The Biological Characteristics and Culturing Conditions of Human Umbilical Cord Mesenchymal Stem Cells

Objective To optimize the isolation and culture conditions of mesenchymal stem cells(MSCs)in human umbilical cord in vitro,and explore their biological properties.Methods Human umbilical cord blood was collected in sterile condition,then the mononuclear cells were obtained by density gradient centrifugation and cultured in IMDM medium containing 10% fetal bovin serum.After the adherent mononuclear cells were obtained,the investigation and immunocytochemical identification of the cell cycles,cell growth,apoptosis of the mononuclear cells were conducted.Result MSCs obtained by Percoll(1.073g/ml)were similar in size,spindle-shaped or star-shaped epidermal-liked cells,expanded in size,and fibroblasts-like cells increased after passage culture.Cell growth curve analysis indicated that MSCs were in the exponential stage after 5d culture and in the stationary stages after 9d culture.FACS analysis showed that 14.8% of MSCs were in G2+S phase,the CD45 positive cells were about 42% by immunocytochemical identification.Conclusion The human umbilical cord derived mesenchymal stem cells were grown stably in vitro and can be used as the seed-cells in tissue engineering.