Callus culture initiation of strawberry (Fragaria${ imes}$ananassa Duch.) was investigated at different Murashige and Skoog (MS) medium strengths, types and concentrations of plant growth regulators, and incorporating a cold pretreatment period to determine the optimal nutritional and environmental conditions. No high quality callus was induced on MS media without auxin regardless of medium strength. When 6-benzylaminopurine (BA) was combined with indole acetic acid (IAA), naphthalene acetic acid (NAA), and 2,4-dichlorophenoxyacetic acid (2,4-D), high quality callus were highly induced compared to medium supplemented with auxin alone. When $0.5mg{cdot}L^{-1}$ BA was combined with IAA, NAA, and 2,4-D, high quality callus induction was more effective than the medium supplemented with the other BA concentrations. The best combination of auxin and cytokinin for high quality callus induction was $1.0mg{cdot}L^{-1}$ NAA and $0.5mg{cdot}L^{-1}$ BA. Although the differences in callus induction were not significant, high quality callus induction at half strength MS medium was more effective than at full strength medium. When $30g{cdot}L^{-1}$ sucrose was added to the half strength MS medium, the rate of high quality callus induction increased. The optimum cold pretreatment temperature and period for high quality callus induction were $4^{circ}C$ and 72 h, respectively. Regeneration rate of high quality callus increased in MS medium supplemented with thidiazuron.