Influence of NGF on PC12 cell neuronal differentiation activity

PC-12 is a classic neuronal cell line. Studies have suggested that NGF can induce PC-12 cell differentiation (26-28). Thus, undifferentiated PC12 cells (The Cell Bank of Type Culture Collection of the Chinese Academy of Sciences) were cultured in RPMI-1640 medium (cat. no. SH30809.018; Hyclone; Cytiva) supplemented with 10% horse serum (cat. no. 26050088; Gibco; Thermo Fisher Scientific, Inc.) and 5% FBS, and were treated with 10, 50 and 100 ng/ml NGF cell supernatants from NGF-overexpressing MDA-MB231 cells at 37˚C. NGF was obtained from the supernatants of NGF-overexpressing cells and the concentrations were confirmed by ELISA (Human NGF/NGF-β ELISA kit; cat. no. EK0469; Boster Biological Technology). NGF-β standards was used to establish the standard curve. The cell number and axon length were calculated by fluorescence microscopy (NIKON Ti-s inverted fluorescence microscope; Nikon Corporation); 250 cells from five fields of vision (n=50) were analyzed.