In order to study multiplication culture of Tinospora sinensis, using the robust and pest-free young spears and leaves of the Tinospora sinensis as the research material, this article was to examine the disinfection methods and to study the tissue culture of the Tinospora sinensis through cultivating them in the hormone mediums of different concentrations. According to the results, if the spear surface was disinfected with the alcohol of 70% for 30 seconds, then soaked in the mercury of 0.1% for 5 minutes, rinsed with the sterile water twice, then soaked in the mercury of 0.1% for 5 minutes, rinsed with the sterile water 4 times, the outcome would be the best that the survival rate was 66.7%; while the leaves can be disinfected effectively with mercury for 12 minutes as the survival rate was 53.3%; the medium suitable for the bud induction and proliferation was MS+6-BA 0.05 mg/L+GA3 0.04 mg/L+NAA 0.02 mg/L, the multiplication coefficient for 30 days was 3; the medium suitable for callus induction was MS+2,4-D 1.5 mg/L+BA 0.5 mg/L, the spear induction rate can be as high as 83.3%, and the leave induction rate can be 76.7%, but the inducted callus has low differentiation, unable to differentiate adventitious buds; the medium suitable for root taking was MS+ NAA 0.1 mg/L, the root taking rate can be 100% and the transplant survival rate can be 96%.