Mentzer S J, Guyre P M, Burakoff S J, Faller D V
Cell Immunol. 1986 Sep;101(2):312-9. doi: 10.1016/0008-8749(86)90144-9.
A previously unreported property of human mononuclear phagocytes is the ability of these cells to spontaneously aggregate. Fresh mononuclear cells obtained after plateletpheresis were noted to spontaneously form large cellular aggregates. Dual parameter immunofluorescence analysis demonstrated that the aggregating cells were positive for the monocyte marker CD11 (complement receptor, type 3) but were negative for the lymphocyte marker CD3 (T3 antigen). In addition, less than 5% of the nonaggregating cells were CD11+, suggesting that almost all CD11+ cells aggregated. Cellular aggregates were independent of cell concentration and formed more efficiently at 4 degrees C than at either 22 or 37 degrees C. Based on these observations, a purification procedure utilizing Ficoll-Hypaque separation, spontaneous aggregation at 4 degrees C, and transient plastic adherence resulted in a sevenfold enrichment of the CD11+ peripheral blood monocytes. Purified monocytes were contaminated with less than 2% CD3 cells. The size, growth, and adherence characteristics as well as cytologic stains indicated that the monocytes were not significantly altered by the purification procedure. Thus, spontaneous aggregation is an efficient and convenient method for the isolation of large numbers of purified monocytes.
人类单核吞噬细胞一个先前未被报道的特性是这些细胞能够自发聚集。在采集血小板后获得的新鲜单核细胞被观察到能自发形成大的细胞聚集体。双参数免疫荧光分析表明，聚集的细胞单核细胞标志物CD11（补体受体3型）呈阳性，但淋巴细胞标志物CD3（T3抗原）呈阴性。此外，不到5%的非聚集细胞为CD11+，这表明几乎所有CD11+细胞都发生了聚集。细胞聚集体与细胞浓度无关，在4℃时比在22℃或37℃时更有效地形成。基于这些观察结果，利用Ficoll-Hypaque分离、4℃下的自发聚集和短暂的塑料黏附的纯化程序，使CD11+外周血单核细胞富集了7倍。纯化的单核细胞被CD3细胞污染的比例低于2%。大小、生长和黏附特性以及细胞染色表明，纯化程序并未使单核细胞发生显著改变。因此，自发聚集是一种高效且便捷的方法，用于分离大量纯化的单核细胞。
