Name: ex vivo organotypic corneal model of acute epithelial herpes simplex virus type i infection
Uploaded: 2012-11-03T13:00:00
Description: Watch this Scientific Journal Video about Ex Vivo Organotypic Corneal Model of Acute Epithelial Herpes Simplex Virus Type I Infection at JoVE.com

Støtte fra Lions Eye Bank of Delaware Valley var uvurderlig for dette arbeidet. Primære monoklonalt antistoff mot ICP8 var en slags gave fra Dr. David Knipe (Harvard Medical School). Vi vil også takke Dr. Stephen Jennings (Drexel University College of Medicine) og Dr. Peter Laibson (Wills Eye Institute) for personer diskusjoner og kompetanse.

Alle reagenser og utstyr ble kjøpt sterile eller ble sterilisert før prosedyren. Denne artikkelen beskriver fremgangsmåten for å sette opp ex vivo-modellen begynner med en pre-enucleated øyet. I våre eksperimenter, bruker vi fersk hel øyeepler fra unge (8-12 uker) albinokaniner (Pel-Freez Biologicals, Rogers, AR). I tillegg bruker vi menneskelige hornhinner hentet fra lokale øye bank. Hvis du utfører enucleation selv, ta vare å unngå å skade hornhinnen eller Limbus under prosedyren. Rabbit enucleati...

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	<li>Kaye, S., Choudhary, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Herpes+simplex+keratitis.">Herpes simplex keratitis.</a> Prog. Retin. Eye Res. 25, 355-380 (2006).</li><li>Holmberg, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enucleation+of+exotic+pets.">Enucleation of exotic pets.</a> Journal of Exotic Pet Medicine. 16, 88-94 (2007).</li><li>Remeijer, L., Osterhaus, A., Verjans, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+herpes+simplex+virus+keratitis:+the+pathogenesis+revisited.">Human herpes simplex virus keratitis: the pathogenesis revisited.</a> Ocul. Immunol. Inflamm. 12, 255-285 (2004).</li><li>Foreman, D. M., Pancholi, S., Jarvis-Evans, J., McLeod, D., Boulton, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+organ+culture+model+for+assessing+the+effects+of+growth+factors+on+corneal+re-epithelialization.">A simple organ culture model for assessing the effects of growth factors on corneal re-epithelialization.</a> Exp. Eye Res. 62, 555-564 (1996).</li><li>Xu, K. P., Li, X. F., Yu, F. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Corneal+organ+culture+model+for+assessing+epithelial+responses+to+surfactants.">Corneal organ culture model for assessing epithelial responses to surfactants.</a> Toxicol. Sci. 58, 306-314 (2000).</li><li>Klausner, E. A., Peer, D., Chapman, R. L., Multack, R. F., Andurkar, S. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Corneal+gene+therapy.">Corneal gene therapy.</a> J. Control Release. 124, 107-133 (2007).</li><li>Halford, W. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ICP0+antagonizes+Stat+1-dependent+repression+of+herpes+simplex+virus:+implications+for+the+regulation+of+viral+latency.">ICP0 antagonizes Stat 1-dependent repression of herpes simplex virus: implications for the regulation of viral latency.</a> Virol. J. 3, 44 (2006).</li><li>Bhattacharjee, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effective+treatment+of+ocular+HSK+with+a+human+apolipoprotein+E+mimetic+peptide+in+a+mouse+eye+model.">Effective treatment of ocular HSK with a human apolipoprotein E mimetic peptide in a mouse eye model.</a> Invest Ophthalmol Vis. Sci. 49, 4263-4268 (2008).</li><li>Novitskaya, E. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Difficulties+imaging+herpes+simplex+keratitis+with+fluorescein+isothiocynate-labeled+anti-HSV-1+antibodies+in+an+ex+vivo+model.">Difficulties imaging herpes simplex keratitis with fluorescein isothiocynate-labeled anti-HSV-1 antibodies in an ex vivo model.</a> Cornea. 28, 421-425 (2009).</li><li>Sharma, A., Shimeld, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+immunofluorescence+to+diagnose+herpes+simplex+virus+keratitis+in+mice.">In vivo immunofluorescence to diagnose herpes simplex virus keratitis in mice.</a> Br. J. Ophthalmol. 81, 785-788 (1997).</li></ol>

I denne video artikkelen beskriver vi en fremgangsmåte for å studere akutt hornhinneepitel HSV-1 infeksjon i et ex vivo-modell. Denne modellen er en nyttig springbrett mellom in vitro og in vivo-modeller, fordi det gir mulighet for fysiologisk-nøyaktige validering av cellekultur resultater og derved begrenser mengden av dyreforsøk som må utføres. I tillegg gir den ex vivo-modellen en unik og uvurderlig mulighet til å studere epithelial herpes infeksjon i intakte menneskelig...

