[Dextran sedimentation for study of neutrophil polarization].
OBJECTIVE
To determine the optimal method for separating neutrophils for studying neutrophil polarization.


METHODS
Human neutrophil was separated from healthy human peripheral blood by Percoll density gradient centrifugation and Dextran sedimentation. The cell polarization, purity and activity of the neutrophils were determined, and F-actin polymerization and [Ca2+]i were analyzed.


RESULTS
No significant difference was found in cell polarization, purity and activity of the human neutrophils separated by Dextran sedimentation and Percoll density gradient centrifugation (P>0.05), but F-actin polymerization was inhibited in PMNs separated by Dextran sedimentation, and the peak value of [Ca2+]i was decreased by 25% in PMNs separated by Dextran sedimentation compared to the cells separated by Percoll density gradient centrifugation.


CONCLUSIONS
Both Percoll density gradient centrifugation and Dextran sedimentation can be used for isolating human neutrophils to study cell polarization, but the former method allows better isolation. Dextran sedimentation can be considered when a large number of neutrophils need to be separated.
Luo
Qiong
Q
Department of Occupational Health and Occupational Medicine, School of Public Health, Southern Medical University, Guangzhou 510515, China. luoyy1982@fimmu.com
Cai
Chun-qing
CQ
Xie
Xui-hua
XH
Chu
Xin-wei
XW
Wu
Qing-yi
QY
Meng
Xiao-jing
XJ
Zou
Fei
F
D000199:Actins; D016764:Cell Polarity; D002469:Cell Separation; D002499:Centrifugation, Density Gradient / Q000379:methods*; D006801:Humans; D007958:Leukocyte Count; D009504:Neutrophils / Q000166:cytology*; D011205:Povidone; D012822:Silicon Dioxide
D000199:Actins; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone