Wright W E
Somatic Cell Genet. 1981 Nov;7(6):769-75. doi: 10.1007/BF01538763.
The selection of heterokaryons using irreversible biochemical inhibitors provides an effective method for isolating fusion products between any types of cultured cells. One weakness of the technique is that the surviving heterokaryons can only be obtained at low culture densities, making their analysis cumbersome. This report describes the use of Ficoll-sodium diatrizoate gradients to partially separate viable from nonviable cells and characterizes the effects of initial plating density and medium volume on heterokaryon survival. The combined result of optimizing these parameters increases cell rescue and permits heterokaryons to attach at densities of 60,000 cells/cm 2 instead of 600 cells/cm 2. These modifications substantially improve the ease of using irreversible biochemical inhibitors to isolate purified populations of heterokaryons.
使用不可逆生化抑制剂选择异核体为分离任何类型培养细胞之间的融合产物提供了一种有效方法。该技术的一个弱点是，只能在低密度培养条件下获得存活的异核体，这使得对它们的分析变得繁琐。本报告描述了使用聚蔗糖-泛影酸钠梯度从非存活细胞中部分分离出存活细胞，并表征了初始接种密度和培养基体积对异核体存活的影响。优化这些参数的综合结果提高了细胞存活率，并使异核体能够在60,000个细胞/cm²而不是600个细胞/cm²的密度下附着。这些改进极大地提高了使用不可逆生化抑制剂分离纯化异核体群体的便利性。
