HL-60 differentiation in neutrophil-like cells

[itarille]

Hi all,

I would like to differentiate HL-60 cell line in neutrophil-like cells.
Looking at the protocols in the literature it seems very simple, but I don’t manage to have neutrophil-like cells.
I incubate the cells in RPMI 1640 medium complemented with 10% fetal bovine serum and 1.3% DMSO, for several days.
The problem it’s that they still continue to grow and the morphology doesn’t change really (they just begin a little bit smaller, but the nucleus is still big and round).
So, does someone can give me some advices or a protocol to help me please?
I also would like to know if I need to often change the medium with fresh DMSO or just to leave the cells until the differentiation occurs.

Thanks a lot for your help!