Grisham M B, Engerson T D, McCord J M, Jones H P
J Immunol Methods. 1985 Oct 10;82(2):315-20. doi: 10.1016/0022-1759(85)90363-1.
Several different methods are currently used by numerous laboratories for the isolation of human neutrophils. These methods include partial purification by dextran sedimentation followed by water lysis, and more complete purification procedures utilizing discontinuous density gradients coupled with dextran sedimentation and in some cases hypotonic lysis. Some investigators refrain from using certain purification schemes because certain steps or reagents used in a particular method might adversely affect the functional parameter of the neutrophil they wish to measure. In spite of these concerns there has been no systematic comparison of the functional status of neutrophils prepared by the various methodologies. In this study we have compared 4 commonly used methods of isolation with neutrophil function. The results of this study indicate that while neutrophil yield and purity were determined by isolation procedure, all cells were equivalent with regard to chemotactic performance, ability to degranulate, and ability to produce superoxide and hypochlorous acid.
目前众多实验室采用几种不同的方法来分离人中性粒细胞。这些方法包括通过葡聚糖沉降进行部分纯化，随后进行水裂解，以及更完整的纯化程序，利用不连续密度梯度结合葡聚糖沉降，在某些情况下还包括低渗裂解。一些研究人员避免使用某些纯化方案，因为特定方法中使用的某些步骤或试剂可能会对他们想要测量的中性粒细胞的功能参数产生不利影响。尽管存在这些担忧，但尚未对通过各种方法制备的中性粒细胞的功能状态进行系统比较。在本研究中，我们比较了4种常用的分离方法与中性粒细胞功能的关系。这项研究的结果表明，虽然中性粒细胞的产量和纯度由分离程序决定，但所有细胞在趋化性能、脱颗粒能力以及产生超氧化物和次氯酸的能力方面是等效的。
