Stepwise differentiation of pluripotent stem cells into retinal cells
Stepwise differentiation of pluripotent stem cells into retinal cells
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DOI：
10.1038/nprot.2009.51
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发表时间：
2009-01-01
期刊：
影响因子：
14.8
通讯作者：
Takahashi, Masayo
中科院分区：
文献类型：
--
作者：
Osakada, Fumitaka;Ikeda, Hanako;Takahashi, Masayo
胚胎干细胞（ES细胞）是来源于囊胚期胚胎内细胞团的多能细胞。它们可以无限期地保持未分化状态，并且可以分化成所有三个胚层的衍生物，即外胚层、内胚层和中胚层。虽然在ES细胞的增殖和分化方面已经取得了很大的进展，但光感受器的诱导通常需要与发育中的视网膜组织共培养或移植到发育中的视网膜组织中。在这里，我们描述了一个协议，用于生成视网膜细胞从ES细胞逐步治疗与定义的因素。该方法优先从小鼠和人ES细胞诱导光感受器和视网膜色素上皮（RPE）细胞。在我们的方案中，从小鼠ES细胞分化RPE和光感受器需要28天，而人ES细胞分化为成熟的RPE和光感受器分别需要120和150天。这种分化系统和由此产生的多能干细胞衍生的视网膜细胞将促进视网膜疾病的移植疗法、药物测试和体外疾病建模的发展。它还将提高我们对中枢神经系统发育的理解，特别是眼睛。
Embryonic stem (ES) cells are pluripotent cells derived from the inner cell mass of blastocyst-stage embryos. They can maintain an undifferentiated state indefinitely and can differentiate into derivatives of all three germ layers, namely ectoderm, endoderm and mesoderm. Although much progress has been made in the propagation and differentiation of ES cells, induction of photoreceptors has generally required coculture with or transplantation into developing retinal tissue. Here, we describe a protocol for generating retinal cells from ES cells by stepwise treatment with defined factors. This method preferentially induces photoreceptor and retinal pigment epithelium (RPE) cells from mouse and human ES cells. In our protocol, differentiation of RPE and photoreceptors from mouse ES cells requires 28 d and the differentiation of human ES cells into mature RPE and photoreceptors requires 120 and 150 d, respectively. This differentiation system and the resulting pluripotent stem cell-derived retinal cells will facilitate the development of transplantation therapies for retinal diseases, drug testing and in vitro disease modeling. It will also improve our understanding of the development of the central nervous system, especially the eye.