ALTERED EXPRESSION OF CD11B/CD18 AND CD62L ON HUMAN MONOCYTES AFTER CELL PREPARATION PROCEDURES
ALTERED EXPRESSION OF CD11B/CD18 AND CD62L ON HUMAN MONOCYTES AFTER CELL PREPARATION PROCEDURES
复制标题
DOI：
10.1016/0022-1759(94)00303-e
复制
发表时间：
1995-03-13
影响因子：
2.2
通讯作者：
HED, J
中科院分区：
文献类型：
--
作者：
LUNDAHL, J;HALLDEN, G;HED, J
我们研究了不同的细胞制备工艺对人单核细胞表面CD11b/CD18和CD62L表达的影响。EDTA抗凝血和肝素抗凝血均作为白细胞的来源。在氯化铵介导的溶解后获得的混合白细胞悬浮液和在4℃和20℃下进行密度梯度离心（Ficoll-Paque）制备的单核细胞悬浮液中，流式细胞术分析了单核细胞。来自肝素化血液的单核细胞比来自EDTA血液的单核细胞具有更高的CD11b/CD18表达和更明显的个体间变异。通过改变CD11b/CD18和CD62L的表达，Ficoll-Paque分离的单核细胞比通过氯化铵介导的裂解制备的单核细胞具有更高的体外活化程度。当分离过程在20℃下进行时，这一点更为明显。我们的研究结果表明，通过氯化铵介导的EDTA血液裂解和保持在4℃的细胞处理温度制备的单核细胞通过受体改变受到最小的体外调节。对不同细胞制备程序的体外调节的认识是重要的，特别是在比较不同来源的白细胞上功能受体的表达时。
We have investigated the effect of different cell preparation procedures on the surface expression of CD11b/CD18 and CD62L on human monocytes. Both EDTA and heparin anticoagulated blood were used as sources for leukocytes. The monocytes were analysed by flow cytometry in a mixed leukocyte suspensions obtained after ammonium chloride mediated lysis and in mononuclear cell suspension prepared by density gradient centrifugation (Ficoll-Paque) performed both at 4 degrees C and at 20 degrees C. Monocytes from heparinized blood had a higher expression of CD11b/CD18 and a more pronounced inter-individual variation than monocytes from EDTA blood. Monocytes isolated by Ficoll-Paque had a higher degree of ex vivo activation by means of altered expression of CD11b/CD18 and CD62L compared to monocytes prepared by ammonium chloride mediated lysis. This was more pronounced when the isolation procedure was performed at 20 degrees C.Our findings indicate that monocytes prepared by ammonium chloride mediated lysis of EDTA blood and with the cell handling temperature kept at 4 degrees C are exposed to the smallest ex vivo modulation by means of receptor alteration. An awareness of ex vivo modulation by different cell preparation procedures is of importance especially when comparing the expression of functional receptors on leukocytes of disparate origin.