Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 102, Issue 32, Pages 11331-11336Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0500010102
Keywords
regenerative medicine; differentiation; photoreceptor; induction; Six3
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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 Writ 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.
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