Richard, We routinely do live/dead separations of primed T cell cultures using histopaque/ficoll before restimulation for cytokine production. We've noticed no spontaneous cytokine production of 'ficolled' cells as measured by intracellular cytokine staining (IL-1, IFN-g, etc) - i.e., if the separated cells are cultured in monensin with no restimulus, no cells make cytokine. Andrew D. Wells, Ph.D. University of Pennsylvania Department of Medicine 728 Clinical Research Building 415 Curie Boulevard Philadelphia, PA 19104 (215) 573-1840 [office] (215) 898-1951 [laboratory] (215) 573-2880 [FAX] adwells@mail.med.upenn.edu
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