Haisch K, Gibbs B F, Körber H, Ernst M, Grage-Griebenow E, Schlaak M, Haas H
Division of Cellular Allergology, Research Center Borstel, Germany.
J Immunol Methods. 1999 Jun 24;226(1-2):129-37. doi: 10.1016/s0022-1759(99)00059-9.
Certain studies on basophils require highly purified functionally intact cell preparations. Here, a three-step procedure is described that meets these requirements. The procedure consists of a Ficoll density gradient step, counterflow elutriation and negative selection by magnetic cell sorting (MACS). The mean purity of basophils obtained from 30 donors was 97.6+/-3.96% with a viability of 99.6+/-0.83%. The recovery rate was 49.7+/-15.6%. The cells had a normal morphological appearance as assessed by May-Grünwald-stained cytospins and were functionally intact as shown by their unaltered capacity to release histamine and interleukin 4 (IL-4) following immunological activation. This procedure is a clear improvement over currently available techniques and should facilitate future investigations on basophils.
某些关于嗜碱性粒细胞的研究需要高度纯化且功能完整的细胞制剂。在此，描述了一种满足这些要求的三步程序。该程序包括一个Ficoll密度梯度步骤、逆流淘析以及通过磁性细胞分选（MACS）进行阴性选择。从30名供体获得的嗜碱性粒细胞的平均纯度为97.6±3.96%，活力为99.6±0.83%。回收率为49.7±15.6%。通过May-Grünwald染色的细胞涂片评估，细胞具有正常的形态外观，并且在免疫激活后释放组胺和白细胞介素4（IL-4）的能力未改变，表明其功能完整。该程序明显优于目前可用的技术，应有助于未来对嗜碱性粒细胞的研究。