<table border="1"><tbody><tr><td> Reagenser / rekvisita </td><td> Selskapet </td><td> Katalog # </td></tr><tr><td> Minimum Essential Medium (MEM) </td><td> Mediatech </td><td> 10-010-CV </td></tr><tr><td> Ikke-essensielle aminosyrer (100x løsning) </td><td> Mediatech </td><td> 25-025-CI </td></tr><tr><td> L-Glutamin </td><td> Mediatech </td><td> 25-005-CI </td></tr><tr><td> Penicillin G Sodium Salt </td><td> Sigma </td><td> P3032 </td></tr><tr><td> Streptomycin Sulfate Salt </td><td> Sigma </td><td> S9137 </td></tr><tr><td> Agarose </td><td> Invitrogen </td><td> 15510-027 </td></tr><tr><td> Fosfatbufret saltoppløsning (PBS) </td><td> Utarbeidet i lab </td><td> N / A </td></tr><tr><td> Trypsin EDTA (1x) &lt;/ Td&gt; <td> Mediatech </td><td> 25-053-CI </td></tr><tr><td> DNeasy Blood &amp; Tissue Kit </td><td> QIAGEN </td><td> 69504 </td></tr><tr><td> RNeasy Mini Kit </td><td> QIAGEN </td><td> 74104 </td></tr><tr><td> Laemmli Buffer </td><td> Utarbeidet i lab </td><td> N / A </td></tr><tr><td> Optimal Cutting Temperatur (OCT) Compound </td><td> Tissue-Tek </td><td> 4583 </td></tr><tr><td> Hele kaninøyne (Young) </td><td> Pel-Freez </td><td> 41211-2 </td></tr><tr><td> Menneskelige hornhinner og Whole Eyes </td><td> Lokale Eye Bank </td><td> N / A </td></tr><tr><td> 12 Vel Porcelain Spot Plate </td><td> Avogadros Lab forsyning </td><td> 1877 </td></tr><tr><td> 35 mm Tissue Culture Dish </td><td> Falk </td><td> 353001 </td></tr><tr><td> 6 WellCell Culture Plate </td><td> Greiner Bio-One </td><td> 657160 </td></tr><tr><td> Cryomold Intermediate </td><td> Tissue-Tek </td><td> 4566 </td></tr></tbody></table>

ex vivo organotypic corneal model of acute epithelial herpes simplex virus type i infection

Herpes keratitt er en av de mest alvorlige sykdommer forbundet med herpes simplex virus-type 1 (HSV-1). Herpes keratitt er i dag den ledende årsaken til både hornhinnen-avledet og infeksjon-assosiert blindhet i den industrialiserte verden. Typisk presentasjon av herpes keratitt omfatter infeksjon i hornhinnen epitel og noen ganger dypere hornhinnen stroma og endotel, som fører til slike permanente hornhinnen patologi som arrdannelse, tynning, og opasitet en. Corneal HSV-1-infeksjon er tradisjonelt studert i to typer eksperimentelle modeller. In vitro-modellen, der dyrkede monolag av corneale epitele celler infisert i en petriskål, og tilbyr enkelhet, høy replikerbarhet, raske eksperimenter, og relativt lave kostnader. På den annen side, gir in vivo modell, der dyrene som kaniner eller mus er inokulert direkte i hornhinnen, en meget sofistikert fysiologisksystem, men har høyere kostnader, lengre eksperimenter, nødvendig dyr omsorg, og en større grad av variasjon. I denne videoen artikkelen gir vi en detaljert demonstrasjon av en ny ex vivo modell av hornhinneepitel HSV-1-infeksjon, som kombinerer styrken av både in vitro og in vivo-modeller. Ex vivo Modellen benytter intakt hornhinner organotypically vedlikeholdt i kultur og infisert med HSV-1. Bruken av den ex vivo-modellen muliggjør høyt fysiologisk baserte konklusjoner, men det er ganske billig og krever tid forpliktelse sammenlignbar in vitro modellen.

Watch this Scientific Journal Video about Ex Vivo Organotypic Corneal Model of Acute Epithelial Herpes Simplex Virus Type I Infection at JoVE.com

Ex Vivo Organotypic Corneal Model of Acute Epithelial Herpes Simplex Virus Type I Infection

I denne video artikkelen beskriver vi bruken av en ny<em&gt; Ex vivo</em&gt; Modell av akutte herpes simplex virus type I hornhinneepitel infeksjon.

Ex Vivo Organotypic Corneal Modell av Akutt Epiteliale Herpes simplex virus type I infeksjon

Herpes keratitis is one of the most severe pathologies associated with the herpes simplex virus-type 1 (HSV-1).  Herpes keratitis is currently the leading ...

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Ex Vivo Organotypic Corneal Model of Acute Epithelial Herpes Simplex Virus Type I Infection

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