论文部分内容阅读
The aim of this study is to investigate the effect of electrical stimulation (ES) of varying pulse duration and frequency on proliferation and differentiation of C2C12 myogenic satellite cells in two-dimensional cultures.We stimulated C2C12 satellite cells with different combinations of pulse and duration and frequency, and dtermiend the optimal paradigm.The cellular properties were assayed using MTT vability assay, flow cytometry to determine cell cycle progression, and quantitative real-time PCR.With increasing duration of ES, we found a significant decrease in cell viability (P =0.000) and MyoD expression (P =0.018) , specifically in 20 ms long pulses, at 72 hours following electrical stimulation.Also, the frequency of pulses correlated significantly with the percentage of S-phase cells (P =0.004) and the expression of MyoD (P =0.003) and desmin (P =0.004), specifically at 80 pps stimulation.With increasing pulse frequency, we found an increase in MyHC2B expression (P =0.000).Expression of MyHC isoforms is regulated differently in response to variations in pulse duration versus frequency.Variations in duration and frequency of electrical stimulation affect proliferation and differentiation of C2C12 cells.We determined the optimal ES paradigm for myogenic proliferation and differentiation of cultured satellite cells.