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).
Neurons Glia Progenitor cells Collagen gel Three-dimensional
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