Chestnut as an indigenous and valuable forest species exists in some forests of Guilan province, which the area of its stands is decreasing due to the lack of natural regeneration and socioeconomic problems as well as ink and blight diseases; therefore, chestnut’s asexual propagation through tissue culture methods can be effective in protecting this species and preventing its extinction. The aim of this study was to use the micropropagation method in order to produce complete chestnut plantlets by identifying the best explants and plant growth regulators. For this purpose, seeds were collected from Visroud site, sterilized, soaked in boiling water and cultured on B₅ medium, and then explants were taken from germinated seeds. Leaf, stem and root explants from in vitro seedlings of chestnut were cultured on B₅ containing cytokinins (TDZ and BA) with concentrations of 0.02, 0.2, 1 and 2 mg/l and auxins (IBA, NAA, and IAA) with concentrations of 0.1, 0.2, 1 and 2 mg/l. Results showed that the best response was obtained from the leaf explants and TDZ (0.2 mg/l) and IBA (2 mg/l) had the highest percentage of callus- shooting (32.4%) and rooting (80%), respectively. The survival of seedlings outside the laboratory was 80%. Considering the importance of chestnut species in economic and environmental terms, the present study can be useful for the implementation of reclamation management to protect, develop and prevent extinction of this species.