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动态全场光学相干断层扫描可实时成像视网膜色素上皮应激模型。

Dynamic full-field optical coherence tomography allows live imaging of retinal pigment epithelium stress model.

机构信息

Institut Langevin, ESPCI Paris-PSL, CNRS, 10 rue Vauquelin, Paris, 75005, France.

Paris Eye Imaging Group, Quinze-Vingts National Eye Hospital, INSERM-DGOS, CIC 1423, 28 rue de Charenton, Paris, 75012, France.

出版信息

Commun Biol. 2022 Jun 10;5(1):575. doi: 10.1038/s42003-022-03479-6.

Abstract

Retinal degenerative diseases lead to the blindness of millions of people around the world. In case of age-related macular degeneration (AMD), the atrophy of retinal pigment epithelium (RPE) precedes neural dystrophy. But as crucial as understanding both healthy and pathological RPE cell physiology is for those diseases, no current technique allows subcellular in vivo or in vitro live observation of this critical cell layer. To fill this gap, we propose dynamic full-field OCT (D-FFOCT) as a candidate for live observation of in vitro RPE phenotype. In this way, we monitored primary porcine and human stem cell-derived RPE cells in stress model conditions by performing scratch assays. In this study, we quantified wound healing parameters on the stressed RPE, and observed different cell phenotypes, displayed by the D-FFOCT signal. In order to decipher the subcellular contributions to these dynamic profiles, we performed immunohistochemistry to identify which organelles generate the signal and found mitochondria to be the main contributor to D-FFOCT contrast. Altogether, D-FFOCT appears to be an innovative method to follow degenerative disease evolution and could be an appreciated method in the future for live patient diagnostics and to direct treatment choice.

摘要

视网膜退行性疾病导致全世界数百万人失明。在年龄相关性黄斑变性 (AMD) 的情况下,视网膜色素上皮 (RPE) 的萎缩先于神经营养不良。但是,了解健康和病理 RPE 细胞生理学对于这些疾病至关重要,目前没有技术可以对这一关键细胞层进行亚细胞的体内或体外活观察。为了填补这一空白,我们提出动态全场 OCT (D-FFOCT) 作为体外 RPE 表型活体观察的候选方法。通过划痕实验,我们在应激模型条件下监测了原代猪和人干细胞衍生的 RPE 细胞。在这项研究中,我们量化了应激 RPE 的伤口愈合参数,并通过 D-FFOCT 信号观察到不同的细胞表型。为了解码这些动态图谱的亚细胞贡献,我们进行了免疫组织化学分析以确定哪些细胞器产生信号,并发现线粒体是 D-FFOCT 对比的主要贡献者。总之,D-FFOCT 似乎是一种用于跟踪退行性疾病进展的创新方法,并且在未来可能成为一种有价值的活体患者诊断方法,并指导治疗选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e80f/9187748/8337552dcec9/42003_2022_3479_Fig1_HTML.jpg

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