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

Academic Article

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 MLO-Y4 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.
  • Keywords

  • Animals, Biomarkers, Cell Shape, Cells, Cultured, Coculture Techniques, Collagen Type I, Cross-Linking Reagents, Ganglia, Spinal, Gene Expression Regulation, Heparan Sulfate Proteoglycans, Laminin, Mice, Microscopy, Confocal, Neurons, Osteocytes, Peptides, Photoelectron Spectroscopy, Protein Structure, Tertiary, RNA, Messenger, Surface Properties
  • Digital Object Identifier (doi)

    Author List

  • Boggs ME; Thompson WR; Farach-Carson MC; Duncan RL; Beebe TP
  • Start Page

  • 200
  • End Page

  • 209
  • Volume

  • 6
  • Issue

  • 4