Active Macrophage Inflammatory Protein 1 Alpha (MIP1a)

MIP-1a (macrophage inflammatory protein 1-alpha) also known as Chemokine (C-C motief) ligand 3 (CCL3), is a cytokine belonging to the CC chemokine family that is involved in the recruitment and activation of macrophages, monocytes and neutrophils. In this case, chemotaxis assay used 24-well microchemotaxis system was undertaken to evaluate the chemotactic effect of MIP-1a on the human monocytic cell line THP1. Briefly, THP1 cells were seeded into the upper chambers (100µl cell suspension, 106 cells/ml in RPMI 1640 with 0.5% FBS) and MIP-1a (100ng/mL, diluted in serum free RPMI 1640 ) was added in lower chamber with a polycarbonate filter (8µm pore size) used to separate the two compartments. After incubation at 37oC with 5% CO2 for 5h, the filter was removed, then cells in low chamber were observed by inverted microscope at low magnification (×40) and the number of migrated cells were counted at high magnification (×400) randomly (five fields for each filter).
By counting migrated cells in low chamber at high magnification (×400) randomly, it was shown that a mean of 41.2 THP1 cells/field migrated towards serum free RPMI 1640 medium with 100ng/mL MIP-1a, while only 3.6 THP1 cells/field migrated towards serum free RPMI 1640 medium. And the migrated THP1 cells in low chamber at low magnification (×40) was shown in Figure 1.