Methods to study polyamine metabolism during osteogenesis

Mammalian polyamines, namely putrescine, spermidine, and spermine, have been implicated in many cellular homeostatic processes. Polyamines play a critical role in skeletal health as evidenced by recent studies and by skeletal disorders caused by polyamine imbalances, such as Snyder-Robinson Syndrome (SRS). However, very little is still known about the role of polyamines within bone development, homeostasis, and metabolism. Human bone marrow derived mesenchymal stromal cells (MSCs) provide a unique opportunity to study polyamines at a cellular and molecular level within the context of osteogenic differentiation and calcium deposition. Through in vitro work, mechanistic understanding of the role of polyamines within osteogenesis as well as the consequences of polyamine imbalance can provide new insights into potential therapeutics for those experiencing polyaminopathies. This chapter describes procedures to develop a human primary cell culture system and quantify osteoblastogenesis as a function of polyamine modulation.