The invention provides a method for separating and culturing
umbilical cord blood mesenchymal stem cells. The method comprises the steps of carrying out secondary separation on
umbilical cord blood mononuclear cells by using a separating medium, and then inoculating the
umbilical cord blood mononuclear cells into a culture
bottle in which
fibronectin and CD90
monoclonal antibodies are coated; and culturing for 4-5 days by using a serum-free culture
system and simulating the in-vivo low-
oxygen growth environment of the mesenchymal stem cells, then, removing suspension cells, further culturing adherent cells, and subculturing after the
fusion rate of primary cells is up to 60%. By using the method for separating and culturing umbilical
cord blood mesenchymal stem cells, provided by the invention, the problems of adherence infirmness, low culture success rate, low purity and the like caused in the extraction and culture processes of the umbilical
cord blood mesenchymal stem cells are effectively solved, and the safety in clinical application is improved. In addition, the application ranges of the umbilical
cord blood mesenchymal stem cells are widened, the utilization values of the umbilical cord blood mesenchymal stem cells are developed, and the clinical application prospects of the umbilical cord blood mesenchymal stem cells are widened.