High-Throughput Microscopy Analysis of Mitochondrial Membrane Potential in 2D and 3D Models

Recent proteomic, metabolomic, and transcriptomic studies have highlighted a connection between changes in mitochondria physiology and cellular pathophysiological mechanisms. Secondary assays to assess the function of these organelles appear fundamental to validate these -omics findings. Although mi...

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Vydáno v:	Cells (Basel, Switzerland) Ročník 12; číslo 7; s. 1089
Hlavní autoři:	Vianello, Caterina, Dal Bello, Federica, Shin, Sang Hun, Schiavon, Sara, Bean, Camilla, Magalhães Rebelo, Ana Paula, Knedlík, Tomáš, Esfahani, Emad Norouzi, Costiniti, Veronica, Lacruz, Rodrigo S., Covello, Giuseppina, Munari, Fabio, Scolaro, Tommaso, Viola, Antonella, Rampazzo, Elena, Persano, Luca, Zumerle, Sara, Scorrano, Luca, Gianelle, Alessio, Giacomello, Marta
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	Switzerland MDPI AG 01.04.2023 MDPI
Témata:	Amino acids Analysis Blood & organ donations Care and treatment Cell culture co-culture Diagnosis Dosage and administration Dyes Fibroblasts Fibroblasts - metabolism Health aspects High-throughput screening (Biochemical assaying) Homeostasis Humans Image processing Kinetics Macrophages Melanoma Membrane potential Membrane Potential, Mitochondrial Metabolomics Microscopy Mitochondria Mitochondria - metabolism Mitochondrial diseases mitochondrial membrane potential Neural stem cells NSCs Organelles Proteomics Respiration single muscle fibers Spheroids Tetracycline Tetracyclines TMRM Transcriptomics Italy single muscle fibers NSCs TMRM mitochondrial membrane potential machine learning spheroids co-culture
ISSN:	2073-4409, 2073-4409
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Shrnutí:	Recent proteomic, metabolomic, and transcriptomic studies have highlighted a connection between changes in mitochondria physiology and cellular pathophysiological mechanisms. Secondary assays to assess the function of these organelles appear fundamental to validate these -omics findings. Although mitochondrial membrane potential is widely recognized as an indicator of mitochondrial activity, high-content imaging-based approaches coupled to multiparametric to measure it have not been established yet. In this paper, we describe a methodology for the unbiased high-throughput quantification of mitochondrial membrane potential in vitro, which is suitable for 2D to 3D models. We successfully used our method to analyze mitochondrial membrane potential in monolayers of human fibroblasts, neural stem cells, spheroids, and isolated muscle fibers. Moreover, by combining automated image analysis and machine learning, we were able to discriminate melanoma cells from macrophages in co-culture and to analyze the subpopulations separately. Our data demonstrated that our method is a widely applicable strategy for large-scale profiling of mitochondrial activity.
Bibliografie:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 Current address: Department of Biochemistry and Molecular Pharmacology, New York University Medical Center, New York, NY 10016, USA. These authors contributed equally to this work.
ISSN:	2073-4409 2073-4409
DOI:	10.3390/cells12071089