and pipette tips, 5/10/25 ml</td>
			<td style="width:159px;">Variable</td>
			<td style="width:159px;">Variable</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Plastic Homogenization Pestle for 1.5/2.0ml Microtubes</td>
			<td style="width:159px;">E&#38;K Scientific</td>
			<td style="width:159px;">EK-10539</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">PrecisionGlide Needles, 27 G x 1 1/2 in&#160;</td>
			<td style="width:159px;">BD</td>
			<td style="width:159px;">301629</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">RIPA Buffer</td>
			<td style="width:159px;">Boston BioProducts</td>
			<td style="width:159px;">BP-115</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:419px;">RNAlater (RNA stabilization solution)</td>
			<td style="width:159px;">Thermo Fisher Scientific</td>
			<td style="width:159px;">AM7021</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Safe-Lock Microcentrifuge Tubes, Polypropylene, 0.5 ml</td>
			<td style="width:159px;">Eppendorf</td>
			<td style="width:159px;">022363719</td>
			<td>Autoclave prior to use</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Safe-Lock Microcentrifuge Tubes, Polypropylene, 1.5 ml</td>
			<td style="width:159px;">Eppendorf</td>
			<td style="width:159px;">022363204</td>
			<td>Autoclave prior to use</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Saline - 0.9% Sodium Chloride Injection, bacteriostatic, 20 ml</td>
			<td style="width:159px;">Hospira</td>
			<td style="width:159px;">0409-1966-05</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Scalpel Blades - #15</td>
			<td style="width:159px;">Fine Science Tools</td>
			<td style="width:159px;">10015-00</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Schuknecht Suction Tube 24 gauge</td>
			<td style="width:159px;">Bausch + Lomb</td>
			<td style="width:159px;">N1698 42</td>
			<td>Useful for the surgical approach (in addition to common otologic surgical instruments) and e.g. a blue surgical marker</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Specimen Container, OR sterile, 4OZ&#160;</td>
			<td style="width:159px;">Medline</td>
			<td style="width:159px;">DYND30331H</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Stemi 2000-C Stereo Microscope</td>
			<td style="width:159px;">Zeiss</td>
			<td style="width:159px;">000000-1106-133</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Syringe/Needle Combination, Luer-Lok Tip, 5 ml, 22 G x 1 in.</td>
			<td style="width:159px;">BD</td>
			<td style="width:159px;">309630</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Tuberculin Syringe Only, Slip tip, 1 ml</td>
			<td style="width:159px;">BD</td>
			<td style="width:159px;">309659</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Tuberculin Syringe Only, Slip tip, 3 ml</td>
			<td style="width:159px;">BD</td>
			<td style="width:159px;">309656</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Ultrasonic homogenizer, 4710 Series, CV18 probe</td>
			<td style="width:159px;">Cole-Parmer</td>
			<td style="width:159px;">CP25013</td>
			<td></td>
		</tr>
	</tbody>
</table>

a unified methodological framework for vestibular schwannoma research

Vestibulære schwannomer er de vanligste neoplasmaene i cerebellopontinvinkelen, og utgjør 6-8% av alle intrakranielle vekst. Selv om disse svulstene forårsaker sensorineuralt hørselstap i opptil 95% av de berørte individer, forblir de molekylære mekanismene som ligger til grunn for dette hørselstapet unnvikende. Denne artikkelen skisserer trinnene etablert i vårt laboratorium for å lette oppsamlingen og behandlingen av forskjellige primære humane vevsprøver for nedstrøms forskningsapplikasjoner integrert i studiet av vestibulære schwannomer. Spesielt beskriver dette arbeidet et enhetlig metodologisk rammeverk for innsamling, behandling og kultur av Schwann- og schwannomaceller fra kirurgiske prøver. Dette er integrert med parallelle behandlingsstrinn som nå anses som essensielt for dagens forskning: innsamling av svulst- og nervesekresjoner, bevaring av RNA og ekstraksjon av protein fra oppsamlede vev, fiksering av vev for fremstilling av seksjoner, enD eksponering av primære humane celler til adeno-assosierte virus for anvendelse på genterapi. I tillegg fremhever dette arbeidet den translabyrintiske kirurgiske tilnærmingen for å samle denne svulsten som en unik mulighet til å skaffe menneskelig sensorisk epitel fra indre øre og perilimf. Tips for å forbedre eksperimentell kvalitet er gitt og vanlige fallgruver uthevet.

Primære humane VS-celler i kultur, som etablert i avsnitt 5, kan behandles som informative modeller for sykdomsassosierte prosesser i mange nedstrøms forskningsapplikasjoner ( Figur 1 ). Sunn Schwann-celler dyrket i seksjon 6 gir et direkte og lærerikt sammenligningspunkt. Som beskrevet nedenfor, er omfattende data fra VS og GANer behandlet i henhold til dette enhetlige metodiske ramme tilgjengelig i flere artikler tidligere publisert 12...

Watch this Scientific Journal Video about A Unified Methodological Framework for Vestibular Schwannoma Research at JoVE.com

A Unified Methodological Framework for Vestibular Schwannoma Research

Målet med denne protokollen er å skissere samlingen og behandlingen av menneskelige kirurgiske prøver for flere nedstrømsapplikasjoner i vestibulær schwannom og Schwann-cellforskning.

Et enhetlig metodologisk rammeverk for Vestibular Schwannoma Research

Vestibular schwannomas are the most common neoplasms of the cerebellopontine angle, making up 6-8% percent of all intracranial growths. Though these ...

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