Abstract: Aseptic seeding and the establishment of regeneration system play an important role in crossbreeding of orchid, obtaining a certain number of hybrid progeny population, and providing basic materials for the breeding of new varieties. An aseptic seeding propagation for interspecific hybrid between Cymbidium hybridum‘Ken Ni’and C. sinense‘Pacific Ocean’was established. Different minimal medium, hormone and additives were designed to study the effects on the germination, rhizome proliferation and differentiation and seedling root. The results showed that 1/2MS 6-BA 1.5 mg/L NAA 0.5 mg/L AC 1.0 g/L was the suitable medium for the hybrid seed germination, 1/2MS 6-BA 2.0 mg/L NAA 0.5 mg/L was the ideal medium for the rhizome proliferation and differentiation, the differentiation rate was 88.6% after culturing for 70 days. MS 6- BA 1.0 mg/L NAA 2.0 mg/L banana 90 g/L AC 2.0 g/L was the right medium for inducing the root growth. Test- tube train seedling 30- 40 days in closed bottle and opening lid after 3- 5 days in greenhouse, fermentation bark and blinding were best media for temporary planting of tissue culture seedling. About 60 days, the seedling survival rate was 98.62%.