Generation of Rx+/Pax6+ neural retinal precursors from embryonic stem cells
Generation of Rx+/Pax6+ neural retinal precursors from embryonic stem cells
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DOI：
10.1073/pnas.0500010102
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发表时间：
2005-08-09
影响因子：
11.1
通讯作者：
Sasai, Y
中科院分区：
文献类型：
--
作者：
Ikeda, H;Osakada, F;Sasai, Y
我们报道了体外培养的视网膜前体细胞与小鼠ES细胞的定向分化。在无血清悬浮条件下(SFEB培养)，在WRIT和Nodal拮抗剂(Dkk1和LeftyA)的存在下培养ES细胞，生成Six3(+)吻侧脑祖细胞，然后通过激活素和血清诱导分化为Rx(+)细胞(16%)。与早期视网膜神经前体细胞的特征一致，诱导后的Rx+细胞共表达Pax6和有丝分裂标志物Ki67，但不表达Nestin。ES细胞来源的前体细胞在与胚胎视网膜细胞共培养时有效地产生具有光感受器表型(视紫红质(+)、恢复素(+))的细胞。此外，器官类型培养研究表明，ES细胞来源的细胞与视网膜外核层的光感受器表型(标记表达和形态)选择性整合和存活。综上所述，经SFEB/Dkk1/LeftyA/血清/激活素处理的ES细胞产生的视网膜神经前体细胞具有光感受器分化的能力。
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.