An agarose spot chemotaxis assay for chemokine receptor antagonists.

Abstract

INTRODUCTION:

Chemokines are important players in directing the migration of cancer cells as part of the metastatic process. The aim of this study is to develop an easy-to-perform, reliable, and inexpensive assay for rapid analysis of anti-chemotactic activity of chemokine antagonists under a number of experimental conditions.

METHODS:

An agarose spot containing the chemokine chemoattractant is applied to a glass petri dish. Live cells in a media, both with and without a chemokine antagonist, are added to the dish and, following cell adhesion, the migration under the agarose spot is observed and analysed by microscopy.

RESULTS:

In the absence of CXCL12 in the agarose, no migration under the agarose spot is detected. In the presence of CXCL12, significant migration under the agarose spot is observed which can be retarded if a neutralising monoclonal antibody or a small molecule antagonist is added to the media.

DISCUSSION:

This experimental configuration is a reliable, inexpensive and easy-to-perform chemotaxis assay, which enables assessment of the activity of CXCR4 antagonists.

• Authors: Victoria Vinader, Yousef Al-Saraireh, Helen L Wiggins, Joshua Z Rappoport, Steve D Shnyder, Laurence H Patterson, Kamyar Afarinkia
• Journal: Journal of pharmacological and toxicological methods (J Pharmacol Toxicol Methods) 2011 Nov-Dec Vol. 64 Issue 3 Pg. 213-6 ISSN: 1873-488X [Electronic] United States
• PMID: 21292017 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2011 Elsevier Inc. All rights reserved.

Chemical References

• Chemokine CXCL12
• Receptors, Chemokine
• Sepharose

Topics

• Adenocarcinoma (diagnosis, immunology)
• Breast Neoplasms (diagnosis, immunology)
• Cell Adhesion (immunology)
• Cell Line, Tumor
• Cell Movement (immunology)
• Chemokine CXCL12 (immunology)
• Chemotaxis (immunology)
• Female
• Humans
• Immunologic Techniques
• Receptors, Chemokine (antagonists & inhibitors, immunology)
• Sepharose (chemistry, immunology)