Toward the generation of rod and cone photoreceptors from mouse, monkey and human embryonic stem cells
Toward the generation of rod and cone photoreceptors from mouse, monkey and human embryonic stem cells
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DOI：
10.1038/nbt1384
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发表时间：
2008-02-01
影响因子：
46.9
通讯作者：
Takahashi, Masayo
中科院分区：
文献类型：
--
作者：
Osakada, Fumitaka;Ikeda, Hanako;Takahashi, Masayo
我们之前报道了小鼠胚胎干细胞（ES）分化为视网膜祖细胞。然而，这些祖细胞很少分化成光感受器，除非它们与胚胎视网膜组织一起培养。在这里，我们展示了在没有视网膜组织的情况下，通过在规定的培养条件下逐步处理，从小鼠、猴子和人类胚胎干细胞中体外产生假定的杆状和锥状光感受器。用Notch信号抑制剂处理小鼠胚胎干细胞，从Rx（+）视网膜祖细胞诱导出Crx（+）光受体前体。进一步应用成纤维细胞生长因子、Shh、牛磺酸和视黄酸，除了产生默认的视锥外，还产生了更多的视紫红质（+）杆状光感受器。用猴和人胚胎干细胞，联合Wnt和Nodal抑制剂进行无饲料和无血清悬浮培养，诱导Rx（+）或Mitf（+）视网膜祖细胞分化，产生视网膜色素上皮细胞。随后用视黄酸和牛磺酸诱导光感受器分化。这些发现可能促进基于人类胚胎干细胞移植治疗视网膜疾病的发展。
We previously reported the differentiation of mouse embryonic stem (ES) cells into retinal progenitors. However, these progenitors rarely differentiate into photoreceptors unless they are cultured with embryonic retinal tissues. Here we show the in vitro generation of putative rod and cone photoreceptors from mouse, monkey and human ES cells by stepwise treatments under defined culture conditions, in the absence of retinal tissues. With mouse ES cells, Crx(+) photoreceptor precursors were induced from Rx(+) retinal progenitors by treatment with a Notch signal inhibitor. Further application of fibroblast growth factors, Shh, taurine and retinoic acid yielded a greater number of rhodopsin(+) rod photoreceptors, in addition to default cone production. With monkey and human ES cells, feeder- and serum-free suspension culture combined with Wnt and Nodal inhibitors induced differentiation of Rx(+) or Mitf(+) retinal progenitors, which produced retinal pigment epithelial cells. Subsequent treatment with retinoic acid and taurine induced photoreceptor differentiation. These findings may facilitate the development of human ES cell-based transplantation therapies for retinal diseases.