[用于研究中性粒细胞极化的葡聚糖沉降法]
[Dextran sedimentation for study of neutrophil polarization].
作者信息
Luo Qiong, Cai Chun-qing, Xie Xui-hua, Chu Xin-wei, Wu Qing-yi, Meng Xiao-jing, Zou Fei
机构信息
Department of Occupational Health and Occupational Medicine, School of Public Health, Southern Medical University, Guangzhou 510515, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2010 Jul;30(7):1514-7.
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.
目的
确定用于研究中性粒细胞极化的中性粒细胞分离的最佳方法。
方法
通过Percoll密度梯度离心法和葡聚糖沉降法从健康人外周血中分离人中性粒细胞。测定中性粒细胞的细胞极化、纯度和活性，并分析F-肌动蛋白聚合和[Ca2+]i。
结果
葡聚糖沉降法和Percoll密度梯度离心法分离的人中性粒细胞在细胞极化、纯度和活性方面无显著差异（P>0.05），但葡聚糖沉降法分离的多形核中性粒细胞中F-肌动蛋白聚合受到抑制，与Percoll密度梯度离心法分离的细胞相比，葡聚糖沉降法分离的多形核中性粒细胞中[Ca2+]i峰值降低了25%。
结论
Percoll密度梯度离心法和葡聚糖沉降法均可用于分离人中性粒细胞以研究细胞极化，但前一种方法分离效果更好。当需要分离大量中性粒细胞时，可考虑葡聚糖沉降法。
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