Zborowski M, Sun L, Moore L R, Chalmers J J
Department of Biomedical Engineering, The Cleveland Clinic Foundation, OH 44195-5254, USA.
ASAIO J. 1999 May-Jun;45(3):127-30. doi: 10.1097/00002480-199905000-00004.
Rapid and efficient cell sorting methods are important for tissue progenitor cell isolation. We built and evaluated a laboratory prototype of a continuous flow, quadrupole magnetic cell sorter. The sorter was tested on a model cell system of human peripheral lymphocytes. The helper T cell subpopulation was targeted by primary, mouse anti-CD4 monoclonal antibody conjugated to a fluorochrome (FITC), and magnetized by secondary, anti-FITC antibody magnetic colloid. The purities and recoveries of the cell fractions were measured by flow cytometry and an automated cell counter. Cells were spread across the flow according to their magnetophoretic mobilities. The purity of the CD4 cell enriched fraction was 99.6%, and the purity of the CD4 cell depleted fraction was 2% for an initial CD4 cell purity of 36%; the corresponding recovery of the enriched CD4 cell fraction was 59% at a sorting speed of 4,200 cells/s (four experiments). The recovery could be increased to 90% with a concomitant decrease in the purity of CD4 cell enriched fraction to 66%. This type of sorting should be applicable to any cells in suspension for which a suitable antibody exists, in particular, to large, fragile cells.
快速高效的细胞分选方法对于组织祖细胞的分离至关重要。我们构建并评估了一种连续流动四极磁细胞分选仪的实验室原型。该分选仪在人外周血淋巴细胞的模型细胞系统上进行了测试。辅助性T细胞亚群通过与荧光染料（异硫氰酸荧光素）偶联的一级小鼠抗CD4单克隆抗体作为靶点，并通过二级抗异硫氰酸荧光素抗体磁性胶体进行磁化。细胞组分的纯度和回收率通过流式细胞术和自动细胞计数器进行测量。细胞根据其磁泳迁移率在流动中分散。对于初始CD4细胞纯度为36%的情况，富集的CD4细胞组分的纯度为99.6%，耗尽CD4细胞的组分的纯度为2%；在分选速度为4200个细胞/秒时，富集的CD4细胞组分的相应回收率为59%（四个实验）。回收率可提高到90%，同时富集的CD4细胞组分的纯度降至66%。这种分选类型应适用于存在合适抗体的任何悬浮细胞，特别是大的、脆弱的细胞。
