Methods for Obtaining Purified Lymphocytes, Glass-adherent Mononuclear Cells, and a Population Containing Both Cell Types From Human Peripheral Blood

Abstract

Methods are presented for obtaining simultaneously or separately two populations of cells from human peripheral blood, lymphocytes and monocytes, both of which are required to obtain blastogenesis and DNA synthesis in human leukocyte cultures. A simple 5-min centrifugation of heparinized blood yields a mononuclear leukocyte culture fluid containing 70-90% lymphocytes with few granulocytes but with sufficient numbers of monocytes so that the latter cell is not a limiting factor in the blastogenic reaction. A method is also presented for removing both granulocytes and monocytes from lymphocyte populations in a manner that permits monitoring and choice of the degree of lymphocyte purification. A method is also presented for obtaining glass-adherent mononuclear cells that do not undergo blastogenesis but will enable suitably stimulated "purified" lymphocytes to undergo blastogenesis. Studies of the function and morphology of these different cell populations are presented.