We use cookies to improve your experience. By continuing to browse this site, you accept our cookie policy.×
Skip main navigation
Aging Health
Bioelectronics in Medicine
Biomarkers in Medicine
Breast Cancer Management
CNS Oncology
Colorectal Cancer
Concussion
Epigenomics
Future Cardiology
Future Medicine AI
Future Microbiology
Future Neurology
Future Oncology
Future Rare Diseases
Future Virology
Hepatic Oncology
HIV Therapy
Immunotherapy
International Journal of Endocrine Oncology
International Journal of Hematologic Oncology
Journal of 3D Printing in Medicine
Lung Cancer Management
Melanoma Management
Nanomedicine
Neurodegenerative Disease Management
Pain Management
Pediatric Health
Personalized Medicine
Pharmacogenomics
Regenerative Medicine
Research ArticleFree Access

Co-culture induces mesenchymal stem cell differentiation and modulation of the degenerate human nucleus pulposus cell phenotype

    Sandra Strassburg

    Regenerative Medicine, School of Biomedicine, Faculty of Medical & Human Sciences, The University of Manchester, Oxford Road, Manchester, M13 9PT, UK

    ,
    Stephen M Richardson

    Regenerative Medicine, School of Biomedicine, Faculty of Medical & Human Sciences, The University of Manchester, Oxford Road, Manchester, M13 9PT, UK

    ,
    Anthony J Freemont

    Regenerative Medicine, School of Biomedicine, Faculty of Medical & Human Sciences, The University of Manchester, Oxford Road, Manchester, M13 9PT, UK

    &
    Published Online:https://doi.org/10.2217/rme.10.59

    Aims: While mesenchymal stem cell (MSC)-based therapies for repair of the degenerate intervertebral disc (IVD) have been proposed, the interaction of MSCs with cells of the degenerate IVD has not been fully investigated. Therefore, it is unclear whether implanted MSCs would differentiate into nucleus pulposus (NP) cells and/or stimulate endogenous NP cells. Here, we investigate the differences in interaction between human MSCs and NP cells from both nondegenerate and degenerate discs during in vitro co-culture with direct cell–cell contact. Materials & methods : Human bone marrow-derived MSCs (labeled with CFDA) were co-cultured with direct cell–cell contact in monolayer with NP cells obtained from nondegenerate or degenerate human NP tissue from lumbar IVDs at 50:50 ratios for 7 days. Differentiation of MSCs and changes of matrix-associated genes in NP cells were assessed by quantitative real-time PCR. Results: MSCs differentiated to an NP-like phenotype following direct co-culture with both nondegenerate and degenerate NP, as shown by a significant upregulation of SOX9, type VI collagen, aggrecan and versican gene expression together with a simultaneous upregulation of CDMP-1, TGF-β1, IGF-1 and CTGF. Direct co-culture of normal NP cells with MSCs had no effect on the phenotype of normal NP cells, while co-culture with degenerate NP cells resulted in enhanced matrix gene expression in degenerate NP cells, accompanied by increases in both TGF-β and CDMP-1 gene expression. Conclusion: Importantly for MSC-based therapies for repair of the degenerate IVD, these data suggest that cellular interactions between MSCs and degenerate NP cells may both stimulate MSC differentiation to an NP-like phenotype and also stimulate the endogenous NP cell population to regain a nondegenerate phenotype and consequently enhance matrix synthesis for self-repair.

