Culturing Neurons, Glia, and Progenitor Cells in Three-Dimensional Hydrogels

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


The physiologic environment of neuronal and glial cells is the three-dimensional (3D) tissue. We and others have demonstrated that more physiologically relevant cell responses occur in 3D culture environments as opposed to flat stiff culture substrates such as tissue culture plates and coverslips. Hydrogels provide significant advantages towards providing cells a biocompatible, all-encompassing culture environment, but pose challenges towards adapting standard biochemical and molecular biology assay techniques. Herein, we provide methods for encapsulating and culturing cells inside 3D type I collagen gels. We also provide several basic methods to assess cell viability, fix and stain cells while encapsulated in the gels, and digest the gels to retrieve live cells for subculturing or other experiments (e.g., flow cytometry).

Key words

Neurons Glia Progenitor cells Collagen gel Three-dimensional 


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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

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