Year: 2011
Type: 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.
Source: Biointerphases
DOI: 10.1116/1.3664050
Authors: Mary E. Boggs, William R. Thompson, Mary C. Farach-Carson, Randall L. Duncan, Thomas P. Beebe
Institutions: University of Delaware, Rice University
Language: English
Subfield: Cell Biology
Domain: Life Sciences
Open Access status: hybrid
Funders: National Science Foundation, National Institutes of Health, U.S. Public Health Service, National Cancer Institute, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institute of Biomedical Imaging and Bioengineering, Division of Materials Research, National Center for Research Resources, Division of Chemistry, Eunice Kennedy Shriver National Institute of Child Health and Human Development
Awards: COBRE for Women in Science & Engineering on Osteoarthritis, G1822187544, G2767958271, Mechanotransduction: Puringergic Signaling in Bone, G6326362866, G655362773, GOALI: Solution-based Fluoropolymer Films, G7547665281, Acquisition of a Time-of-Flight Secondary-Ion Mass Spectrometer Surface Analysis System, Probing Single-Molecule Neuron-Ligand Pathfinding, PT/PHD Predoctoral Training Program
OpenAlex ID: W2032241908