Isolation and identification of mesenchymal stem cells from human umbilical cord Wharton's Jelly

【Objectives】To explore methods for isolation of mesenchymal stem cells derived from human umbilical cord wharton's jelly and to identify the isolated cells which are able to differentiate into osteogenic and adipogenic tissues.【Methods】After removal of umbilical arteries,vein and tunica externa of umbilical cord,the remaining tissue was wharton's jelly.To obtain adherent cells,wharton's jelly of umbilical cord was cut into pieces and digested with 0.2% collagenase of type Ⅱ.The mesenchymal stem cells of umbilical cord were passaged.Phenotypes were detected with flow cytometry with surface antigens CD44,CD90,CD105,CD73,CD34,CD45,HLA-DR,HLA-ABC and osteogenic and adipogenic tissues were identified by differentiation.【Results】Wharton's jelly of human umbilical cord had abundant stem cells.The isolation of these cells had been proved easily when collagenase was used to detach the cells from Wharton's jelly.Flow cytometry analyses revealed that the stem cells from Wharton's jelly represented a nonhematopoietic cell popution.However they expressed amounts of MSCs markers,such as CD44,CD90,CD105,CD73,meanwhile positive expression of HLA-ABC was found in the stem cells,but not CD34,CD45,HLA-DR.Immunohistochemistry showed that the MSCs strongly expressed alkaline phosphatase and Von Kossa positive.【Conclusions】Mesenchymal stem cells can be successfully isolated from human umbilical cord Wharton's jelly.The stem cells derived from Wharton's jelly of human umbilical cord may be a novel alternative source of human MSCs for experimental and clinical applications.