Study on Isolating and Culturing Human Umbilical Cord Derived Mesenchymal Stem Cells in Vitro and its Biological Characteristics

Objective To establish a method to separate mesenchymal stem cells from human umbilical cord(UC-MSCs),culture UC-MSCs in vitro,and study its biological characteristics.Methods MSCs were separated from human umbilical cord with two enzyme method and expanding cultured in vitro.Growth curves of cells were drawn and surface antigens were detected with flow cytometry.tumorigeicities of cells were tested by soft agar cloning method.Results The methods of culture and expansion of UC-M SCs in vitro were established.Adhesive cells were all positive for MSC-related antigens,such as CD29,CD105,CD44,and CD13,but negative for CD34,CD45,CD31,CD106,and HLA-DR.Multipication of cells was 30 hours.Cell cycle showed that there was percentage of stem cells as G0 /G78.84% and S+G2+M11.16% respectively cells. The number of UC-MSCs didn't form clone in soft agar.Conclusion Two enzyme method is a good method to isolate and culture UC-MSCs.The cultured cells,with the biological characteristics of MSCs other than tumorigenic ability,can be amplified easily and passaged in vitro,providing a basis for MSCs banks and clinical application.