Sepax gives an increased recovery of nucleated cells than all other methods. (A) Sepax offers the highest recovery of CD45+ cells at 75.79% (SD±18.58) compared to PrepaCyte CB at 72.03% (SD±8.48), Plasma depletion at 71.38% (SD±29.35), Hetastarch at 44.94% (SD±20.06), and Density Gradient at 19.13% (SD±10.88). Sepax was only significantly more efficient than Hetastarch and density gradient methods (p<0.005). (B) After exclusion of granulocytes recovery with Sepax is still the greatest at 78.8% (SD±21.36) compared to other methods. These results were determined using differential cell counts coupled with flow cytometric analysis of CD45+ cells, viability of CD45+ cells. Exclusion of granulocytes was also determined in this way to give a more accurate picture of cell recovery. Fluorochromes used were PE Cy5 for CD45+ cells, PE for CD14 +/− cells and 7AAD for viability.