Isolation and identification of mesenchymal stem cells from human umbilical cord blood

Objective To establish the method of isolation and culture of mesenchymal stem cells(MSCs) from human umbilical cord blood(UCB) and to investigate whether its immunophenotype is coincidence with bone marrow-derived MSCs.Methods Human UCB sample was harvested from full term delivery by cesarean section.The mononuclear cells were separated from UCB using lymphocyted separation medium and suspended ones were removed in the later culture.The adherent cells′ proliferation and cell cycle were observed and their immunophenotyes were analyzed by flow cytometry(FCM).Results In low-glucose DMEM,containing fetal bovine serum,adherent cells exhibited mesenchymal-like phenotype.These cells were able to be purified and expanded by three passage.The population-doubling time of cells was approximately 60h.FCM showed that 85% of cells were in the phase of G0/G1 and these cells expressed CD29,CD44,CD54 and CD105,but no hematopoietic lineage markers,such as CD34 and CD45.Conclusion Non-hematopoietic stem cells can be isolated in UCB and they have the same immunophenotype with bone marrow-derived MSCs.