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Neurosphere Formation, Migration, and Differentiation of Human Neural Stem Cells Cultured in Corning® Spheroid MicroplatesDownload
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September 21, 2015
Authors: Hilary Sherman, Hannah J. Gitschier, M.S., David H. Randle, Ph.D., Corning Incorporated, Life Sciences; Brad Larson, BioTek Instruments, Inc.
Neurosphere formation of neural stem cells (NSCs) is a widely used in vitro culture system and valuable model to study neurogenesis and neural development. This system allows for three dimensional expansion of NSCs within a more physiologically-relevant microenvironment. Having an easy to use, reproducible neurosphere culture and analysis method for studying NSC proliferation, migration, and neurotoxicity greatly enables their use for drug discovery and cell therapy applications. In this study, Corning® spheroid microplates were used for neurosphere formation, NSC proliferation, and migration in an easy-to-use format, amenable to high throughput screening. The microplates allow for NSC culture of uniform and directed neurosphere size over the course of 96 hours, throughout which multipotency is maintained, as assessed through Nestin and SOX2 marker expression.
Further, differentiation of NSCs into neuronal, astrocytic, and oligodendrocytic lineages was achieved via harvesting neurospheres and plating in high content imaging microplates with growth factors removed. Analysis of spheroid size, imaging of differentiated cells, and quantification of migration was accomplished with the BioTek® Cytation™ 5 Cell Imaging Multi- Mode Reader and Gen5™ Data Analysis Software.