Kumagai K, Itoh K, Hinuma S, Tada M
J Immunol Methods. 1979;29(1):17-25. doi: 10.1016/0022-1759(79)90121-2.
We have developed a simple method which can recover the highly purified macrophages or monocytes in suspension from mouse peritoneal exudate cells and human perpheral blood mononuclear cells. Plastic Petri dishes coated overnight with heat-inactivated fetal calf serum (FCS) selectively bind macrophages and monocytes. The adherent macrophages and monocytes are easily removed by incubation in phosphate-buffered saline containing 0.2% ethylenediamine tetraacetate (EDTA) and 5% FCS, and recovered as a cell suspension with greater than 95% purity. A small number of isolated cells can restore the mitogenic response to phytohemagglutinin (PHA-P) of macrophages-depleted lymphocytes and can lyse 51Cr-labeled target cells in an antibody-dependent cell-mediated cytotoxicity system. Thus, the method should be valuable for studies of various functions of macrophages and monocytes from different immune tissues of man and animals.
我们开发了一种简单的方法，该方法可以从小鼠腹腔渗出细胞和人外周血单核细胞中回收悬浮状态下高度纯化的巨噬细胞或单核细胞。用热灭活胎牛血清（FCS）包被过夜的塑料培养皿可选择性结合巨噬细胞和单核细胞。通过在含有0.2%乙二胺四乙酸（EDTA）和5% FCS的磷酸盐缓冲盐水中孵育，可轻松去除贴壁的巨噬细胞和单核细胞，并作为纯度大于95%的细胞悬液回收。少量分离的细胞可以恢复巨噬细胞缺失的淋巴细胞对植物血凝素（PHA-P）的促有丝分裂反应，并且可以在抗体依赖性细胞介导的细胞毒性系统中裂解51Cr标记的靶细胞。因此，该方法对于研究人和动物不同免疫组织中巨噬细胞和单核细胞的各种功能应该是有价值的。
