PLGA nanoparticles (PLGA-NPs) are being extensively studied as drug carriers for their controlled release, biodegradability and biocompatibility. This study evaluated the cellular uptake of PEGylated PLGA-NPs in Hela cells. MePEG-PLGA (5%-15%) was used to prepare PEG modified PLGA nanoparticles (PEG-PLGA-NPs), and the fluorescent marker DiI was encapsulated in the nanoparticles for the visualized analysis. The nanoparticles were characterized for surface morphology, particle size, zeta potential, and for cellular uptake by Hela cells. Results showed that PLGA nanoparticles were lowly cytotoxic and could be uptaken by Hela cells freely. PEG-PLGA-NPs had faster cellular uptake than that of nude PLGA nanoparticles, especially 10%PEG-PLGA-NPs. It suggested that the surface modification of PLGA-NPs by PEG notably improved the cellular uptake.