Name: a matrigel-based tube formation assay to assess the vasculogenic activity of tumor cells
Uploaded: 2011-09-07T13:00:00
Description: Watch this Scientific Journal Video about A Matrigel-Based Tube Formation Assay to Assess the Vasculogenic Activity of Tumor Cells at JoVE.com

Dit werk werd ondersteund door NCI R01 CA120659 (RS).

1. Een Matrigel-Based Tube Formation Assay aan de vasculogene activiteit van tumorcellen beoordelen <ol><li> Voorbereiding van de tumorcellen en microvasculaire endotheelcellen: hersentumor cellen zoals U87-cellen, melanoomcellen B16F1, borstkanker MDA-MB-435, en dikke darm cellen HCT116 werden gekweekt in DMEM aangevuld met 10% FBS en penicilline / streptomycine (alle van Invitrogen ). Endotheel cellijn menselijke microvasculaire endotheelcellen (HMVECs) werden gekweekt in EBM2 medium (Lonza), aangevuld met 1 ug / ml...

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Om deze test te slagen, moet de kwaliteit van Matrigel eerst worden getest. Een klein monster kan worden verkregen bij BD bio-vooraf-run de test met behulp van HMVECs. Andere partij producten kunnen verschillend kwaliteiten waarin sommige percelen niet voorzien in een optimale conditie voor buis vorming. Ten tweede moet elke luchtbellen worden vermeden wanneer een portie van Matrigel wordt geladen in 96-well platen, omdat de bubbels kan tubuli vorming verstoren. Als kleine belletjes zijn te vinden in een goed, kan de Ma...

<table border="1"><tbody><tr><th> Naam van het serum </th><th> Vennootschap </th><th> Catalogus nummer </th></tr><tr><td> DMEM </td><td> Invitrogen </td><td> 11995 </td></tr><tr><td> FBS </td><td> Invitrogen </td><td> 16000-044 </td></tr><tr><td> Groeifactor-gereduceerde Matrigel </td><td> BD Bioscience </td><td> 47743-720 </td></tr><tr><td> EBM2 kit </td><td> Lonza </td><td> CC-3156 </td></tr><tr><td> Nikon ECLIPSE TS100 microscoop </td><td> Nikon </td><td></td></tr></tbody></table>

a matrigel-based tube formation assay to assess the vasculogenic activity of tumor cells

In de afgelopen decennia, een buis formatie test met behulp van groeifactor-gereduceerde Matrigel is meestal gebruikt om de angiogene activiteit van de vasculaire endotheliale cellen aan te tonen in vitro 1-5. Heeft echter recent steeds meer bewijzen zien dat deze test niet beperkt tot de vasculaire endotheliale cellen gedrag voor te testen. In plaats daarvan, maar ook is gebruikt om het vermogen van een aantal tumorcellen aan een vasculaire fenotype 6-8 ontwikkelen, testen. Deze mogelijkheid is in overeenstemming met hun vasculogene het gedrag die in xenotransplanted dieren, een proces dat bekend staat als vasculogene mimicry (VM) 9. Er is een veelheid aan bewijsmateriaal waaruit blijkt dat de tumor cel-gemedieerde VM speelt een vitale rol in de ontwikkeling van tumoren, onafhankelijk van de endotheelcellen angiogenese 6, 10-13. Zo werden de tumorcellen gevonden om deel te nemen in het bloed geperfuseerde, vasculaire kanaal formatie in weefselmonsters van melanoom en glioblastoma patiënten 8, 10, 11. Hier beschreven we deze buisvormige netwerk assay als een nuttig instrument bij de evaluatie van de vasculogene activiteit van de tumorcellen. We ontdekten dat sommige tumor cellijnen zoals melanoom B16F1 cellen, glioblastoma U87 cellen, en borstkanker MDA-MB-435-cellen zijn in staat om vasculaire tubuli vormen, maar sommige niet zoals de dikke darm kanker HCT116 cellen. Bovendien is dit vasculaire fenotype is afhankelijk van aantal cellen uitgeplaat op de Matrigel. Daarom kan deze test dienen als krachtig hulpmiddel om de vasculaire potentieel van een groot aantal celtypen met inbegrip van vasculaire cellen, tumorcellen en andere cellen scherm.

Watch this Scientific Journal Video about A Matrigel-Based Tube Formation Assay to Assess the Vasculogenic Activity of Tumor Cells at JoVE.com

A Matrigel-Based Tube Formation Assay to Assess the Vasculogenic Activity of Tumor Cells

Een buis formatie test wordt gebruikt om de vasculaire activiteit van tumorcellen te evalueren.

Een Matrigel-Based Tube Formation Assay aan de vasculogene activiteit van tumorcellen beoordelen

Over the past several decades, a tube formation assay using growth factor-reduced Matrigel has been typically employed to demonstrate the angiogenic ...

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