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Derivation of cerebellar neurons from human pluripotent stem cells

AutorErceg, Slaven; Lukovic, Dunja ; Moreno-Manzano, Victoria; Stojkovic, Miodrag; Bhattacharya, Shom Shanker
Palabras claveCerebellar granule neuron
Human pluripotent stem cell
Cerebellar neuron
Human embryonic stem cell
Fecha de publicación2012
EditorJohn Wiley & Sons
CitaciónCurrent Protocols in Stem Cell Biology (Cap.1): Unit 1H.5 (2012)
ResumenHere we provide a protocol for differentiation of human embryonic stem cells (hESC) into cerebellar neurons using a novel defined culture method. This protocol is based on the application of inductive signaling factors involved in the early patterning of the cerebellar region of the neural tube, followed by the application of factors responsible for cerebellar neuron specification. Human pluripotent stem cells are induced to form spherical embryonic-like structures called embryoid bodies (EBs) and neuroepithelial tube-like rosettes using defined chemical conditions. In the presence of FGF, Wnt, and RA signaling factors the rosettes were specified to OTX2-expressing cells. Further specification of derived cells involves application of BMP factors involved in early development of granule cell progenitors, followed by mitogens and neurotrophins. It typically takes 5 weeks to generate the functional cerebellar granule neurons. This protocol is feeder-free, applies human recombinant factors, and produces high yield of desired neurons.
URIhttp://hdl.handle.net/10261/131101
DOI10.1002/9780470151808.sc01h05s20
Identificadoresdoi: 10.1002/9780470151808.sc01h05s20
isbn: 9780470151808
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