Vitrification has been suggested to be an effective method for the cryopreservation of human ES cells. However, the efficiency of vitrification with different vehicles remains a matter of ongoing controversy. The objective of this study was to assess the efficiency of cryopreservation in human ES cells by vitrification using different vehicles. A human ES cell line and a variety of vehicles, including microdroplet (MD), openpulled straw (OPS) and electron microscopic grid (EMgrid), were employed in an attempt to assess vitrification efficiency. In order to evaluate the survivability and the undifferentiated state of the postvitrified human ES cells, we conducted alkaline phosphatase staining and characterization via both RTPCR and immunofluorescence assays. The survival rates of the postvitrified human ES cells using MD, OPS and EMgrid were determined to be 61.5%, 66.6% and 53.8%, respectively. There also exist significant differences between slowfreezing and vitrification (p<0.01). However, no significant differences were detected between the vehicle types. Finally, the pluripotency of human ES cells after thawing was verified by teratoma formation. Cryopreservation using vitrification is more effective than slowfreezing, and the efficiency of vehicles proved effective with regard to the preservation of human ES cells.