A protoplast to plant regeneration system will open the way for somatic hybridization in cyclamen (Cyclamen persicum Mill.). A protocol for efficient plant regeneration from protoplasts isolated from embryogenic suspensions has been developed in cyclamen for the first time, and then successfully applied to different genotypes. Enzymatic digestion in 2.0% cellulase R10 and 0.5% macerozyme R10 resulted in yields between 1.0 and 5.0 x 10⁵ protoplasts per gram fresh mass depending on the genotype. Protoplasts were immobilized in alginate films, which promoted proper cell wall regeneration. Cell division was recorded for all eleven tested genotypes in frequencies of up to 12% and 18% after seven and 14 days, respectively. Plant regeneration via somatic embryogenesis succeeded in nine out of the 11 genotypes under investigation. Protoplast-derived plants from eight genotypes have been successfully transferred to soil. Somaclonal variation was detected in terms of ploidy levels of regenerants, malformed flowers, and formation of multiple vegetative meristems with delayed flowering. However, its degree depended on the genotype and the time period between isolation of protoplasts and plant regeneration.