Generation of retinal pigment epithelial cells from human embryonic stem cell-derived spherical neural masses
Generation of retinal pigment epithelial cells from human embryonic stem cell-derived spherical neural masses
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DOI：
10.1016/j.scr.2012.05.002
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发表时间：
2012-09-01
影响因子：
1.2
通讯作者：
Yu, Hyeong Gon
中科院分区：
文献类型：
--
作者：
Cho, Myung Soo;Kim, Sang Jin;Yu, Hyeong Gon
视网膜色素上皮（retinal pigment epithelium，RPE）的功能障碍和丧失是在各种视网膜退行性疾病如老年性黄斑变性中观察到的主要病理变化。从人多能干细胞产生的RPE可以是RPE替代疗法的良好候选者。在这里，我们显示了人胚胎干细胞（hESC）向RPE的分化，产生球形神经块（SNMs），这是纯群众的hESC衍生的神经前体。在SNMs的早期传代过程中，从开放的神经管样结构产生的囊性结构显示出色素上皮形态。这些色素细胞通过神经外胚层诱导和机械纯化分化为功能性RPE。大多数分化的细胞表现出典型的RPE形态，如杯状上皮单层，透射电子显微镜显示顶端微绒毛，色素颗粒，和紧密连接。这些细胞还表达RPE的分子标记物，包括Mitf、ZO-1、RPE 65、CRALBP和雌激素样蛋白。所产生的RPE还显示出对分离的牛光感受器外节的吞噬作用，并分泌色素上皮衍生因子和血管内皮生长因子。在我们的方法中，可以从SNM生成功能性RPE。由于SNM具有几个优点，包括长期扩增而不丧失分化能力的能力，易于储存和解冻，并且不需要饲养细胞，因此我们的RPE分化方法可以用作产生用于视网膜再生治疗的功能性RPE细胞的有效策略。(c)2012爱思唯尔有限公司版权所有。
Dysfunction and loss of retinal pigment epithelium (RPE) are major pathologic changes observed in various retinal degenerative diseases such as aged-related macular degeneration. RPE generated from human pluripotent stem cells can be a good candidate for RPE replacement therapy. Here, we show the differentiation of human embryonic stem cells (hESCs) toward RPE with the generation of spherical neural masses (SNMs), which are pure masses of hESCs-derived neural precursors. During the early passaging of SNMs, cystic structures arising from opened neural tube-like structures showed pigmented epithelial morphology. These pigmented cells were differentiated into functional RPE by neuroectodermal induction and mechanical purification. Most of the differentiated cells showed typical RPE morphologies, such as a polygonal-shaped epithelial monolayer, and transmission electron microscopy revealed apical microvilli, pigment granules, and tight junctions. These cells also expressed molecular markers of RPE, including Mitf, ZO-1, RPE65, CRALBP, and bestrophin. The generated RPE also showed phagocytosis of isolated bovine photoreceptor outer segment and secreting pigment epithelium-derived factor and vascular endothelial growth factor. Functional RPE could be generated from SNM in our method. Because SNMs have several advantages, including the capability of expansion for long periods without loss of differentiation capability, easy storage and thawing, and no need for feeder cells, our method for RPE differentiation may be used as an efficient strategy for generating functional RPE cells for retinal regeneration therapy. (c) 2012 Elsevier B.V. All rights reserved.