Objective:To compare the functions of endothelial progenitor cells (EPCs) differentiated from cryopreserved and fresh bone marrow-derived mononuclear cells (MNCs). Methods: The bone marrow samples were taken from swine iliac bones. The isolated MNCs were cultured or cryopreserved at -80℃ for 3 months and then cultured again. The P1-EPCs were identifed by Dil-ac-LDL and FITC-UEA-1 double staining,immunohistochemistry and flow cytometry. The EPC pick-up rate,migration,adhesion, and proliferation abilities were compared between the cryopreserved group and the fresh group. Results: Immunohistochemisty showed that the P1-EPCs of the cryopreserved group were positive for CD133 (+),CD34 (+),CD31 () and KDR (); flow cytometry also showed they were positive for CD133 (\[17.24±3.12\]%),CD34 (\[37.21±10.85\]%),CD31 (\[72.07±13.34\]%) and KDR (\[89.09±16.40\]%). There were no significant differences in the pick-up rates (\[1.1±0.078\]% vs \[1.03±0.061\]%,P=0.054),migration rates (\[15±0.71\]% vs \[14.2±0.63\]%,P=0.17),adherence rates (\[42.7±2.1\]% vs \[39.5±1.7\]%,P=0.11),and proliferation abilities (\[25.06±2.82\]×104 vs \[21.64±2.34\]×104,P=0.089) between EPCs of the fresh and cryopreserved groups. Conclusion: Cryopreservation has no measurable influence on the numbers and functions of EPCs differentiated from bone marrow-derived MNCs,so cryopreservation can be used to obtain sufficient homogeneous EPCs in a short period for therapy using EPCs transplantation.