Sorting strategy and postsort evaluation of NA cells. After 7 days of culture, bulk DC-driven MLR cultures were stained with anti-CD25-PE and cell sorted. (A) A small cell gate was set on the forward versus side scatter 2-dimensional plot to select for resting cells, and from this population CFSE^bright and CD25^- cells were isolated. (B) Postsort evaluation of scatter profile and CFSE versus activation antigen. Cells were always more than 92% viable and 99% pure. Typically, purities ranged from 99.5% to 99.9% (8 donor combinations).