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.
School of Pharmacy, Centre for Biomolecular Science, University of Nottingham, Nottingham, UK.
Lee
L
Buttery
Robert
R
Bielby
Daniel
D
Howard
Kevin
K
Shakesheff
D000464:Alginates / Q000494:pharmacology; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001681:Biological Assay; D015820:Cadherins / Q000378:metabolism; D002449:Cell Aggregation / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D058732:Embryoid Bodies / Q000166:cytology / Q000187:drug effects; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D005786:Gene Expression Regulation / Q000187:drug effects; D020723:Glucuronic Acid / Q000494:pharmacology; D006603:Hexuronic Acids / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008856:Microscopy, Fluorescence; D008903:Minerals / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects; D011091:Polyesters / Q000494:pharmacology; D013194:Staining and Labeling; D054457:Tissue Scaffolds / Q000737:chemistry; D055114:X-Ray Microtomography
D000464:Alginates; D015820:Cadherins; D006603:Hexuronic Acids; D008903:Minerals; D011091:Polyesters; C033616:poly(lactide); D020723:Glucuronic Acid; D000469:Alkaline Phosphatase