Berkow R L, Tzeng D Y, Williams L V, Baehner R L
J Lab Clin Med. 1983 Nov;102(5):732-42.
Standard isolation techniques for the human polymorphonuclear leukocyte (PMN) involve sequential exposure of cells to the nonphysiologic environments of dextran, Ficoll-Hypaque (FH) gradient centrifugation, and hypotonic conditions. It has been suggested that these may be harmful to the recovered PMN. Counterflow centrifugal elutriation (CCE) allows separation of human PMNs while the cells are continuously bathed in a physiologic and isotonic buffer. To investigate whether preparative technique may alter PMN activation, we compared PMNs obtained by these two methods for stimulus-induced superoxide production and release of primary and specific granule contents. Resting PMN volume was also evaluated. We observed that PMNs obtained using the CCE method were larger and released significantly more superoxide and specific granule contents than PMNs obtained by the standard FH technique. The possible origins for these differences are discussed.
人类多形核白细胞（PMN）的标准分离技术包括将细胞依次暴露于葡聚糖的非生理环境、Ficoll-Hypaque（FH）梯度离心以及低渗条件下。有人认为这些可能对回收的PMN有害。逆流离心淘析（CCE）可在细胞持续浸泡于生理等渗缓冲液的同时分离人类PMN。为研究制备技术是否会改变PMN的激活，我们比较了通过这两种方法获得的PMN在刺激诱导下的超氧化物产生以及初级和特异性颗粒内容物的释放情况。还评估了静息PMN的体积。我们观察到，使用CCE方法获得的PMN比通过标准FH技术获得的PMN更大，并且释放出显著更多的超氧化物和特异性颗粒内容物。文中讨论了这些差异可能的来源。
