2024-03-28T16:39:28Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/1311012016-10-20T09:46:09Zcom_10261_134com_10261_1col_10261_1269
00925njm 22002777a 4500
dc
Erceg, Slaven
author
Lukovic, Dunja
author
Moreno-Manzano, Victoria
author
Stojkovic, Miodrag
author
Bhattacharya, Shom Shanker
author
2012
Here 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.
Current Protocols in Stem Cell Biology (Cap.1): Unit 1H.5 (2012)
http://hdl.handle.net/10261/131101
10.1002/9780470151808.sc01h05s20
Cerebellar granule neuron
Human pluripotent stem cell
Cerebellar neuron
Human embryonic stem cell
Derivation of cerebellar neurons from human pluripotent stem cells