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Studying Leukocyte Recruitment Under Flow Conditions

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Basic Cell Culture Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 946))

Abstract

Leukocyte recruitment from the vasculature occurs under conditions of haemodynamic shear stress. The parallel plate flow chamber apparatus is an in vitro system that is widely used to study leukocyte recruitment under shear conditions. The flow chamber is a versatile tool for examining adhesive interactions, as it can be used to study a variety of adhesive substrates, ranging from monolayers of primary cells to isolated adhesion molecules, and a variety of adhesive particles, ranging from leukocytes in whole blood to antibody-coated latex beads. We describe here methods for studying leukocyte recruitment to cytokine-stimulated, transfected or transduced endothelial cells using both whole blood and isolated leukocyte suspensions. These methods enable multiple parameters to be measured, including the total number of recruited leukocytes, the percentage of leukocytes that are rolling or firmly adherent, and the percentage of leukocytes that have transmigrated. Although these methods are described for interactions between leukocytes and endothelial cells, they are broadly applicable to the study of interactions between many combinations of adhesive substrates and adhesive particles.

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Author information

Authors and Affiliations

  1. Calvin, Phoebe and Joan Snyder Institute for Infection, Immunity and Inflammation, University of Calgary, Calgary, AB, Canada

    Sean A. Parsons, Christophe Jurzinsky, Susan L. Cuvelier & Kamala D. Patel

Authors
  1. Sean A. Parsons
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  2. Christophe Jurzinsky
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  3. Susan L. Cuvelier
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  4. Kamala D. Patel
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Corresponding author

Correspondence to Kamala D. Patel .

Editor information

Editors and Affiliations

  1. , Experimental Therapeutics, BC Cancer Agency, West 10th Ave. 675, Vancouver, V5Z 1L3, British Columbia, Canada

    Cheryl D. Helgason

  2. STEMCELL Technologies, Inc., W. Seventh Avenue 570, Vancouver, V5Z 1B3, British Columbia, Canada

    Cindy L. Miller

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Parsons, S.A., Jurzinsky, C., Cuvelier, S.L., Patel, K.D. (2013). Studying Leukocyte Recruitment Under Flow Conditions. In: Helgason, C., Miller, C. (eds) Basic Cell Culture Protocols. Methods in Molecular Biology, vol 946. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-128-8_18

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  • DOI: https://doi.org/10.1007/978-1-62703-128-8_18

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-127-1

  • Online ISBN: 978-1-62703-128-8

  • eBook Packages: Springer Protocols

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