The Isolating,Cultivation and Identification of Human Umbilical Cord Blood Derived-mesenchymal Stem Cells

Objective Isolation and cultivation of mesenchymal stem cells(MSCs) in human umbilical cord in vitro,and determine their biological properties.Methods The mononuclear cells were isolated by density gradient centrifugation from human umbilical cord blood in sterile condition,and cultured in DMEM medium containing 10% fetal bovine serum.After the adherent mononuclear cells were obtained,the shape of cells were observed by microscope,then the cell growth curve,the cell cycle and the cell surface antigens were obtained by immunocytochemistry and flow cytometry methods.Results MSCs obtained by Percoll(1.073 g/mL) were similar in size,spindle-shaped or star-shaped fibroblasts-liked cells.Cell growth curve analysis indicated that MSCs were in the exponential stage after 5d and in the stationary stages after 9d.Flow cytometry analysis showed that the CD29 and CD44 positive cells were about 50%～70%.Conclusions The human umbilical cord derived mesenchymal stem cells were grown stably in vitro and can be used as the seed-cells in tissue engineering.