Abstract
Neural precursor cells have been much studied to further our understanding of the far-reaching and controversial question of adult neurogenesis. Currently, differentiation of primary neural precursor cells from the mouse dentate gyrus via 2-dimentional in vitro culture yields low numbers of neurons, a major hindrance to the field of study. 3-dimentional “neurosphere” culture allows better 3D cell-cell contact, but control over cell differentiation is poor because nutrition and oxygen restrictions at the core of the sphere causes spontaneous differentiation, predominantly to glial cells, not neurons. Our group has developed macroporous scaffolds, which overcome the above-mentioned problems, allowing long-term culture of neural stem cells, which can be differentiated into a much higher yield of neurons. Herein we describe a method for culturing neural precursor cells on RGD peptide functionalized-heparin containing cryogel scaffolds, either in standard non-adherent well-plates (static culture) or in spinner flasks (dynamic culture). This method includes:•The synthesis and characterization of heparin based microcarriers.•A “static” 3D culture method for that does not require spinner flask equipment.•“Dynamic” culture in which cell loaded microcarriers are transferred to a spinner flask.
Highlights
Method ArticleStatic and dynamic 3D culture of neural precursor cells on macroporous cryogel microcarriers
Neural precursor cells have been much studied to further our understanding of the far-reaching and controversial question of adult neurogenesis
The cryogel microcarriers were prepared in a manner similar to that reported previously [1]
Summary
Static and dynamic 3D culture of neural precursor cells on macroporous cryogel microcarriers. Ben Newlanda,b,*, Fanny Ehretc, Franziska Hoppea, Dimitri Eigela, Dagmar Pettea, Heike Newlanda, Petra B. Gerd Kempermannc,d, Carsten Wernera,d a Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials Dresden, 01069, Dresden, Germany b School of Pharmacy and Pharmaceutical Sciences, Cardiff University, CF10 3NB, Cardiff, UK c German Center for Neurodegenerative Diseases (DZNE) Dresden, 01307, Dresden, Germany d CRTD – Center for Regenerative Therapies Dresden, Technische Universität Dresden, 01307, Dresden, Germany
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