Neutrophil granulocytes are the first line of defence in innate immunity, responding to invading pathogens and injuries. As neutrophil model for in vitro studies, HL-60 a promyelocytic leukemia cell line is often used. Upon induction with retinoic acid the cells differentiate into mature neutrophils. Retinoic acid, a physiological metabolite of vitamin A, plays a role in neutrophil maturation. Peripheral blood neutrophils can be used as in vitro model as well. Human serum albumin is able to bind endotoxin and influence the innate immunity. Our aim was the establishment of a neutrophil model to further investigate the effect of albumin species in combination with endotoxin on neutrophils. Cell morphology and surface antigen patterns were used to assess differentiation status of HL-60 and compared to isolated neutrophils. Proliferation rate evaluated HL-60 differentiation additionally. Oxidative burst and phagocytosis measured by flow cytometry assessed the functionality of differentiated HL-60. Albumin was isolated from serum with an exchange column. The oxidation and reduction of isolated albumin was done with H2O2 and GSH and assessed by HPLC. HL-60 showed signs of differentiation by stopping proliferation, segmentation of the nucleus and up-regulation of CD11b, but cells lack in phagocytosis and oxidative burst is impaired when stimulated with E.coli. Neutrophil isolations gain pure populations and negative selection allows the removal of residual cells from the population. To conclude, HL-60 cannot be used as in vitro model until cells show functional activity. The isolation of primary neutrophils is established and experiments with this model can start. Albumin isolation works well but an upscaling has to be done to yield higher amounts and the oxidation has to be further optimized.