The aim of this study was to evaluate whether vitrification is an effective cryopreservation method for storage of human umbilical cord mesenchymal cells (HUCMs). HUCMs were vitrified using a two step method. The viability of vitrified HUCMs (v-HUCM) and non-vitrified HUCMs (n-HUCM) was determined by Trypan Blue staining. The expression of several markers was evaluated using flow cytometry. The osteogenic, adipogenic and chondrogenic differentiation potential of v-HUCMs and n-HUCMs was determined. The post-warming viability of HUCMs was 95.54 +/- 2.30. The expression of surface antigens (strong positive for CD44, CD90 and CD105; negative for CD34 and CD45) was similar in the above mentioned groups. The v-HUCM cells retained ability to differentiate into osteoblasts, adipocytes and chondrocytes under appropriate culture conditions. The analysis of these results showed that vitrification is a reliable and effective method for cryopreservation of human umbilical cord mesenchymal cells.