全血中性粒细胞分离方法对fMLP诱导极性化的影响  

Dextran sedimentation for study of neutrophil polarization

作　　者：罗琼[1] 蔡春青[1] 谢翠华[2] 楚心唯[1] 武清宜[1] 孟晓静[1] 邹飞[1] 

机构地区：[1]南方医科大学公共卫生与热带医学学院职业卫生与职业医学系,广东广州510515 [2]南方医科大学南方医院内分泌代谢科,广东广州510515

出　　处：《南方医科大学学报》2010年第7期1514-1517,共4页Journal of Southern Medical University

基　　金：国家自然科学基金(30971193;30800438;30672353)

摘　　要：目的比较Percoll非连续密度梯度离心法和Dextran沉降法两种常用的中性粒细胞分离方法 ,为中性粒细胞极性化研究提供有效手段。方法取健康成年人外周血,采用Percoll非连续密度梯度离心法和Dextran沉降法分离中性粒细胞,分别检测分离后的中性粒细胞的极性、纯度、存活率,比较二者对极性信号分子F-actin的聚合和胞内Ca2+的影响。结果二者在极性率、纯度、存活率方面并不存在显著性差异,但Dextran沉降法在fMLP诱导细胞极性3min内对F-actin的聚合产生抑制作用,同时可使胞内Ca2+升高的Ca2+峰降低约25%。结论对PMNs极性功能研究时,两种分离方法都可以选择。Percoll非连续密度梯度离心法更有优势,但在需要纯化大量PMNs时,Dextran沉降法也是一种好的选择。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 （P0.05）, 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.

关 键 词：中性粒细胞 极性 Percoll非连续密度梯度离心法 Dextran沉降法 

分 类 号：R392[医药卫生—免疫学]