Co-culture of myoblasts with differentiated PC12 cells induced by nerve growth factor

Objective To explore the effect of neuron-like cells on proliferation and differentiation of C2C12 cells. Methods A co-culture system of PC12 cells and C2C12 cells was established using Transwell.They were allocated into 3 groups at random.In the blank control group,merely C2C12 cells were cultured; in the experimental group,the C2C12 cells were co-cultured with differentiated PC12 cells; in the negative control group,the C2C12 cells were co-cultured with undifferentiated PC1 2 cells.The effects of nerve growth factor (NGF) on differentiation of PC12 cells were identified by the immunofluorescence method.Proliferation of C2C12 cells was determined by flow cytometry.The expressions of Myogenin and Desmin in the C2C12 cells co-cultured with PC12 cells at 3 and 7 days were detected by RT-PCR and qPCR. Results The PC12 cells showed neuron-like appearance with neurite structure due to the effects of NGF and were MAP-2 positive.The differentiated PC12 cells inhibited the proliferation of C2C12 cells.The percentage of C2C12 cells at the DNA GI period in the experimental group (77.2％ ±0.4％ ) was significantly higher than that in the blank control group (71.0％ ± 0.6％ ) and in the negative control group (70.8％ ± 0.8％ ) ( P ＜0.05).The proliferation index (PrI) in the experimental group (22.8％ ± 0.4％ ) was significantly lower than that in the blank control group (29.0％ ± 0.6％ ) and in the negative control group (29.2％ ± 0.8％ ) ( P ＜0.05).The expressions of Myogenin and Desmin in the C2C12 cells in the experimental group were significantly higher than those in the other 2 groups at the same time point. Conclusion The differentiated PC12 cells may inhibit the proliferation of C2C12 cells but activate their differentiation. Key words: Myoblasts; Mice, inbred C3H; PC12 cells; Cell proliferation; Cell differentiation