Janssen J T, De Pauw B E, Smeulders J B, Wessels J M, Haanen C
J Immunol Methods. 1984 May 11;70(1):23-30. doi: 10.1016/0022-1759(84)90385-5.
Studies on splenic lymphocytes have hitherto been performed on single cell suspensions depleted of phagocytic cells by adherence to plastic or incubation with carbonyl iron. These techniques have the disadvantages of selective cell loss, suboptimal cell purification and cell activation. This paper describes purification of splenic lymphocytes by the use of counterflow centrifugation (CFC). The method was adapted to overcome pelleting of cells in the separation chamber to form a plug at the inlet and impede adequate flow. By combining 2 different separation chambers on line in 1 rotor this problem was overcome. Of all lymphocytes recovered after CFC 88.8 +/- 1.4% were collected in 2 pooled fractions with a purity of greater than or equal to 98% and a cell viability of 95%. After CFC, 80.8 +/- 12.1% of the viable cells loaded were recovered.
迄今为止，对脾淋巴细胞的研究是在通过贴壁于塑料或与羰基铁孵育而去除吞噬细胞的单细胞悬液上进行的。这些技术存在选择性细胞丢失、细胞纯化不理想和细胞活化的缺点。本文描述了使用逆流离心法（CFC）纯化脾淋巴细胞。该方法经过改进以克服细胞在分离室中沉淀，在入口处形成堵塞并阻碍充分流动的问题。通过在一个转子中在线组合两个不同的分离室，这个问题得到了克服。在CFC后回收的所有淋巴细胞中，88.8±1.4%收集在两个合并的组分中，纯度大于或等于98%，细胞活力为95%。CFC后，回收了80.8±12.1%的加载活细胞。
