EN
Taking tissue engineering applications into clinical trials requires the development of effi cient, effective and safe protocols incorporated with effective 3-dimentional cell culturing and differentiation systems in order to develop transplantable tissues that may offer a life-line for many patients in the future. Umbilical cord blood, which is perhaps the most abundant world stem cell source, has shown previously practical and ethical advantages over other stem cells sources in many research and clinical applications including regenerative medicine. We developed a three step protocol for isolation, expansion and sequential neuronal differentiation and maturation of cord blood pluripotent stem cells (characterized with our unique triple immunocytochemisty scheme for Oct-4, Sox-2 and Nanog expression) in serum-free defi ned culturing conditions. We incorporated this protocol with 3-dimentional culturing system which produced properly organized neuronal tissues expressing Nestin and NF-200. We showed that umbilical cord blood pluripotent stem cells are a potential and promising candidate for future neural tissue engineering and regenerative medicine applications.