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

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作者
Kassandra Groux
Anna Verschueren
Céline Nanteau
Marilou Clémençon
Mathias Fink
José-Alain Sahel
Claude Boccara
Michel Paques
Sacha Reichman
Kate Grieve
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[1] CNRS,Institut Langevin, ESPCI Paris
[2] CIC 1423,PSL
[3] CNRS,Paris Eye Imaging Group, Quinze
[4] Fondation Ophtalmologique Adolphe de Rotschild,Vingts National Eye Hospital, INSERM
[5] The University of Pittsburgh School of Medicine,DGOS
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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.
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