Intervertebral Disc Whole Organ Cultures
This chapter aims to provide an overview of the main morphological changes observed in human degenerated intervertebral disc (IVD), the established ex vivo organ culture models aiming to simulate these changes, and the methods used to monitor these simulated changes. It focuses on the methodology of measured output parameters in degenerative organ cultures with the aim to provide guidance on the choice of the most appropriate model for future experiments. Different cell and organ culture methods are used to mimic and investigate the mechanisms involved in human IVD degeneration. Besides the synthesis rate or the total amount of deposed glycosaminoglycan, the phenotype of the IVD cells can be assessed using gene expression. The most popular method to determine the total collagen content of an IVD tissue sample is the hydroxyproline assay. It is a quantitative measurement of the amino acid hydroxyproline, representing a major component of all triple-helical collagen proteins and functioning as an enhancer of collagen stability.