Isolation of peripheral blood mononuclear cells (PBMCs) is fraught with challenges including, but not limited to, the cost of limited gradients available for the isolation of PBMCs. Glycerol gradient (1.077 g/ml) was used to isolate PBMCs from adult peripheral blood. The differentiation potential of the isolated cells was assessed by culturing the cells in MEM at 37 °C in 5 % CO2. The results demonstrated that the isolated cells could differentiate into committed linages of the erythroid progeny. LDH assay revealed that glycerol was not cytotoxic to the cells. The use of glycerol density as an alternative could be significant in cell culture experiments.