Traditional breeding methods of hazelnut can not meet requirement of market. With the Corylus heterophylla Fisch.×C. avellana L. ‘Liaoning hazel No.7’, we study on the basic medium and the hormone combination in subculture using in vitro culture, in order to find a practical way of the propagation technology with hazelnut. The results showed that: (1) Compared to the medium composition and effect of seedling growth between DKW, NRM and WPM, the final selection of improved NRM medium as a subculture medium. (2) Choose 6-BA 2.5 mg/L and IBA 0.01 mg/L as the subculture hormones. The older generation to reduce its concentration to 6-BA 2.0 mg/L and IBA 0.005 mg/L that help to promote the proliferation of hazelnut seedlings. (3) We filter out the TDZ suitable concentration was 0.001 mg/L to 0.01 mg/L and the effect of the best was 0.005 mg/L. (4) The multiplier effect was not obvious for GA3. To add GA3 and polyamines at the same time was better than to add GA3 only and worse than a single addition of polyamines.