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Junji Hamuro, Kohsaku Numa, Tomoko Fujita, Munetoyo Toda, Koji Ueda, Yuichi Tokuda, Atushi Mukai, Masakazu Nakano, Morio Ueno, Shigeru Kinoshita, Chie Sotozono; Metabolites Interrogation in Cell Fate Decision of Cultured Human Corneal Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2020;61(2):10. doi: https://doi.org/10.1167/iovs.61.2.10.
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© ARVO (1962-2015); The Authors (2016-present)
Aiming to clarify the metabolic interrogation in cell fate decision of cultured human corneal endothelial cells (cHCECs).
To analyze the metabolites in the culture supernatants (CS), 34 metabolome measurements were carried out for mature differentiated and a variety of cHCECs with cell state transition through a facility service. Integrated proteomics research for cell lysates by liquid chromatography−tandem mass spectrometry (LC-MS/MS) was performed for 3 aliquots of each high-quality or low-quality cHCEC subpopulations (SP). The investigations for the focused genes involved in cHCEC metabolism were performed by using DAVID and its options “KEGG_PATHWAY.”
The clusters of metabolites coincided well with the distinct content of CD44−/+ SPs. Both secreted pyruvic acid and lactic acid in the CS were negatively correlated with the content of high-quality SPs. Lactic acid and pyruvic acid in the CS exhibited the positive correlation with that of Ile, Leu, and Ser, whereas the negative correlation was with glutamine. Platelet-derived growth factor-ββ in the CS negatively correlated with lactic acid in CS, indicating indirectly the positive correlation with the content of CD44−/+ SPs. Upregulated glycolytic enzymes and influx of glutamine to the tricarboxylic acid cycle may be linked with a metabolic rewiring converting oxidative metabolism in mature differentiated CD44−/+SPs into a glycolytic flux-dependent state in immature SPs with cell state transition.
The findings suggest that the cell fate decision of cHCECs may be dictated at least partly through metabolic rewiring.
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