Among the species of the genus Anemone, A. coronaria L. is one of the most widely grown for cut flower production. As the homogeneity of F₁ seed population is still a problem, a study on the possibility of establishing a reliable system of in vitro multiplication of parental plants has been carried out. Seeds and immature flowers were used as explants. Multiplication phase was established using the cytokinin 2iP (9.84 µM) which increased both the multiplication rate and the fresh weight of the clusters. The rooting phase was studied by evaluating root development and the tuber enlargement. The rooting medium, containing 2.85 µM IAA and 2.43 µM IBA, allowed root emission and the increase of the tuber size during the four months of culture. After the acclimatization trials it was possible to identify a tuber weight threshold (500 mg) above which the acclimatization was good. The plants coming from in vitro culture grew normally. No phenotypic variants were noted and the quality of the flowers was comparable to the commercial standards.