Isolation of Peripheral Blood Mononuclear Cells Using Glycerol Density Gradient

Abstract

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.

In Vitro Cytotoxicity and Hemolysis Effect of Poly-Gamma-Glutamic Acid Nano-Polymer Biosynthesized Using Some Isolates of Bacillus spp.

“…The glycerol gradient was carried out by adding 1.68 mL into 25 mL of sterile water. NaCl (1.2) was added, making up 10% of the total volume [38]. The integrity of the isolated cells necessitates a gradient isotonic with the intracellular environment which in turn is necessary for the cell isolations [39].…”