    Bibliography

    • Waddell G: Low back pain: a twentieth century health care enigma. Spine21(24),2820–2825 (1996).
    • Luoma K, Riihimaki H, Luukkonen R, Raininko R, Viikari-Juntura E, Lamminen A: Low back pain in relation to lumbar disc degeneration. Spine25(4),487–492 (2000).
    • Cheung KM, Karppinen J, Chan D et al.: Prevalence and pattern of lumbar magnetic resonance imaging changes in a population study of one thousand forty-three individuals. Spine34(9),934–940 (2009).
    • Sive JI, Baird P, Jeziorsk M, Watkins A, Hoyland JA, Freemont AJ: Expression of chondrocyte markers by cells of normal and degenerate intervertebral discs. Mol. Pathol.55(2),91–97 (2002).
    • Melrose J, Ghosh P, Taylor TK: A comparative analysis of the differential spatial and temporal distributions of the large (aggrecan, versican) and small (decorin, biglycan, fibromodulin) proteoglycans of the intervertebral disc. J. Anat.198(Pt 1),3–15 (2001).
    • Roberts S, Ayad S, Menage PJ: Immunolocalisation of type VI collagen in the intervertebral disc. Ann. Rheum. Dis.50(11),787–791 (1991).
    • Urban MR, Fairbank JC, Bibby SR, Urban JP: Intervertebral disc composition in neuromuscular scoliosis: changes in cell density and glycosaminoglycan concentration at the curve apex. Spine (Phila Pa 1976), 26(6),610–617 (2001).
    • Le Maitre CL, Pockert A, Buttle DJ, Freemont AJ, Hoyland JA: Matrix synthesis and degradation in human intervertebral disc degeneration. Biochem. Soc. Trans.35(Pt 4),652–655 (2007).
    • Pockert AJ, Richardson SM, Le Maitre CL et al.: Modified expression of the ADAMTS enzymes and tissue inhibitor of metalloproteinases 3 during human intervertebral disc degeneration. Arthritis Rheum.60(2),482–491 (2009).
    • 10  Freemont AJ, Watkins A, Le Maitre C, Jeziorska M, Hoyland JA: Current understanding of cellular and molecular events in intervertebral disc degeneration: implications for therapy. J. Pathol.196(4),374–379 (2002).
    • 11  Thompson JP, Oegema TR Jr, Bradford DS: Stimulation of mature canine intervertebral disc by growth factors. Spine16(3),253–260 (1991).
    • 12  Chujo T, An HS, Akeda K et al.: Effects of growth differentiation factor-5 on the intervertebral disc – in vitro bovine study and in vivo rabbit disc degeneration model study. Spine31(25),2909–2917 (2006).
    • 13  Le Maitre CL, Freemont AJ, Hoyland JA: The role of interleukin-1 in the pathogenesis of human intervertebral disc degeneration. Arthritis Res. Ther.7(4),R732–R745 (2005).
    • 14  Okuma M, Mochida J, Nishimura K, Sakabe K, Seiki K: Reinsertion of stimulated nucleus pulposus cells retards intervertebral disc degeneration: an in vitro and in vivo experimental study. J. Orthop. Res.18(6),988–997 (2000).
    • 15  Gruber HE, Johnson TL, Leslie K et al.: Autologous intervertebral disc cell implantation: a model using Psammomys obesus, the sand rat. Spine27(15),1626–1633 (2002).
    • 16  Nomura T, Mochida J, Okuma M, Nishimura K, Sakabe K: Nucleus pulposus allograft retards intervertebral disc degeneration. Clin. Orthop. Relat. Res.389,94–101 (2001).
    • 17  Le Maitre CL, Freemont AJ, Hoyland JA: Accelerated cellular senescence in degenerate intervertebral discs: a possible role in the pathogenesis of intervertebral disc degeneration. Arthritis Res. Ther.9(3),R45 (2007).
    • 18  Roberts S, Evans EH, Kletsas D, Jaffray DC, Eisenstein SM: Senescence in human intervertebral discs. Eur. Spine J.15(Suppl. 3),S312–S316 (2006).
    • 19  Le Maitre CL, Freemont AJ, Hoyland JA: Localization of degradative enzymes and their inhibitors in the degenerate human intervertebral disc. J. Pathol.204(1),47–54 (2004).
    • 20  Pittenger MF, Mackay AM, Beck SC et al.: Multilineage potential of adult human mesenchymal stem cells. Science284(5411),143–147 (1999).
    • 21  Richardson SM, Hughes N, Hunt JA, Freemont AJ, Hoyland JA: Human mesenchymal stem cell differentiation to NP-like cells in chitosan–glycerophosphate hydrogels. Biomaterials29(1),85–93 (2008).
    • 22  Risbud MV, Albert TJ, Guttapalli A et al.: Differentiation of mesenchymal stem cells towards a nucleus pulposus-like phenotype in vitro: implications for cell-based transplantation therapy. Spine29(23),2627–2632 (2004).
