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

Ce travail a été soutenu par le NCI R01 CA120659 (RS).

1. Un test basé sur Matrigel Tube Formation d&#39;évaluer l&#39;activité Vasculogenic de cellules tumorales <ol><li> Préparation des cellules tumorales et les cellules endothéliales microvasculaires: les cellules tumorales du cerveau telles que les cellules U87, les cellules de mélanome B16F1, le cancer du sein MDA-MB-435, et les cellules du côlon HCT116 ont été cultivées dans du DMEM supplémenté avec 10% de FBS et pénicilline / streptomycine (tous d&#39;Invitrogen ). Cellules endothéliales microvasculai...

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Pour ce test pour réussir, la qualité de Matrigel devrait être testée en premier. Un petit échantillon peut être obtenu à partir de BD Bioscience pour pré-lancer le test en utilisant HMVECs. Produits de lots différents peuvent afficher des qualités dissemblables dans lesquelles certains lots ne fournissent pas une condition optimale pour la formation du tube. Deuxièmement, toutes les bulles doivent être évitées lors d&#39;une aliquote de Matrigel est chargé dans des plaques 96 puits, parce que les bulles ...

<table border="1"><tbody><tr><th> Nom du réactif </th><th> Société </th><th> Numéro de catalogue </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> Le facteur de croissance réduite 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> Microscope Nikon Eclipse TS100 </td><td> Nikon </td><td></td></tr></tbody></table>

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

Au cours des dernières décennies, un essai de formation de tube en utilisant le facteur de croissance réduite Matrigel a été typiquement employées pour démontrer l&#39;activité angiogénique des cellules endothéliales vasculaires in vitro, 1-5. Toutefois, les preuves de plus en plus récemment, a montré que ce test n&#39;est pas limité à tester le comportement des cellules endothéliales vasculaires. Au lieu de cela, il a également été utilisé pour tester la capacité d&#39;un certain nombre de cellules tumorales pour développer un phénotype vasculaire 6-8. Cette capacité est compatible avec leur comportement vasculogenic identifiés chez des animaux xenotransplanted, un processus connu sous le nom vasculogenic mimétisme (VM) 9. Il ya une multitude de preuves démontrant que des cellules tumorales à médiation VM joue un rôle vital dans le développement tumoral, indépendant de l&#39;angiogénèse des cellules endothéliales 6, 10-13. Par exemple, les cellules tumorales ont été trouvés à participer dans le sang perfusé, la formation de canaux vasculaires dans des échantillons de tissus provenant de patients atteints de mélanome et le glioblastome 8, 10, 11. Ici, nous avons décrit ce test réseau tubulaire comme un outil utile dans l&#39;évaluation de l&#39;activité vasculogenic des cellules tumorales. Nous avons constaté que certaines lignées cellulaires tumorales, comme le mélanome B16F1 cellules, les cellules de glioblastome U87, et le cancer du sein MDA-MB-435 cellules sont capables de former des tubules vasculaires, mais certains n&#39;ont pas, comme le cancer du côlon cellules HCT116. Par ailleurs, ce phénotype vasculaire est dépendante du nombre de cellules plaquées sur le Matrigel. Par conséquent, ce test peut servir utilitaire puissant pour dépister le potentiel vasculaire d&#39;une variété de types cellulaires incluant les cellules vasculaires, les cellules tumorales ainsi que d&#39;autres cellules.

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

Un essai de formation de tube est utilisé pour évaluer l&#39;activité vasculaire des cellules tumorales.

Un test basé sur Matrigel Tube Formation d&#39;évaluer l&#39;activité Vasculogenic de cellules tumorales

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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