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标题：Static and dynamic 3D culture of neural precursor cells on macroporous cryogel microcarriers
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作者：Ben Newland ; Fanny Ehret ; Franziska Hoppe 等
期刊名称：MethodsX
印刷版ISSN：2215-0161
电子版ISSN：2215-0161
出版年度：2020
卷号：7
页码：1-5
DOI：10.1016/j.mex.2020.100805
语种：English
出版社：Elsevier
摘要：Graphical abstractDisplay OmittedAbstractNeural 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-dimentionalin vitroculture 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.
关键词：Scaffolds;Neural progenitor differentiation;Cell survival;Dentate gyrus;Heparin;Biomaterials for cell culture