A preliminary study on the interaction between Asn-Gly-Arg (NGR)-modified multifunctional nanoparticles and vascular epithelial cells

Liu, Chunxi; Liu, Tingxian; Yu, Xiaoyue; Gu, Yizhu

HUVEC were cultured in ECM with 1% ECGS, 5% FBS and 1% P/S solution in a 37 1C incubator with 5% CO 2 . The cells were detached by 0.025% trypsin/EDTA, and the cell density was adjusted to 1 Â 10 6 /100 μL PBS to grow to logarithmic phase. Then, cells were incubated with 2 μL of a PE-conjugated monoclonal antibody specific for CD13 to detect CD13 expression or DyLight 488-labelled anti-human CAV1 monoclonal antibody (1:200 dilution) to detect CAV1 expression for 1 h at 4 1C. After being washed in cold PBS for three times, the cells were resuspended in PBS and analyzed on FACS (BD Biosciences, USA) equipped with a 488-nm argon laser for excitation. For each sample, 10,000 events were collected, and fluorescence was detected. Signals were amplified in the logarithmic mode for fluorescence to determine the positive events by a standard gating technique. The percentage of positive events was calculated as the events within the gate divided by the total number of events, excluding cell debris.