Osteogenic differentiation of embryonic stem cells in 2D and 3D culture

Osteoblasts are the cells that contribute to the formation and function of bone tissue. Knowledge of their biology is important to understanding of the normal processes of bone repair, the development of diseases affecting bone tissue, and to the investigation of approaches to improve bone repair and to treat or prevent bone diseases. Osteoblasts can be readily isolated from bone tissues and grown in culture, and under relatively simple culture conditions, they will recapitulate many aspects of their normal biology. These culture conditions can be also applied to adult stem cells, such as mesenchymal/bone marrow stromal stem cells. More recently, these studies have been extended to include embryonic stem cells. This chapter provides detailed step-by-step protocols to investigate the differentiation of embryonic stem cells into osteoblasts. Several 2D and 3D culture methods are presented and enable comparisons to be made on the efficiency and mechanisms of osteogenic differentiation. Emphasis is also placed on methods to analyse and confirm osteogenic differentiation.