Isolation,purification and identification of mesenchymal stem cells derived from human umbilical cord

Aim:To isolate,purify and identify mesenchymal stem cells derived from human umbilical cord. Methods: Mesenchymal stem cells were isolated from human umbilical cord by digestion of collagenase Ⅳ and their unique growth characteristic of attaching to the wall of cell culture flask.The cell proliferation was analyzed by MTT assay and the growth curve was drawn.Their surface markers were detected by flow cytometry.The cells were induced to osteoblast with dexamethasone,antiscorbic acid and β-sodium glycerophosphate.And they also were induced to adipocytes with dexamethasone,insulin and indomethacin.After induction,the cells were observed by ALP staining and oil red O staining.The umbilical cord MSCs was injected into the BALB/c nude mice kidney's capsule to observe tumorigenicity.Results: Mesenchymal stem cells derived from human umbilical cord were spindle-shaped adherent cells.They have strong proliferation ability.They were strongly positive expression for CD44 and CD29,less expression for CD106,but negative expression for CD14,CD31,CD34,CD45 and HLA-DR.After induction,oil red O staining and ALP staining were positive.After transplantation,we find no tumorigenicity.Conclusion:The adherent cells isolated from from the umbilical cord have biological characteristics of mesenchymal stem cell.