Pick E, Kotkes P
J Immunol Methods. 1977;14(2):141-6. doi: 10.1016/0022-1759(77)90004-7.
A simple method for preparing culture supernatants containing macrophage migration inhibitory factor (MIF) by pulse exposure to guinea pig lymphocytes to concanavalin A (ConA), is described. The method is based on the property of originally nonadherent lymphocytes to attach to the flask surface in the presence of Con A. The Con A-induced lymphocyte 'monolayer' can be easily rinsed, allowing the removal of free Con A without repeated, cell damaging centrifugation. The Con A-pulsed lymphocytes are cultured for 24 h in medium and supernatants containing MIF but free of undesired Con A are regularly obtained.
本文描述了一种通过将豚鼠淋巴细胞短暂暴露于刀豆蛋白A(ConA)来制备含有巨噬细胞移动抑制因子(MIF)的培养上清液的简单方法。该方法基于原本不贴壁的淋巴细胞在ConA存在下附着于培养瓶表面的特性。ConA诱导的淋巴细胞“单层”可以很容易地冲洗,无需反复进行对细胞有损伤的离心操作即可去除游离的ConA。将ConA脉冲处理的淋巴细胞在培养基中培养24小时,可定期获得含有MIF但不含不需要的ConA的上清液。