    • 23  Crevensten G, Walsh AJ, Ananthakrishnan D et al.: Intervertebral disc cell therapy for regeneration: mesenchymal stem cell implantation in rat intervertebral discs. Ann. Biomed. Eng.32(3),430–434 (2004).
    • 24  Sakai D, Mochida J, Yamamoto Y et al.: Transplantation of mesenchymal stem cells embedded in Atelocollagen gel to the intervertebral disc: a potential therapeutic model for disc degeneration. Biomaterials24(20),3531–3541 (2003).
    • 25  Richardson SM, Walker RV, Parker S et al.: Intervertebral disc cell-mediated mesenchymal stem cell differentiation. Stem Cells24(3),707–716 (2006).
    • 26  Yamamoto Y, Mochida J, Sakai D et al.: Upregulation of the viability of nucleus pulposus cells by bone marrow-derived stromal cells: significance of direct cell-to-cell contact in coculture system. Spine29(14),1508–1514 (2004).
    • 27  Vadala G, Studer RK, Sowa G et al.: Coculture of bone marrow mesenchymal stem cells and nucleus pulposus cells modulate gene expression profile without cell fusion. Spine33(8),870–876 (2008).
    • 28  Sobajima S, Vadala G, Shimer A, Kim JS, Gilbertson LG, Kang JD: Feasibility of a stem cell therapy for intervertebral disc degeneration. Spine J.8(6),888–896 (2008).
    • 29  Yang SH, Wu CC, Shih TT, Sun YH, Lin FH: In vitro study on interaction between human nucleus pulposus cells and mesenchymal stem cells through paracrine stimulation. Spine33(18),1951–1957 (2008).
    • 30  Svanvik T, Barreto Henriksson H, Karlsson C, Hagman M, Lindahl A, Brisby H: Human disk cells from degenerated disks and mesenchymal stem cells in co-culture result in increased matrix production. Cells Tissues Organs191(1),2–11 (2010).
    • 31  Lu ZF, Zandieh Doulabi B, Wuisman PI, Bank RA, Helder MN: Differentiation of adipose stem cells by nucleus pulposus cells: configuration effect. Biochem. Biophys. Res. Commun.359(4),991–996 (2007).
    • 32  Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods25(4),402–408 (2001).
    • 33  Henriksson HB, Svanvik T, Jonsson M et al.: Transplantation of human mesenchymal stems cells into intervertebral discs in a xenogeneic porcine model. Spine (Phila Pa 1976)34(2),141–148 (2009).
    • 34  Caplan AI, Dennis JE: Mesenchymal stem cells as trophic mediators. J. Cell Biochem.98(5),1076–1084 (2006).
    • 35  Feng G, Wan Y, Balian G, Laurencin CT, Li X: Adenovirus-mediated expression of growth and differentiation factor-5 promotes chondrogenesis of adipose stem cells. Growth Factors26(3),132–142 (2008).
    • 36  Kawamura K, Chu CR, Sobajima S et al.: Adenoviral-mediated transfer of TGF-β1 but not IGF-1 induces chondrogenic differentiation of human mesenchymal stem cells in pellet cultures. Exp. Hematol.33(8),865–872 (2005).
    • 37  Bai X, Xiao Z, Pan Y et al.: Cartilage-derived morphogenetic protein-1 promotes the differentiation of mesenchymal stem cells into chondrocytes. Biochem. Biophys. Res. Commun.325(2),453–460 (2004).
    • 38  Steinert AF, Palmer GD, Pilapil C, Ulrich N, Evans CH, Ghivizzani SC: Enhanced in vitro chondrogenesis of primary mesenchymal stem cells by combined gene transfer. Tissue Eng. Part A15(5),1127–1139 (2009).
    • 39  Indrawattana N, Chen G, Tadokoro M et al.: Growth factor combination for chondrogenic induction from human mesenchymal stem cell. Biochem. Biophys. Res. Commun.320(3),914–919 (2004).
    • 40  Djouad F, Delorme B, Maurice M et al.: Microenvironmental changes during differentiation of mesenchymal stem cells towards chondrocytes. Arthritis Res. Ther.9(2),R33 (2007).
    • 41  Li X, Leo BM, Beck G, Balian G, Anderson GD: Collagen and proteoglycan abnormalities in the GDF-5-deficient mice and molecular changes when treating disk cells with recombinant growth factor. Spine29(20),2229–2234 (2004).
    • 42  Sakai D, Mochida J, Iwashina T et al.: Regenerative effects of transplanting mesenchymal stem cells embedded in atelocollagen to the degenerated intervertebral disc. Biomaterials27(3),335–345 (2006).
    • 43  Zhang YG, Guo X, Xu P, Kang LL, Li J: Bone mesenchymal stem cells transplanted into rabbit intervertebral discs can increase proteoglycans. Clin. Orthop. Relat. Res.430,219–226 (2005).