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

• Published in: Communications biology Vol. 5; no. 1; p. 575
• Main Authors: Groux, Kassandra, Verschueren, Anna, Nanteau, Céline, Clémençon, Marilou, Fink, Mathias, Sahel, José-Alain, Boccara, Claude, Paques, Michel, Reichman, Sacha, Grieve, Kate
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 10.06.2022; Nature Publishing Group; Nature Portfolio
• Subjects: 631/1647/245/2226; 631/57; Age; Animals; Atrophy; Biology; Biomedical and Life Sciences; Cellular Biology; Cytoskeleton; Degenerative diseases; Dystrophy; Epithelium; Fluorescein Angiography; Humans; Immunohistochemistry; Life Sciences; Macular degeneration; Macular Degeneration - metabolism; Mitochondria; Organelles; Phenotypes; Pluripotency; Retina; Retinal degeneration; Retinal pigment epithelium; Retinal Pigment Epithelium - metabolism; Stem cells; Subcellular Processes; Swine; Tomography; Tomography, Optical Coherence - methods; Wound healing
• ISSN: 2399-3642; 2399-3642
• DOI: 10.1038/s42003-022-03479-6

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. Dynamic full-field optical coherence tomography (D-FFOCT) is used for live cell imaging of primary porcine retinal pigment epithelium (ppRPE) cultures and human induced pluripotent stem cell-derived RPE (hiRPE) cultures, allowing non-invasive realtime access to organelles and cytoskeleton dynamics in RPE cells.