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Protocol for the derivation, culturing, and differentiation of human iPS-cell-derived neuroepithelial stem cells to study neural differentiation in vitro

Javier Calvo‐Garrido, Dania Winn, Camilla Maffezzini, Anna Wedell, Christoph Freyer, Anna Falk, Anna Wredenberg

2021STAR Protocols56 citationsDOIOpen Access PDF

Abstract

Here, we present a revised protocol to derive neuroepithelial stem (NES) cells from human induced pluripotent stem cells. NES cells can be further differentiated into a culture of neurons (90%) and glia (10%). We describe how to derive and maintain NES cells in culture and how to differentiate them. In addition, we show the potential use of NES cells to study the role of reactive oxygen species in neuronal differentiation and a guideline for NES cell transfection. For complete details on the use and execution of this protocol, please refer to Calvo-Garrido et al. (2019); Falk et al. (2012).

Topics & Concepts

Neuroepithelial cellInduced pluripotent stem cellNeural stem cellCellular differentiationStem cellCell biologyBiologyIn vitroHuman Induced Pluripotent Stem CellsCell cultureNeuroscienceEmbryonic stem cellGeneticsGenePluripotent Stem Cells ResearchNeuroscience and Neural Engineering3D Printing in Biomedical Research