Static and dynamic 3D culture of neural precursor cells on macroporous cryogel microcarriers
Ben Newland,
Fanny Ehret,
Franziska Hoppe,
Dimitri Eigel,
Dagmar Pette,
Heike Newland,
Petra B. Welzel,
Gerd Kempermann,
Carsten Werner
Affiliations
Ben Newland
Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials Dresden, 01069, Dresden, Germany; School of Pharmacy and Pharmaceutical Sciences, Cardiff University, CF10 3NB, Cardiff, UK; Corresponding author at: School of Pharmacy and Pharmaceutical Sciences, Cardiff University, CF10 3NB, Cardiff, UK
Fanny Ehret
German Center for Neurodegenerative Diseases (DZNE) Dresden, 01307, Dresden, Germany
Franziska Hoppe
Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials Dresden, 01069, Dresden, Germany
Dimitri Eigel
Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials Dresden, 01069, Dresden, Germany
Dagmar Pette
Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials Dresden, 01069, Dresden, Germany
Heike Newland
Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials Dresden, 01069, Dresden, Germany
Petra B. Welzel
Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials Dresden, 01069, Dresden, Germany
Gerd Kempermann
German Center for Neurodegenerative Diseases (DZNE) Dresden, 01307, Dresden, Germany; CRTD – Center for Regenerative Therapies Dresden, Technische Universität Dresden, 01307, Dresden, Germany
Carsten Werner
Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials Dresden, 01069, Dresden, Germany; CRTD – Center for Regenerative Therapies Dresden, Technische Universität Dresden, 01307, Dresden, Germany
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.
