Differentiation of RPE cells from integration-free iPS cells and their cell biological characterization.
Differentiation of RPE cells from integration-free iPS cells and their cell biological characterization.
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DOI：
10.1186/s13287-017-0652-9
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发表时间：
2017-10-02
影响因子：
7.5
通讯作者：
Williams DS
中科院分区：
文献类型：
--
作者：
Hazim RA;Karumbayaram S;Jiang M;Dimashkie A;Lopes VS;Li D;Burgess BL;Vijayaraj P;Alva-Ornelas JA;Zack JA;Kohn DB;Gomperts BN;Pyle AD;Lowry WE;Williams DS
视网膜色素上皮（RPE）的功能障碍涉及许多形式的视网膜变性。眼睛的易接近环境使其特别适合于RPE细胞的移植，RPE细胞现在可以来源于自体诱导多能干细胞（iPSC），以治疗视网膜变性。为了使RPE移植在临床上变得可行，患者特异性体细胞应该被重编程为iPSC，而不将重编程基因引入宿主细胞的基因组中，然后随后分化为在移植前充分表征安全性和功能性的RPE细胞。我们使用非整合RNA将人真皮成纤维细胞重编程为iPSC，并在良好生产规范（GMP）兼容的条件下将iPSC分化为RPE命运（iPSC-RPE）。使用高度敏感的细胞极性，结构，细胞器运输和功能的测定，我们发现培养中的iPSC-RPE细胞表现出天然RPE的关键特征。重要的是，我们第一次用任何干细胞衍生的RPE细胞证明活细胞能够支持动态细胞器运输。这种高度敏感的测试对于用于移植的RPE细胞至关重要，因为细胞内运动的缺陷已被证明促进RPE发病机制，类似于黄斑变性中发现的发病机制。为了测试它们的体内移植能力，我们将iPSC-RPE细胞注射到视网膜变性小鼠模型的视网膜下腔中，并证明移植的细胞能够挽救失去的RPE功能。本报告记录了在GMP相容条件下成功生成具有健康RPE特定特征的人iPSC-RPE细胞。该报告增加了越来越多的关于人类iPSC-RPE细胞用于致病性细胞培养研究和治疗性移植的文献，证实了其他人的发现，并提供了关于RPE细胞基本生物学特性的重要新信息。本文的在线版本（doi：10.1186/s13287-017-0652-9）包含补充材料，可供授权用户使用。
Dysfunction of the retinal pigment epithelium (RPE) is implicated in numerous forms of retinal degeneration. The readily accessible environment of the eye makes it particularly suitable for the transplantation of RPE cells, which can now be derived from autologous induced pluripotent stem cells (iPSCs), to treat retinal degeneration. For RPE transplantation to become feasible in the clinic, patient-specific somatic cells should be reprogrammed to iPSCs without the introduction of reprogramming genes into the genome of the host cell, and then subsequently differentiated into RPE cells that are well characterized for safety and functionality prior to transplantation. We have reprogrammed human dermal fibroblasts to iPSCs using nonintegrating RNA, and differentiated the iPSCs toward an RPE fate (iPSC-RPE), under Good Manufacturing Practice (GMP)-compatible conditions. Using highly sensitive assays for cell polarity, structure, organelle trafficking, and function, we found that iPSC-RPE cells in culture exhibited key characteristics of native RPE. Importantly, we demonstrate for the first time with any stem cell-derived RPE cell that live cells are able to support dynamic organelle transport. This highly sensitive test is critical for RPE cells intended for transplantation, since defects in intracellular motility have been shown to promote RPE pathogenesis akin to that found in macular degeneration. To test their capabilities for in-vivo transplantation, we injected the iPSC-RPE cells into the subretinal space of a mouse model of retinal degeneration, and demonstrated that the transplanted cells are capable of rescuing lost RPE function. This report documents the successful generation, under GMP-compatible conditions, of human iPSC-RPE cells that possess specific characteristics of healthy RPE. The report adds to a growing literature on the utility of human iPSC-RPE cells for cell culture investigations on pathogenicity and for therapeutic transplantation, by corroborating findings of others, and providing important new information on essential RPE cell biological properties. The online version of this article (doi:10.1186/s13287-017-0652-9) contains supplementary material, which is available to authorized users.
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DOI：
10.1073/pnas.1130432100
发表时间：
2003-05-27
影响因子：
11.1
作者：
Gibbs, D;Kitamoto, J;Williams, DS
通讯作者：
Williams, DS
影响因子：
4.8
作者：
AZARIAN, SM;KING, AJ;WILLIAMS, DS
通讯作者：
WILLIAMS, DS
DOI：
10.1083/jcb.109.6.2817
发表时间：
1989-12
期刊：
The Journal of cell biology
影响因子：
--
作者：
Bacallao R;Antony C;Dotti C;Karsenti E;Stelzer EH;Simons K
通讯作者：
Simons K
影响因子：
3.5
作者：
D'Cruz, PM;Yasumura, D;Vollrath, D
通讯作者：
Vollrath, D
影响因子：
17.8
作者：
Anderson, Don H.;Radeke, Monte J.;Gallo, Natasha B.;Chapin, Ethan A.;Johnson, Patrick T.;Curletti, Christy R.;Hancox, Lisa S.;Hu, Jane;Ebright, Jessica N.;Malek, Goldis;Hauser, Michael A.;Rickman, Catherine Bowes;Bok, Dean;Hageman, Gregory S.;Johnson, Lincoln V.
通讯作者：
Johnson, Lincoln V.