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. 2021 Nov 27;10(12):3334. doi: 10.3390/cells10123334

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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General scheme of differentiation protocols used to produce NSC from PSC. (A) Example of an EB-based protocol. Undifferentiated PSC are grown in suspension in embryonic stem (ES) medium with ROCK inhibitor to form embryoid bodies (EB) that are further cultured in neural differentiation medium (NDM) containing N2 supplement, seeded on coated plates, and directed to form neural rosettes containing neuroepithelial cells. (B) Example of a dual SMAD inhibition-based protocol. Undifferentiated PSC are plated on Matrigel as single cells in an MEF (mouse embryonic fibroblasts)-conditioned medium (CM) and different inhibitors are added to the cells in the presence of knockout serum (KSR) medium. KSR medium is then gradually replaced by medium with N2 supplement. ROCK: Rho kinase; PLO-lam: poly-L-ornithine-laminin; RA: retinoic acid; FGF: fibroblast growth factor; EGF: epidermal growth factor; TGF-β: transforming growth factor beta.