Generation of Rx+/Pax6+ neural retinal precursors from embryonic stem cells.
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.
Ikeda
Hanako
H
Organogenesis and Neurogenesis Group, Center for Developmental Biology, RIKEN, Kobe 650-0047, Japan.
Osakada
Fumitaka
F
Watanabe
Kiichi
K
Mizuseki
Kenji
K
Haraguchi
Tomoko
T
Miyoshi
Hiroyuki
H
Kamiya
Daisuke
D
Honda
Yoshihito
Y
Sasai
Noriaki
N
Yoshimura
Nagahisa
N
Takahashi
Masayo
M
Sasai
Yoshiki
Y
D028341:Activins / Q000494:pharmacology; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D005136:Eye Proteins / Q000378:metabolism*; D005822:Genetic Vectors; D018398:Homeodomain Proteins / Q000378:metabolism*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D055458:Left-Right Determination Factors; D016086:Lentivirus; D051379:Mice; D009419:Nerve Tissue Proteins / Q000378:metabolism; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000378:metabolism*; D012097:Repressor Proteins / Q000378:metabolism*; D012160:Retina / Q000166:cytology* / Q000378:metabolism; D012243:Rhodopsin / Q000378:metabolism; D044967:Serum; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D000097607:Homeobox Protein SIX3
D028341:Activins; D016895:Culture Media, Serum-Free; D005136:Eye Proteins; D018398:Homeodomain Proteins; D036341:Intercellular Signaling Peptides and Proteins; D055458:Left-Right Determination Factors; D009419:Nerve Tissue Proteins; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; D012097:Repressor Proteins; D012243:Rhodopsin; D016212:Transforming Growth Factor beta; D000097607:Homeobox Protein SIX3; C501127:Dkk1 protein, mouse; C000605732:Pax6 protein, mouse; C501274:Rx protein, mouse