Semple J W, Allen D, Chang W, Castaldi P, Freedman J
Division of Hematology, St. Michael's Hospital, Toronto, Ontario, Canada.
Cytometry. 1993 Nov;14(8):955-60. doi: 10.1002/cyto.990140816.
Rapid purification of human lymphocyte subpopulations is an essential step in order to elucidate their interactions and/or contributions in various disease states. Cell purification using a Magnetic Activated Cell Sorter (MACS) is a relatively new technology which has been shown to be rapid and yield highly purified populations of cells. This report describes both a simple one-step positive selection method using the MACS to purify either human CD4+ or CD19+ lymphocytes from PBMC and a sequential separation of both CD4+ and CD19+ cell populations. These methods can separate the cell populations in approximately 4 h with yields > 90% and purity of 97 +/- 3% for CD4+ T cells and 92 +/- 5% for CD19+ B cells. In functional studies, purified CD19+ B cells secreted 13- and 24-fold more IgM and IgG, respectively, than the CD19- cell fraction in 10 day B cell stimulation assays. Purification of the two cell types did not cause any significant activation as shown by proliferation. Both cell types, however, were able to proliferate upon stimulation with interleukin-2.
快速纯化人淋巴细胞亚群是阐明它们在各种疾病状态下的相互作用和/或作用的关键步骤。使用磁性激活细胞分选仪(MACS)进行细胞纯化是一项相对较新的技术,已证明该技术速度快且能产生高度纯化的细胞群体。本报告描述了一种使用MACS从外周血单核细胞(PBMC)中纯化人CD4 +或CD19 +淋巴细胞的简单一步阳性选择方法,以及CD4 +和CD19 +细胞群体的顺序分离。这些方法可以在约4小时内分离细胞群体,CD4 + T细胞的产量> 90%,纯度为97 +/- 3%,CD19 + B细胞的纯度为92 +/- 5%。在功能研究中,在10天的B细胞刺激试验中,纯化的CD19 + B细胞分泌的IgM和IgG分别比CD19 -细胞部分多13倍和24倍。如增殖所示,两种细胞类型的纯化均未引起任何明显的激活。然而,两种细胞类型在用白细胞介素-2刺激后均能够增殖。