Differentiation of primate ES cells into retinal cells induced by ES cell-derived pigmented cells.
Differentiation of primate ES cells into retinal cells induced by ES cell-derived pigmented cells.
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DOI：
10.1016/j.bbrc.2010.03.008
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发表时间：
2010-04
影响因子：
3.1
通讯作者：
Fengming Yue;K. Johkura;Sakiko Shirasawa;Tadayuki Yokoyama;Yuji Inoue;D. Tomotsune;K. Sasaki
中科院分区：
文献类型：
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作者：
Fengming Yue;K. Johkura;Sakiko Shirasawa;Tadayuki Yokoyama;Yuji Inoue;D. Tomotsune;K. Sasaki
目的：光感受器一旦紊乱，就不能再生和恢复其功能。视网膜色素上皮（retinal pigment epithelium，RPE）移植是近年来视网膜变性治疗的一种新方法。在本研究中，我们研究了通过共培养灵长类动物胚胎干细胞（ESCs）与ESCs衍生的RPE细胞的光感受器的诱导。方法：将胚胎干细胞与支持细胞共培养，获得RPE细胞。结果：RPE细胞的生成通过形态学分析证实，其显示具有紧密细胞-细胞排列的高度着色的多边形细胞。将胚胎干细胞和视网膜色素上皮细胞共培养后，一些胚胎干细胞衍生物对视紫红质呈免疫阳性。RT-PCR分析证实了视网膜相关基因标记物如Pax 6、CRX、IRBP、视紫红质、视紫红质激酶和Muschx 10A的表达。当加入RA时，光感受器特异性蛋白和基因的表达明显增加，此外，双极水平细胞的分化表现为蛋白和基因的表达。将与RPE细胞共培养并经RA处理的ESCs移植到裸鼠的肾包膜或玻璃体腔内。移植的ESC衍生物表现出广泛的视紫红质表达，它们存活并组织成受体组织，尽管它们形成畸胎瘤。结论：这些结果表明，与ESC衍生的RPE细胞共培养ESC是诱导光感受器的有用且有效的方法，并提供了对ESC用于视网膜再生的了解。
Purpose: Photoreceptors cannot regenerate and recover their functions once disordered. Transplantation of retinal pigment epithelium (RPE) has recently become a possible therapeutic approach for retinal degeneration. In the present study, we investigated the induction of photoreceptors by coculturing primate embryonic stem cells (ESCs) with ESC-derived RPE cells. Methods: RPE cells were derived by coculturing ESCs and Sertoli cells. Photoreceptors were then induced by using ESC-derived RPE cells and retinoic acid (RA) Results: RPE cell generation was confirmed by morphological analysis, which revealed highly pigmented polygonal cells with a compact cell–cell arrangement. After coculturing ESCs and RPE cells, some ESC derivatives became immunopositive for rhodopsin. RT-PCR analysis demonstrated the expression of retina-related gene markers such as Pax6, CRX, IRBP, rhodopsin, rhodopsin kinase, and Muschx10A. When RA was added, a distinct increase in the expression of photoreceptor-specific proteins and genes was found. In addition, the differentiation of bipolar horizontal cells was demonstrated by protein and gene expression. The ESCs that were cocultured with RPE cells and treated with RA were transplanted into the renal capsule or intra-vitreal space of nude mice. Grafted ESC derivatives demonstrated extensive rhodopsin expression, and they survived and organized into recipient tissues, although they formed teratomas. Conclusion: These results indicate that coculturing ESCs with ESC-derived RPE cells is a useful and efficient method for inducing photoreceptors and providing an insight into the use of ESCs for retina regeneration.