Separation of spontaneously differentiating and cell cycle-specific populations of HL-60 cells

Abstract

The human leukemia cell line HL-60 consists predominantly of abnormal promyelocytes. When grown in RPMI-1640 and 10% FCS between 5 and 10% of these cells spontaneously differentiate into more mature myeloid cells, becoming smaller in size and developing the ability to generate superoxide. Centrifugal elutriation was used to separate these G0 cells from the bulk of the cycling G1, S and G2M cells. These isolated differentiating cells are shown to be similar in size, DNA content, RNA content and NBT positivity not only to granulocyte induced HL-60 cells but also to human peripheral blood granulocytes. This methodology allows the study of differentiative vs proliferative processes through the quick one-step generation of homogeneous subpopulations of G0, G1, S and G2M cells.