title: Quantitative Analysis of Dendritic Cell Haptotaxis.
abstract: Chemokines are the main guidance cues directing leukocyte migration. Opposed to early assumptions, chemokines do not necessarily act as soluble cues but are often immobilized within tissues, e.g., dendritic cell migration toward lymphatic vessels is guided by a haptotactic gradient of the chemokine CCL21. Controlled assay systems to quantitatively study haptotaxis in vitro are still missing. In this chapter, we describe an in vitro haptotaxis assay optimized for the unique properties of dendritic cells. The chemokine CCL21 is immobilized in a bioactive state, using laser-assisted protein adsorption by photobleaching. The cells follow this immobilized CCL21 gradient in a haptotaxis chamber, which provides three dimensionally confined migration conditions.
authors.affiliation: Institute of Science and Technology Austria (IST Austria), Klosterneuburg, Austria.
authors.forename: Jan
authors.initials: J
authors.lastname: Schwarz
authors.affiliation: Institute of Science and Technology Austria (IST Austria), Klosterneuburg, Austria. Electronic address: michael.sixt@ist.ac.at.
authors.forename: Michael
authors.initials: M
authors.lastname: Sixt
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology; D003585:Cell Biology / Q000295:instrumentation; D054421:Chemokine CCL21 / Q000737:chemistry / Q000378:metabolism*; D002633:Chemotaxis; D003584:Cytological Techniques / Q000295:instrumentation / Q000379:methods*; D003713:Dendritic Cells / Q000166:cytology*; D006801:Humans; D055767:Immobilized Proteins
chemical_list: C515478:CCL21 protein, human; D054421:Chemokine CCL21; D055767:Immobilized Proteins
keywords: Chemokine; Confinement; Dendritic cell; Gradient; Haptotaxis; Leukocyte; Migration; Surface immobilization