Substantial progress has been made in the development of methods to
direct embryonic stem cells to differentiate into various regions
of the central nervous system (CNS). We have used the current model
of eye specification to develop a protocol for directing human embryonic
stem cells to generate retinal progenitors and various types of retinal
neurons. Our method uses a multistep protocol in which embryoid bodies
are treated with inhibitors of both BMP signaling and canonical Wnt
signaling to promote expression of key eye field transcription factors
(EFTFs), as assayed by RT-PCR and immunofluorescence microscopy.
The retinal progenitor cells spontaneously undergo differentiation
into various types of retinal neurons, including photoreceptors,
and this can be promoted by treatment with small molecule inhibitors
of the Notch pathway.
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