Culture and identification of mesenchymal stem cells derived from human umbilical blood

Objective To establish the method of in vitro culture and expansion of human umbilical blood-derived mesenchymal stem cells (MSCs) and to investigate their biological characteristics. Methods Human umbilical cord blood (HUCB) was collected from full term deliveries scheduled for cesarean section, the cord blood mononuclear cells were isolated by lymphocyte separation medium, purified and cultured with Mesencult TM medium and acidic environment to produce adherent layer. Growth curves of MSCs were drawn and surface antigenic features were analyzed by flow cytometry technique. Results When HUCB-derived mononuclear cells were cultured in suitable medium, the adherent cells exhibited either an osteoclast- or mesenchymal-like phenotype cells produced with the MSCs,which displayed a fibroblast-like morphology and expressed several MSCs-related antigens CD29,CD44,CD105, but not CD34,CD45,CD106, HLA-DR. The cells were identical to human bone marrow-derived MSCs. Conclusion MSCs in HUCB can be cultured and expanded in vitro, and could be regarded as an alternative source of MSCs for experimental and clinical applications.