Isolation of mesenchymal stem cells from human cord blood

Cord blood is a rich source of stem cells especially for hematopoietic stem cells. Recently, mesenchymal stem cells (MSCs) have also been shown to exist in cord blood. However, these fibroblast-like multipotent progenitor cells are rather rare in cord blood. Many different methods have been used for their culture. This unit describes one method to obtain MSCs from cord blood and another method to differentiate these cells into osteoblasts, which is one of the lineages that mesenchymal stem cells are capable of differentiating into. The starting material for the protocol is cord blood-derived mononuclear cells. As cord blood contains a great number of erythroid precursors, the glycophorin A-positive cells are depleted using magnetic cell separation to reduce their presence in MSC culture. Osteoblast differentiation and a method to demonstrate the result of the differentiation are also described in this unit.