免疫小鼠脾脏中抗原提呈细胞的快速分离

应用胶原酶消化脾脏、 Ｂ Ｓ Ａ 梯度离心富集低浮密度细胞和 Ｆ Ａ Ｃ Ｓ分选细胞法，自小鼠脾脏中分离出高纯度树突细胞、吞噬细胞和 Ｂ淋巴细胞。分离的 ３ 种细胞样品中分别含 ９７％ 树突细胞、９６％ 吞噬细胞和 ９９％ Ｂ淋巴细胞。而纯化前，树突细胞和吞噬细胞在低浮密度细胞中仅分别占 ６５％ 和 ３５％ ， Ｂ 淋巴细胞占脾细胞的 ４２％ 左右。该程序简便、快速、准确，在一个工作日内可制备大量细胞， 并适用于其他淋巴组织中抗原提呈细胞的纯化。

RAPID PURIFICATION OF ANTIGEN-PRESENTING CELLS FROM SPLEENS OF IMMUNIZED MICE

Professional antigen presenting cells (APCs) include dendritic cells (DCs), macrophages and B cells. A procedure for rapid isolation of these APCs from spleens of immunized mice was devised, involving collagenase digestion of tissues, enrichment of low buoyant cell fraction by dense BSA gradient centrifugation, and further purification by fluorescent activated cell sorting (FACS). Strikingly, the cell preparations contained 97% DCs, 96% macrophages and 99% B cells, respectively, compared to low levels of DCs (6.5%) and macrophages (3.5%) in low buoyant cell fraction, and less B cells (42%) in spleen cells. This procedure was simple, rapid, and able to prepare large numbers of APCs during one working day. It was also applicable to purification of APCs from other lymphoid tissues.