Screening Assays to Characterize Novel Endothelial Regulators Involved in the Inflammatory Response

Mar&#237;a &#193;ngeles Higueras; Lidia Jim&#233;nez-Garc&#237;a; Sandra Herranz; Sonsoles Hortelano; Alfonso Luque

doi:10.3791/55824

Screening Assays to Characterize Novel Endothelial Regulators Involved in the Inflammatory Response

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

Screening Assays to Characterize Novel Endothelial Regulators Involved in the Inflammatory Response

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12:50 min

September 15, 2017

DOI: 10.3791/55824-v

María Ángeles Higueras¹, Lidia Jiménez-García¹, Sandra Herranz¹, Sonsoles Hortelano*¹, Alfonso Luque*¹

¹Pharmacological Therapies Unit, Research Institute for Rare Diseases,Institute of Health Carlos III

Vascular endothelium tightly controls leukocyte recruitment. Inadequate leukocyte extravasation contributes to human inflammatory diseases. Therefore, searching for novel regulatory elements of endothelial activation is necessary to design improved therapies for inflammatory disorders. Here, we describe a comprehensive methodology to characterize novel endothelial regulators that can modify leukocyte trafficking during inflammation.

The overall goal of this screening assay is to characterize novel endothelial regulators involved in the inflammatory response of interest for designing new therapeutic strategies against inflammatory disorders.This method can help answer key questions in the pathology field such as defining novel endothelial-targeting drugs able to attenuate inflammatory-related diseases.The main advantage of this endothelial protocol is that describes assay technology that establishes the basis for exploring novel mechanisms involved in the regulation of the inflammatory response.Demonstrating this methodology will be Maria Angeles Higueras, a post-doc from our lab and Sandra Herranz, a technician from our lab.To begin, culture MLEC-04 cells in complete DMEM F12 medium supplemented with 10%FBS and Pen-Strep.Incubate the cells in a biological incubator at 37

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