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

2005; National Academy of Sciences; Volume: 102; Issue: 32 Linguagem: Inglês

10.1073/pnas.0500010102

1091-6490

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

Pluripotent Stem Cells Research

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.