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Journal for Biophysical Chemistry

Co-culture of osteocytes and neurons on a unique patterned surface

Abstract

Neural and skeletal communication is essential for the maintenance of bone mass and transmission of pain, yet the mechanism(s) of signal transduction between these tissues is unknown. The authors established a novel system to co-culture murine long bone osteocyte-like cells (MLO-Y4) and primary murine dorsal root ganglia (DRG) neurons. Assessment of morphology and maturation marker expression on perlecan domain IV peptide (PlnDIV) and collagen type-1 (Col1) demonstrated that PlnDIV was an optimal matrix for MLO-Y4 culture. A novel matrix-specificity competition assay was developed to expose these cells to several extracellular matrix proteins such as PlnDIV, Col1, and laminin (Ln). The competition assay showed that approximately 70% of MLOY4 cells preferred either PlnDIV or Col1 to Ln. To co-culture MLO-Y4 and DRG, we developed patterned surfaces using micro-contact printing to create 40 μm × 1 cm alternating stripes of PlnDIV and Ln or PlnDIV and Col1. Co-culture on PlnDIV/Ln surfaces demonstrated that these matrix molecules provided unique cues for each cell type, with MLO-Y4 preferentially attaching to the PlnDIV lanes and DRG neurons to the Ln lanes. Approximately 80% of DRG were localized to Ln. Cellular processes from MLO-Y4 were closely associated with axonal extensions of DRG neurons. Approximately 57% of neuronal processes were in close proximity to nearby MLO-Y4 cells at the PlnDIV-Ln interface. The surfaces in this new assay provided a unique model system with which to study the communication between osteocyte-like cells and neurons in an in vitro environment.

References

  1. C. Adam, A. Llorens, B. Baroukh, M. Cherruau, and J. L. Saffar, Exp. Physiol. 85, 61 (2000).

    Article  CAS  Google Scholar 

  2. H. M. Blau and D. Baltimore, J. Cell Biol. 112, 781 (1991).

    Article  CAS  Google Scholar 

  3. D. E. Ingber, J. Cell Sci. 104, 613 (1993).

    Google Scholar 

  4. S. M. Frisch and H. Francis, J. Cell Biol. 124, 619 (1994).

    Article  CAS  Google Scholar 

  5. E. H. Sage, J. Clin. Invest. 107, 781 (2001).

    Article  CAS  Google Scholar 

  6. W. R. Thompson, S. Modla, B. J. Grindel, K. J. Czymmek, C. B. Kirn-Safran, L. Wang, R. L. Duncan, and M. C. Farach-Carson, J. Bone Miner. Res. 26, 618 (2011).

    Article  CAS  Google Scholar 

  7. E. Arikava-Hirasawa, H. Watanabe, H. Takami, J. R. Hassell, and Y. Yamada, Nat. Genet. 23, 354 (1999).

    Article  Google Scholar 

  8. H. Y. Zhou, Ann. N.Y. Acad. Sci. 1116, 323 (2007).

    Article  CAS  Google Scholar 

  9. K. Q. Zhang, C. Barragan-Adjemian, L. Ye, S. Kotha, M. Dallas, Y. B. Lu, S. J. Zhao, M. Harris, S. E. Harris, J. Q. Feng, and L. F. Bonewald, Mol. Cell. Biol. 26, 4539 (2006).

    Article  CAS  Google Scholar 

  10. T. A. Franz-Odendaal, B. K. Hall, and P. E. Witten, Dev. Dyn. 235, 176 (2006).

    Article  CAS  Google Scholar 

  11. A. Martin, V. David, J. S. Laurence, P. M. Schwarz, E. M. Lafer, A. M. Hedge, and P. S. N. Rowe, Endocrinology 149, 1757 (2008).

    Article  CAS  Google Scholar 

  12. H. F. Rios, L. Ye, V. Dusevich, D. Eick, L. F. Bonewald, and J. Q. Feng, J. Musculoskeletal and Neuronal Interact. 5, 325 (2005).

    CAS  Google Scholar 

  13. J. Q. Feng, R. D'Souza, and C. Qin, J. Dent. Res. 86, 1134 (2007).

    Article  Google Scholar 

  14. C. Y. Lu, S. Huang, T. Miclau, J. A. Helms, and C. Colnot, Histochem. Cell Biol. 121, 493 (2004).

    Article  CAS  Google Scholar 

  15. A. Nampei, J. Hashimoto, K. Hayashida, H. Tsuboi, K. Shi, H. Miyashita, T. Yamada, S. Morimoto, T. Ochi, and H. Yoshikawa, Bone 32, S136 (2003).

    Article  Google Scholar 

  16. S. Y. Fu and T. Gordon, Mol. Neurobiol. 14, 67 (1997).

    Article  CAS  Google Scholar 

  17. P. C. Letourneau, M. L. Condic, and D. M. Snow, Curr. Opin. Genet. Dev. 2, 625 (1992).

    Article  CAS  Google Scholar 

  18. P. Kannus, L. Jozsa, T. A. H. Jarvinen, T. L. N. Jarvinen, M. Kvist, A. Natri, and M. Jarvinen, Histochem. J. 30, 799 (1998).

    Article  CAS  Google Scholar 

  19. D. Mach, S. Rogers, M. Sabino, N. Luger, M. Schwel, J. Pomonis, C. Keyser, D. Clohisy, D. Adams, P. O'Leary, and P. Manyth, Neuroscience 113, 155 (2002).

    Article  CAS  Google Scholar 

  20. S. Vukicevic, F. P. Luyten, H. K. Kleinman, and A. H. Reddi, Cell 63, 437 (1990).

    Article  CAS  Google Scholar 

  21. C. M. Serre, D. Farlay, P. D. Delmas, and C. Chenu, Bone 25, 623 (1999).

    Article  CAS  Google Scholar 

  22. C. A. Goubko and X. D. Cao, Mater. Sci. Eng., C 29, 1855 (2009).

    Article  CAS  Google Scholar 

  23. M. Yamato, O. H. Kwon, M. Hirose, A. Kikuchi, and T. Okano, J. Biomed. Mater. Res. 55, 137 (2001).

    Article  CAS  Google Scholar 

  24. J. Y. Lee, S. S. Shah, J. Yan, M. C. Howland, A. N. Parikh, T. R. Pan, and A. Revzin, Langmuir 25, 3880 (2009).

    Article  CAS  Google Scholar 

  25. Z. P. Zhang, R. Yoo, M. Wells, T. P. Beebe, R. Biran, and P. Tresco, Biomaterials 26, 47 (2005).

    Article  Google Scholar 

  26. M. C. Farach-Carson, A. J. Brown, M. Lynam, J. B. Safran, and D. D. Carson, Matrix Biol. 27, 150 (2008).

    Article  CAS  Google Scholar 

  27. C. H. Turner, A. G. Robling, R. L. Duncan, and D. B. Burr, Calcif. Tissue Int. 70, 435 (2002).

    Article  CAS  Google Scholar 

  28. M. S. Patel and F. Elefteriou, Calcif. Tissue Int. 81, 247 (2007).

    Article  CAS  Google Scholar 

  29. W. M. Theilacker, H. Bui, and T. P. Beebe, BioInterphases 6, 105 (2011).

    Article  CAS  Google Scholar 

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Correspondence to Thomas P. Beebe Jr..

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Boggs, M.E., Thompson, W.R., Farach-Carson, M.C. et al. Co-culture of osteocytes and neurons on a unique patterned surface. Biointerphases 6, 200–209 (2011). https://doi.org/10.1116/1.3664050

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  • DOI: https://doi.org/10.1116/1.3664050