武漢云克隆科技股份有限公司

PDGFA (Platelet-derived growth factor subunit A) is a Growth factor that plays an essential role in the regulation of embryonic development, cell proliferation, cell migration, survival and chemotaxis. PDGFA has been described as a chemoattractant for monocytes and proven to be able to induce chemotactic migration of THP-1 cells. Therefore, chemotaxis?assay used 24-well microchemotaxis system was undertaken to detect the chemotactic effect of PDGFA on the human monocytic cell line THP-1. Briefly, THP-1 cells were seeded into the upper chambers (100uL cell suspension,106cells/mL in RPMI 1640 with 0.5% FBS) and PDGFA (1.5ng/mL, 7.5ng/mL and 15ng/mL diluted separately 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 3h, 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). Result shows PDGFA is able to induce migration of THP-1 cells. The migrated THP-1 cells in low chamber at low magnification (×40) were shown in Figure 1. Five fields of each chamber were randomly chosen, and the migrated cells were counted at high magnification (×400). Statistical results?were shown in Figure 2. The optimum chemotaxis of PDGFA occurs at 1.5ng/mL.