Abstract: Objective To explore isolating,culturing,identifying,freezing and thawing mesenchymal stem cells(MSCs)approach from Wharton j elly of human umbilical cord.Methods The MSCs were isolated from neonate umbilical cord,cultured by tissue explants adherent method.The surface markers were identified by flow cytometry.Passage cells,frozen 6 months, were thawed,then their biological characteristics were identified.Results MSCs were easily obtained from Wharton j elly of human umbilical cord via the proposed approach of tissue adherence.The flow cytometry analysis showed that MSCs expressed CD29,CD44,CD90 and CD105 positively,but negatively for CD34,CD45.The living cells of MSCs were 80%-90% after having been frozen and the thawed cells had the same characteristics as the previous.Conclusion MSCs can be successfully isolated from Wharton j elly of human umbilical cord by this method. The stem cells derived from Wharton j elly of human umbilical cord may be a novel alternative source of human MSCs for experimental and clinical applications.

Key words: umbilical cord, mesenchyma stem cells, fetal blood