从牛外周血单个核细胞中纯化和去除单核细胞的改进方法。单核细胞对凝集素反应的功能评估。

Improved methods for purification and depletion of monocytes from bovine peripheral blood mononuclear cells. Functional evaluation of monocytes in responses to lectins.

作者信息

Goddeeris B M, Baldwin C L, ole-MoiYoi O, Morrison W I

出版信息

J Immunol Methods. 1986 May 22;89(2):165-73. doi: 10.1016/0022-1759(86)90354-6.

Abstract

We have compared different techniques for the enrichment and depletion of monocytes from bovine peripheral blood mononuclear cells. Adherence to plasma-coated gelatin was the most efficient and reproducible method for enrichment of monocytes (80% monocytes), whereas depletion of peripheral blood mononuclear cells of monocytes (0.3% monocytes and less) was best achieved by defibrination of the blood from which the PBM were separated. In both instances, purity of the cell population could be improved further by an additional step, namely, FACS sorting with a monocyte-specific monoclonal antibody to purify monocytes (97% monocytes and more), and adherence to polystyrene to remove residual monocytes from defibrinated PBM (0.1% monocytes and less). Depletion of monocytes abolished the response of PBM to concanavalin A and phytohaemagglutinin. The lectin-induced response could be restored by adding gelatin/plasma purified monocytes. This activity of monocytes could be replaced by 2-mercaptoethanol.

摘要

我们比较了从牛外周血单个核细胞中富集和去除单核细胞的不同技术。贴壁于血浆包被的明胶是富集单核细胞（80%单核细胞）最有效且可重复的方法，而去除外周血单个核细胞中的单核细胞（0.3%及更少的单核细胞）最好的方法是对分离出外周血单个核细胞的血液进行去纤维蛋白处理。在这两种情况下，细胞群体的纯度都可以通过额外的步骤进一步提高，即使用单核细胞特异性单克隆抗体进行荧光激活细胞分选以纯化单核细胞（97%及更多的单核细胞），以及贴壁于聚苯乙烯以从去纤维蛋白的外周血单个核细胞中去除残留的单核细胞（0.1%及更少的单核细胞）。去除单核细胞消除了外周血单个核细胞对刀豆球蛋白A和植物血凝素的反应。通过添加明胶/血浆纯化的单核细胞可以恢复凝集素诱导的反应。单核细胞的这种活性可以被2-巯基乙醇替代。