Ex Vivo Culture of Human Cranial Suture Cells

The culture of human cranial suture cells, including their osteoblasts, is an important asset to developmental and molecular biologists to allow study the molecular biology ex vivo. The use of cell cultures by bone biologists to investigate pathological bone formation has been well established (Marie et al. Vitro Cell Dev Biol 25:373-380, 1989), and the use of cell culture techniques was subsequently applied to investigate craniosynostosis (Marie. J Bone Miner Res 9(12):1847-1850, 1994). Cell cultures from fused, fusing, and fused cranial sutures allow comparative studies of cellular behavior from sutures with pathological craniosynostosis and those with unaffected sutures (Coussens et al. J Cell Physiol 218(1):183-191, 2009; Coussens et al. Differentiation 76(5):531-545, 2008).In addition to using this limited human resource for primary investigations, these human cell studies may be targeted to complement and help verify the findings of investigative studies undertaken using the more readily accessible animal cranial sutures. It is important, however, to remember that there may be critical differences in the animal genome which could impact on cellular function.This chapter describes the techniques for human suture cell culture and storage which have been used successfully since 2005 in the craniosynostosis laboratories in Adelaide.