Intervertebral Disc Cell Mediated Mesenchymal Stem Cell Differentiation

¹ University of Manchester, Manchester, United Kingdom
² University of Liverpool, Liverpool, United Kingdom

^* To whom correspondence should be addressed. E-mail: judith.hoyland{at}manchester.ac.uk.

	Abstract

Low back pain is one of the largest health problems in the western world today and intervertebral disc degeneration has been identified as a main cause. Currently treatments are symptomatic but cell-based tissue engineering methods are realistic alternatives for tissue regeneration. However the major problem for these strategies is the generation of a suitable population of cells. Adult bone marrow-derived mesenchymal stem cells are undifferentiated, multipotent cells that have the ability to differentiate into a number of cell types, including the chondrocyte-like cells found within the nucleus pulposus of the intervertebral disc. However, no method exists to differentiate these cells in an accessible monolayer environment. We have conducted co-culture experiments to determine whether cells from the human nucleus pulposus can initiate the differentiation of human mesenchymal stem cells either with or without cell-cell contact. Fluorescent labelling of the stem cell population and high-speed cell sorting following co-culture with cell-cell contact allowed examination of individual cell populations. Real-time quantitative PCR showed significant increases in nucleus pulposus-marker genes in stem cells when cells were co-cultured with contact for 7 days which was modulated by cell ratio. No significant change in nucleus pulposus-marker gene expression in either nucleus pulposus cells or stem cells was observed when cells were cultured without contact, regardless of cell ratio. Thus we have shown that human nucleus pulposus and mesenchymal stem cell co-culture with contact is a viable method for generating a large population of differentiated cells that could be used in cell-based tissue engineering therapies for regeneration of the degenerate intervertebral disc.

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