Abstract
Protocols for culturing neural stem cells (NSCs) are increasingly finding utilization for studying and growing of tissues that can appropriately model the neural regeneration processes. Two-dimensional (2D) plastic or glass surface-enabled mammalian cell cultures have been the platforms for performing in vitro cell cultures. Isolated mammalian cells, however, come from three-dimensional (3D) spaces, thus recapitulating such 3D microenvironments is among the challenges for many tissue engineering applications. Herein, we present the protocols for culturing NSCs in 3D polysaccharide-based hydrogel microenvironments that mimic, for instance, the native extracellular matrix (ECM) space (of the brain). The protocols include three key steps: (1) generation of 3D hydrogels with living cells, (2) culturing NSCs in 3D environments, and (3) characterization via immunostaining and genetic expression assay (RT-qPCR).
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Acknowledgments
This work has been funded by Temple University and the GDFI Braincell Laboratory Co., Ltd., Seoul, South Korea.
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Jin, Gw., Ma, W., Suh, W.H. (2016). Three Dimensional Cell Culture of Human Neural Stem Cells Using Polysaccharide-Based Hydrogels and Subsequent Bioanalyses. In: Kye, M. (eds) MicroRNA Technologies. Neuromethods, vol 128. Humana Press, New York, NY. https://doi.org/10.1007/7657_2016_11
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DOI: https://doi.org/10.1007/7657_2016_11
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