Protoplasts were isolated from the embryogenic calli of satsuma mandarin (Citrus unshiu Marc.) cv. Tokumori-Wase and cultured in the Gellan Gum-embedded Murashige and Tucker medium containing 40 mg/l adenine, 0.15M sucrose and 0.45M mannitol. The percentage of colony formation (plating efficiency) in 40 days of culture was 46%, which was much higher than that in MT medium without adenine. Most of the colonies grew to the size of about 1 mm in 2 months of culture and they were successfully transferred onto the MT medium containing 5% lactose for embryoid formation. Five percent of these embryoids germinated normally and grew to plantlets in 30-60 days after transplanting to the MT medium containing 1 mg/l gibberellic acid (GA3), 3% sucrose and 0.2% Gellan Gum. Protoplasts obtained in the culture of other two selections, Miyagawa-Wase and Okitsu-Wase also regenerated plants by using the same method as above.