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Generation of Rx(+)/Pax6(+) neural retinal precursors from embryonic stem cells

By Hanako Ikeda, Fumitaka Osakada, Kiichi Watanabe, Kenji Mizuseki, Tomoko Haraguchi, Hiroyuki Miyoshi, Daisuke Kamiya, Yoshihito Honda, Noriaki Sasai, Nagahisa Yoshimura, Masayo Takahashi and Yoshiki Sasai


We report directed differentiaion of retinal precursors in vitro from mouse ES cells. Six3(+) rostral brain progenitors are generated by culturing ES cells under serum-free suspension conditions (SFEB culture) in the presence of Wnt and Nodal antagonists (Dkk1 and LeftyA), and subsequently steered to differentiate into Rx(+) cells (16%) by treatment with activin and serum. Consistent with the characteristics of early neural retinal precursors, the induced Rx(+) cells coexpress Pax6 and the mitotic marker Ki67, but not Nestin. The ES cell-derived precursors efficiently generate cells with the photoreceptor phenotype (rhodopsin(+), recoverin(+)) when cocultured with embryonic retinal cells. Furthermore, organotypic culture studies demonstrate the selective integration and survival of ES cell-derived cells with the photoreceptor phenotype (marker expression and morphology) in the outer nuclear layer of the retina. Taken together, ES cells treated with SFEB/Dkk1/LeftyA/serum/activin generate neural retinal precursors, which have the competence of photoreceptor differentiation

Topics: Biological Sciences
Publisher: National Academy of Sciences
Year: 2005
DOI identifier: 10.1073/pnas.0500010102
OAI identifier:
Provided by: PubMed Central
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