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

Diese Arbeit wurde vom NCI R01 CA120659 (RS) unterstützt.

1. Ein Matrigel-Based Rohr Formation Assay zur vaskulären Aktivität von Tumorzellen Beurteilen <ol><li> Vorbereitung der Tumorzellen und mikrovaskulären Endothelzellen: Hirntumor-Zellen wie U87-Zellen, Melanomzellen B16F1, Brustkrebs MDA-MB-435, und die Zellen des Darms HCT116 wurden in DMEM ergänzt mit 10% FBS und Penicillin / Streptomycin (alle von Invitrogen ). Endothelial Zelllinie humanen mikrovaskulären Endothelzellen (HMVECs) wurden in EBM2 Medium (Lonza) mit 1 pg / ml Hydrocortison und 1 ng / ml epidermal...

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Um für diesen Test erfolgreich zu sein, sollte die Qualität der Matrigel zuerst getestet werden. Eine kleine Probe von BD Bioscience erhalten werden, um Vorlauf der Test unter Verwendung von HMVECs. Verschiedene Produkte können Batch-Display ungleichen Qualitäten, in denen einige viel nicht bieten eine optimale Voraussetzung für tube formation. Zweitens sollten alle Blasen vermieden werden, wenn ein Aliquot von Matrigel in 96-Well-Platten geladen werden, da Blasen Tubulus Bildung stören können. Wenn kleine Bläsc...

<table border="1"><tbody><tr><th> Name des Reagenzes </th><th> Firma </th><th> Katalog-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> Growth factor-reduziert 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 Mikroskop </td><td> Nikon </td><td></td></tr></tbody></table>

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

Im Laufe der letzten Jahrzehnte, mit einem Rohr-Assay growth factor-reduzierten Matrigel wurde in der Regel eingesetzt, um die angiogene Aktivität des vaskulären endothelialen Zellen in vitro 1-5 demonstrieren. Allerdings hat vor kurzem immer mehr Belege dafür ergeben, dass dieser Test nicht beschränkt auf vaskuläre Verhalten für Endothelzellen zu testen. Stattdessen hat sie auch benutzt worden, um die Fähigkeit einer Reihe von Tumorzellen zu einer vaskulären Phänotyp 6-8 Entwicklung zu testen. Diese Fähigkeit wurde in Übereinstimmung mit ihren vaskulogene Verhalten in xenotransplantierten Tiere, ein Verfahren, wie vaskulogene Mimikry (VM) 9 bekannt sind, identifiziert. Es gibt eine Vielzahl von Nachweisen dafür, dass Tumor-Zell-vermittelten VM spielt eine wichtige Rolle bei der Tumorentwicklung, unabhängig von Endothelzellen Angiogenese 6, 10-13. So wurden zum Beispiel Tumorzellen im Blut durchströmt, vaskuläre Kanalbildung in Gewebeproben aus Melanomen und Glioblastom-Patienten 8, 10, 11 zu beteiligen. Hier haben wir beschrieben, diese röhrenförmigen Netz-Assay als ein nützliches Instrument bei der Auswertung von vaskulären Aktivität von Tumorzellen. Wir fanden, dass einige Tumorzelllinien wie Melanom B16F1 Zellen, Glioblastom U87-Zellen, und Brustkrebs MDA-MB-435 Zellen in der Lage, vaskuläre Tubuli bilden; einige aber auch nicht wie Darmkrebs HCT116-Zellen. Darüber hinaus ist dieses vaskulären Phänotyp abhängig Zellzahlen auf dem Matrigel beschichtet. Daher kann dieser Test als leistungsstarkes Dienstprogramm für die vaskuläre Potenzial einer Vielzahl von Zelltypen einschließlich vaskulärer Zellen, Tumorzellen sowie anderen Zellen Bildschirm dienen.

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

Ein Rohr-Assay wird verwendet, um vaskuläre Aktivität von Tumorzellen zu bewerten.

Ein Matrigel-Based Rohr Formation Assay zur vaskulären Aktivität von Tumorzellen Beurteilen

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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