Technical Advance: Novel ex vivo culture method for human monocytes uses shear flow to prevent total loss of transendothelial diapedesis function

Novel diapedisis method uses shear flow following siRNA knockdown in human monocytes, to improve transendothelial migration. Monocyte recruitment to inflammatory sites and their transendothelial migration into tissues are critical to homeostasis and pathogenesis of chronic inflammatory diseases. How...

Full description

Saved in:
Bibliographic Details
Published in:Journal of leukocyte biology 2014-01, Vol.95 (1), p.191-195
Main Authors: Tsubota, Yoshiaki, Frey, Jeremy M., Raines, Elaine W.
Format: Article
Language:English
Subjects:
macrophages
migration
Technical Advance
transfusion medicine
vascular biology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Novel diapedisis method uses shear flow following siRNA knockdown in human monocytes, to improve transendothelial migration. Monocyte recruitment to inflammatory sites and their transendothelial migration into tissues are critical to homeostasis and pathogenesis of chronic inflammatory diseases. However, even short‐term suspension culture of primary human monocytes leads to phenotypic changes. In this study, we characterize the functional effects of ex vivo monocyte culture on the steps involved in monocyte transendothelial migration. Our data demonstrate that monocyte diapedesis is impaired by as little as 4 h culture, and the locomotion step is subsequently compromised. After 16 h in culture, monocyte diapedesis is irreversibly reduced by ∼90%. However, maintenance of monocytes under conditions mimicking physiological flow (5–7.5 dyn/cm2) is sufficient to reduce diapedesis impairment significantly. Thus, through the application of shear during ex vivo culture of monocytes, our study establishes a novel protocol, allowing functional analyses of monocytes not currently possible under static culture conditions. These data further suggest that monocyte‐based therapeutic applications may be measurably improved by alteration of ex vivo conditions before their use in patients.
ISSN:0741-5400
1938-3673
DOI:10.1189/jlb.0513272