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Collection of in vivo transmigrated neutrophils from human skin.

Review article
Authors Karin Christenson
Lena Björkman
Lisa Davidsson
Anna Karlsson
Per Follin
Claes Dahlgren
Johan Bylund
Published in Methods in molecular biology (Clifton, N.J.)
Volume 1124
Pages 39-52
ISSN 1940-6029
Publication year 2014
Published at Institute of Medicine, Department of Rheumatology and Inflammation Research
Pages 39-52
Language en
Links dx.doi.org/10.1007/978-1-62703-845-...
Subject categories Basic Medicine

Abstract

A wealth of knowledge on the life and death of human neutrophils has been obtained by the in vitro study of isolated cells derived from peripheral blood. However, neutrophils are of main importance, physiologically as well as pathologically, after they have left circulation and transmigrated to extravascular tissues. The journey from blood to tissue is complex and eventful, and tissue neutrophils are in many aspects distinct from the cells left in circulation. Here we describe how to obtain human tissue neutrophils in a controlled experimental setting from aseptic skin lesions created by the application of negative pressure. One protocol enables the direct analysis of the blister content, infiltrating leukocytes as well as exudate fluid, and is a simple method to follow multiple parameters of aseptic inflammation in vivo. Also described is the skin chamber technique, a method based on denuded skin blisters which are subsequently covered by collection chambers filled with autologous serum. Although slightly more artificial as compared to analysis of the blister content directly, the cellular yield of this skin chamber method is sufficient to perform a large number of functional analyses of in vivo transmigrated cells.

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