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TISSUE-SPECIFIC STEM CELLS

Intervertebral Disc Cell–Mediated Mesenchymal Stem Cell Differentiation

^a UK Centre for Tissue Engineering, Division of Laboratory and Regenerative Medicine, University of Manchester, Manchester, United Kingdom;
^b UK Centre for Tissue Engineering, Clinical Engineering, University of Liverpool, Liverpool, United Kingdom

Key Words. Mesenchymal stem cell • Nucleus pulposus • Chondrocyte-like • Coculture • Differentiation

Correspondence: Judith A. Hoyland, Ph.D., Division of Laboratory and Regenerative Medicine, Stopford Building, University of Manchester, Oxford Road, Manchester, M13 9PT, U.K. Telephone: +44(0)161-275-5425; Fax: +44(0)161-275-5289; e-mail: judith.hoyland{at}manchester.ac.uk

Received May 5, 2005; accepted for publication October 5, 2005.
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 (MSCs) 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 (NP) of the intervertebral disc; however, no method exists to differentiate these cells in an accessible monolayer environment. We have conducted coculture experiments to determine whether cells from the human NP can initiate the differentiation of human MSCs with or without cell–cell contact. Fluorescent labeling of the stem cell population and high-speed cell sorting after coculture with cell–cell contact allowed examination of individual cell populations. Real-time quantitative polymerase chain reaction showed significant increases in NP marker genes in stem cells when cells were cocultured with contact for 7 days, and this change was regulated by cell ratio. No significant change in NP marker gene expression in either NP cells or stem cells was observed when cells were cultured without contact, regardless of cell ratio. Thus, we have shown that human NP and MSC coculture 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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