Investigating Cell-ECM Interactions and ECM Synthesis in Three-Dimensional Hydrogels

  • Swarnalatha Balasubramanian
  • Elizabeth M. Powell
  • Jennie B. LeachEmail author
Part of the Neuromethods book series (NM, volume 93)


Model systems composed of cells cultured within three-dimensional (3D) gels are widely used to examine mechanisms of morphogenesis and repair and are applied as vehicles for delivering cells for therapeutic applications in vivo. Cells secrete extracellular matrix (ECM), the composition of which depends on the type of cell and cues present; the ECM is assembled and remodeled, forming a dynamic cellular niche. While it is generally recognized that the presence of ECM and cellular interactions with the ECM, for example via integrin binding, are closely tied to cell and tissue behavior, few investigations have centered on quantifying these processes in 3D gel microenvironments. Herein, we provide methods to characterize the location and patterns of ECM and cell-ECM interactions using immunofluorescence and collect cell lysate for analysis via immunoblotting or ELISA.

Key words

Neurons Glia ECM Integrins Collagen gel Three-dimensional 


  1. 1.
    Smith AM, Hunt NC, Shelton RM et al (2012) Alginate hydrogel has a negative impact on in vitro collagen 1 deposition by fibroblasts. Biomacromolecules 13:4032–4038PubMedGoogle Scholar
  2. 2.
    Mulyasasmita W, Lee JS, Heilshorn SC (2011) Molecular-level engineering of protein physical hydrogels for predictive sol-gel phase behavior. Biomacromolecules 12:3406–3411PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Legant WR, Chen CS, Vogel V (2012) Force-induced fibronectin assembly and matrix remodeling in a 3D microtissue model of tissue morphogenesis. Integr Biol (Camb) 4:1164–1174CrossRefGoogle Scholar
  4. 4.
    Ribeiro A, Balasubramanian S, Hughes D et al (2013) beta1-Integrin cytoskeletal signaling regulates sensory neuron response to matrix dimensionality. Neuroscience 248C:67–78CrossRefGoogle Scholar
  5. 5.
    Balasubramanian S, Powell EM, Leach JB (in preparation)Google Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Swarnalatha Balasubramanian
    • 1
  • Elizabeth M. Powell
    • 2
    • 3
    • 4
  • Jennie B. Leach
    • 1
    Email author
  1. 1.Department of Chemical, Biochemical & Environmental EngineeringUMBCBaltimoreUSA
  2. 2.Department of Anatomy & NeurobiologyUniversity of Maryland School of MedicineBaltimoreUSA
  3. 3.Department of PsychiatryUniversity of Maryland School of MedicineBaltimoreUSA
  4. 4.Department of BioengineeringUniversity of Maryland School of MedicineBaltimoreUSA

Personalised recommendations