Hypoxia Increases the Yield of Photoreceptors Differentiating from Mouse Embryonic Stem Cells and Improves the Modeling of Retinogenesis In Vitro
Hypoxia Increases the Yield of Photoreceptors Differentiating from Mouse Embryonic Stem Cells and Improves the Modeling of Retinogenesis In Vitro
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DOI：
10.1002/stem.1339
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发表时间：
2013-05-01
期刊：
影响因子：
5.2
通讯作者：
Bhattacharya, Shomi S.
中科院分区：
文献类型：
--
作者：
Garita-Hernandez, Marcela;Diaz-Corrales, Francisco;Bhattacharya, Shomi S.
色素性视网膜炎（RP）是一种遗传异质性疾病，与年龄相关性黄斑变性（AMD）一起，是导致永久性失明的主要原因，其特征是视网膜感光细胞的进行性功能障碍和死亡。由于哺乳动物视网膜的再生能力有限，科学界投入了大量资金，试图从胚胎干细胞（ESC）中获得视网膜祖细胞。这代表了视网膜细胞的无限来源，但目前还不可能达到特定的群体，如光感受器，有效地使它们能够在未来安全地用于RP或AMD的细胞治疗。在这项研究中，我们通过再现视网膜发育的关键阶段，从小鼠ESC （mESC）的定向分化中获得了高产量的光感受器。我们提出了一种新的分化方案，包括缺氧，并考虑到外在和内在的线索。这些包括特定的小生境条件以及视网膜发育过程中涉及的信号通路的操纵。我们的研究结果表明，缺氧促进和改善mESC向光感受器的分化。免疫荧光分析显示，在低氧条件下，不同群体的视网膜细胞数量增加，如crx阳性细胞，s - opsin阳性细胞，Rhodopsin和Recoverin双阳性细胞。本文首次报道了在野生型小鼠视网膜下空间移植的大量分化细胞在体内的高效分化和成熟的杆状光感受器标记物的表达。干细胞2013；31:966 - 978
Retinitis pigmentosa (RP), a genetically heterogeneous group of diseases together with age-related macular degeneration (AMD), are the leading causes of permanent blindness and are characterized by the progressive dysfunction and death of the light sensing photoreceptors of the retina. Due to the limited regeneration capacity of the mammalian retina, the scientific community has invested significantly in trying to obtain retinal progenitor cells from embryonic stem cells (ESC). These represent an unlimited source of retinal cells, but it has not yet been possible to achieve specific populations, such as photoreceptors, efficiently enough to allow them to be used safely in the future as cell therapy of RP or AMD. In this study, we generated a high yield of photoreceptors from directed differentiation of mouse ESC (mESC) by recapitulating crucial phases of retinal development. We present a new protocol of differentiation, involving hypoxia and taking into account extrinsic and intrinsic cues. These include niche-specific conditions as well as the manipulation of the signaling pathways involved in retinal development. Our results show that hypoxia promotes and improves the differentiation of mESC toward photoreceptors. Different populations of retinal cells are increased in number under the hypoxic conditions applied, such as Crx-positive cells, S-Opsin-positive cells, and double positive cells for Rhodopsin and Recoverin, as shown by immunofluorescence analysis. For the first time, this manuscript reports the high efficiency of differentiation in vivo and the expression of mature rod photoreceptor markers in a large number of differentiated cells, transplanted in the subretinal space of wild-type mice. STEM CELLS 2013;31:966-978