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.

STAR Protocols, 2021


This protocol describes the derivation of neuroepithelial stem (NES) cells from human induced pluripotent stem cells. NES cells can be further differentiated into neurons and glia. The PSC culture and NES differentiation were done on plates coated with Biolaminin 521 (laminin-521). To avoid clonal selection of isolated NES cells, it is recommended to follow the culture conditions described. The protocol is expected to result in highly proliferate NES cells providing a good source of cells of a neuronal cell lineage. Glial cells are formed after approximately 45 days of differentiation.