Wanda P E, Smith J D
J Histochem Cytochem. 1982 Dec;30(12):1297-300. doi: 10.1177/30.12.7153501.
A simple and general labeling method has been adapted for the rapid detection and isolation of heterokaryons soon after somatic cell fusion. The method involves prelabeling each parental cell type by adding one of two hydrophobic fluorescent probes, F18 or R16, to the culture medium. These probes are nontoxic, nonmutagenic, and do not inhibit either cellular replication or the efficiency of fusion. Following polyethylene glycol (PEG)-induced fusion, heterokaryons are identified on a cell sorter as cells showing fluorescence as a result of nonradiative resonance energy transfer (RET) between the two fluorochromes. Optimal conditions are described for the unambiguous detection of heterokaryons by fusion-induced RET. The value of this method lies in the universal applicability of F18 and R16 as simple and direct membrane labels for any pair of parental cells, even those lacking selectable genetic markers or detectable antigenic differences. This potential for heterokaryon selection in any cell cross should significantly expand the range of cell types amenable to investigation through somatic cell genetics, while the rapidity of the method should facilitate the study of early events following fusion.
一种简单通用的标记方法已被采用，用于在体细胞融合后不久快速检测和分离异核体。该方法包括通过向培养基中添加两种疏水性荧光探针之一F18或R16，对每种亲代细胞类型进行预标记。这些探针无毒、无致突变性，且不抑制细胞复制或融合效率。在聚乙二醇（PEG）诱导融合后，异核体在细胞分选仪上被识别为由于两种荧光染料之间的非辐射共振能量转移（RET）而显示荧光的细胞。描述了通过融合诱导的RET明确检测异核体的最佳条件。该方法的价值在于F18和R16作为任何一对亲代细胞的简单直接膜标记物具有普遍适用性，即使是那些缺乏可选择遗传标记或可检测抗原差异的细胞。这种在任何细胞杂交中选择异核体的潜力应能显著扩大适合通过体细胞遗传学进行研究的细胞类型范围，而该方法的快速性应有助于对融合后早期事件的研究。
