Abstract: Objective: To explore retroviral mediated neo gene transduction conditions and methods in primary myoblast of rhesus monkey Methods: To calculate the number of primary myoblast, after co cultured in vitro with supernatant of retroviral vector; To analyze phenotypes Thy-1, NCAM, Desmin of myoblast during transduction by using immunofluorescent staining and flow cytometry; Clonal PCR was employed for analysis of transduction efficacy and assessing stability of integrat ed neo gene Results: Highly purified myoblast Thy-1 （85.0± 5.0)%, NCAM （96.7±5.8)%, Desmin （95.7±2.1%) was obtained in F10 media supplemented with 20% FCS, 5% chick embryo extract No negative effect of supernatant of retroviral vector on these phenotypes of myoblast was observed The proliferation of myoblast was inhibited by supernatant of retroviral vector in QD24 （daily exposure to vector for 24 hours), but no negative effect was observed in Q D6（daily exposure to vector for 6 hours) In QD6, we not only obtained efficacy of transduction （63 9±7 3)%, but also observed no decreasing in efficacy of transduction 2 months post transduction Conclusion: These results present high-efficacy retroviral transduction and stability of integrated neo gene in primary myoblast of rhesus monkey, and can be used in investigating stem cell plasticity of muscle in large animal