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人胚胎干细胞来源与原代人视网膜色素上皮的比较蛋白质组学分析。

Comparative proteomic analysis of human embryonic stem cell-derived and primary human retinal pigment epithelium.

机构信息

BioMediTech Institute, Faculty of Medicine and Life Sciences, University of Tampere, Tampere, Finland.

Department of Ophthalmology, Faculty of Medicine and Life Sciences, University of Tampere, Tampere, Finland.

出版信息

Sci Rep. 2017 Jul 20;7(1):6016. doi: 10.1038/s41598-017-06233-9.

Abstract

Human embryonic stem cell-derived retinal pigment epithelial cells (hESC-RPE) provide an unlimited cell source for retinal cell replacement therapies. Clinical trials using hESC-RPE to treat diseases such as age-related macular degeneration (AMD) are currently underway. Human ESC-RPE cells have been thoroughly characterized at the gene level but their protein expression profile has not been studied at larger scale. In this study, proteomic analysis was used to compare hESC-RPE cells differentiated from two independent hESC lines, to primary human RPE (hRPE) using Isobaric tags for relative quantitation (iTRAQ). 1041 common proteins were present in both hESC-RPE cells and native hRPE with majority of the proteins similarly regulated. The hESC-RPE proteome reflected that of normal hRPE with a large number of metabolic, mitochondrial, cytoskeletal, and transport proteins expressed. No signs of increased stress, apoptosis, immune response, proliferation, or retinal degeneration related changes were noted in hESC-RPE, while important RPE specific proteins involved in key RPE functions such as visual cycle and phagocytosis, could be detected in the hESC-RPE. Overall, the results indicated that the proteome of the hESC-RPE cells closely resembled that of their native counterparts.

摘要

人胚胎干细胞来源的视网膜色素上皮细胞(hESC-RPE)为视网膜细胞替代疗法提供了无限的细胞来源。目前正在进行使用 hESC-RPE 治疗年龄相关性黄斑变性(AMD)等疾病的临床试验。人类 ESC-RPE 细胞在基因水平上已经得到了彻底的描述,但它们的蛋白质表达谱尚未在更大的范围内进行研究。在这项研究中,使用等相对和绝对定量标记(iTRAQ)对来自两个独立的 hESC 系的 hESC-RPE 细胞与原代人 RPE(hRPE)进行了蛋白质组学分析比较。在 hESC-RPE 细胞和天然 hRPE 中存在 1041 种共同蛋白质,大多数蛋白质的调节方式相似。hESC-RPE 蛋白组反映了正常 hRPE 的情况,大量表达代谢、线粒体、细胞骨架和转运蛋白。在 hESC-RPE 中未观察到应激、凋亡、免疫反应、增殖或与视网膜变性相关的变化迹象,而在 hESC-RPE 中可以检测到参与关键 RPE 功能(如视觉循环和吞噬作用)的重要 RPE 特异性蛋白质。总体而言,结果表明 hESC-RPE 细胞的蛋白质组与其天然对应物非常相似。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a6d4/5519552/890411b9a6fb/41598_2017_6233_Fig1_HTML.jpg

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