The vegetative and floral characteristics of chrysanthemum [Dendranthema grandiflora (Ramat.) Kitam.] plantlets that had survived after in vitro conservation at different periods and in different media were assessed after growing them in vivo. The surviving plantlets of varieties ‛Pasopati’, ‛Padma Buana’, ‛Puspita Nusantara’ and ‛Tirta Ayuni’ previously conserved under low-temperature conditions in Murashige and Skoog (MS) + 2.5% dimethyl sulfoxide (DMSO), full, ½ and ¼ strengths Tsuchiya media for 2, 4, and 6 mo were acclimatized under protected house conditions with standard cultural maintenance until flowering stage. The initial deviations in plant height, stem diameter, leaf length-width ratio and number of florets within certain culture media and within storage periods were not observed as the storage periods were increased. The size of the flowers, florets and discs, and the color of the adaxial and abaxial parts of all varieties were not affected by the conservation media and length of storage. Cytological alterations in terms of chromosome number, chromosome classification, and grouping were not found within variety and within media. The study showed that in vitro conservation of chrysanthemums is an effective alternative method to replace the more expensive and risky maintenance of live and field-grown germplasm without sacrificing genetic stability.