Cellular Differentiation of Human THP-1 Leukemia Cells with Phorbol-12-Myristate-13- Acetate (PMA)

Objectives: In this study, it was aimed to evaluate appropriate model of application time and dosage of PMA to get activated macrophages from monocytic cell line THP-1 cells.The differentiation of THP-1 cells into macrophages when stimulated with different dosage and application time of phorbol myristic acid (PMA) was shown morphologically and the effects on cell viability were evaluated.Materials and Methods: THP-1 monocytic cells (2x105 cells/ml) were treated with PMA of 10, 20, 40, 60 ng/ml concentrations and incubated for 24 h, 48 h and 72 h to induce differentiation into adherent macrophage-like cells.Differentiation was confirmed by observation of cell adherence and phenotypic changes using phase contrast microscopy.Cell viability was determined by trypan blue staining.Results: Results and conclusion: The best results were obtained with 20 ng/ml PMA at 48 h incubation and the cell viability was 92.2 % at this concentration.The administration of 40 ng/ml PMA resulted in a significant decrease in cell count compared to 10 and 20 ng/ml PMA administration at the 48h, but no difference was observed between groups after 72 h application.These results indicate that the differentiation of THP-1 monocytes to macrophages requires optimization to ensure that this invitro macrophage model more precisely reflects real physiologic conditions.These findings suggests that THP-1 cells are well differentiated by 20 ng/ml PMA at 48 h incubation.